Multiomics characterization of cell type repertoires for urine liquid biopsies.

Urine is assayed alongside blood in medicine, yet current clinical diagnostic tests utilize only a small fraction of its total biomolecular repertoire, potentially foregoing high-resolution insights into human health and disease. In this work, we characterized the joint landscapes of transcriptomic and metabolomic signals in human urine. We also compared the urine transcriptome to plasma cell-free RNA, identifying a distinct cell type repertoire and enrichment for metabolic signal. Untargeted metabolomic measurements identified a complementary set of pathways to the transcriptomic analysis. Our findings suggest that urine is a promising biofluid yielding prognostic and detailed insights for hard-to-biopsy tissues with low representation in the blood, offering promise for a new generation of liquid biopsies.

The Omega-3 Index Response to an 8 Week Randomized Intervention Containing Three Fatty Fish Meals Per Week Is Influenced by Adiposity in Overweight to Obese Women.

BACKGROUND: The Dietary Guidelines for Americans (DGA) recommends consuming ~225 g/wk of a variety of seafood providing >1.75 g/wk of long-chain omega-3 fatty acids to reduce cardiovascular disease risk, however individual responses to treatment vary. OBJECTIVE: This study had three main objectives. First, to determine if a DGA-conforming diet (DGAD), in comparison to a typical American diet (TAD), can increase the omega-3 index (OM3I), i.e., the red blood cell mol% of eicosapentaenoic acid (EPA) + docosahexaenoic acid (DHA). Second, to identify factors explaining variability in the OM3I response to dietary treatment. Third to identify factors associated with the baseline OM3I. DESIGN: This is a secondary analysis of a randomized, double-blind 8 wk dietary intervention of overweight/obese women fed an 8d rotating TAD (n = 20) or DGAD (n = 22) registered at www.clinicaltrials.gov as NCT02298725. The DGAD-group consumed 240 g/wk of Atlantic farmed salmon and albacore tuna in three meals with an estimated EPA + DHA of 3.7 ± 0.6 g/wk. The TAD-group consumed ~160 g/wk of farmed white shrimp and a seafood salad containing imitation crab in three meal with an estimated EPA + DHA of 0.45 ± 0.05 g/wk. Habitual diet was determined at baseline, and body composition was determined at 0 and 8wks. Red blood cell fatty acids were measured at 0, 2 and 8 wk. RESULTS: At 8 wk, the TAD-group OM3I was unchanged (5.90 ± 1.35-5.80 ± 0.76%), while the DGAD-group OM3I increased (5.63 ± 1.27-7.33 ± 1.36%; p < 0.001). In the DGAD-group 9 of 22 participants achieved an OM3I >8%. Together, body composition and the baseline OM3I explained 83% of the response to treatment variability. Baseline OM3I (5.8 ± 1.3%; n = 42) was negatively correlated to the android fat mass (p = 0.0007) and positively correlated to the FFQ estimated habitual (EPA+DHA) when expressed as a ratio to total dietary fat (p = 0.006). CONCLUSIONS: An 8 wk TAD did not change the OM3I of ~6%, while a DGAD with 240 g/wk of salmon and albacore tuna increased the OM3I. Body fat distribution and basal omega-3 status are primary factors influencing the OM3I response to dietary intake in overweight/obese women.

Plasma and serum oxylipin, endocannabinoid, bile acid, steroid, fatty acid and nonsteroidal anti-inflammatory drug quantification in a 96-well plate format.

The goal of this research was to develop a high-throughput, cost-effective method for metabolic profiling of lipid mediators and hormones involved in the regulation of inflammation and energy metabolism, along with polyunsaturated fatty acids and common over-the-counter non-steroidal anti-inflammatory drugs (NSAIDs). We describe a 96-well plate protein precipitation and filtration procedure for 50 µL of plasma or serum in the presence of 37 deuterated analogs and 2 instrument internal standards. Data is acquired in two back-to-back UPLC-MS/MS analyses using electrospray ionization with positive/negative switching and scheduled multiple reaction monitoring for the determination of 145 compounds, including oxylipins, endocannabinoids and like compounds, bile acids, glucocorticoids, sex steroids, polyunsaturated fatty acids, and 3 NSAIDs. Intra- and inter-batch variability was <25% for >70% of metabolites above the LOQ in both matrices, but higher inter-batch variability was observed for serum oxylipins and some bile acids. Results for NIST Standard Reference Material 1950, compared favorably with the 20 certified metabolite values covered by this assay, and we provide new data for oxylipins, N-acylethanolamides, glucocorticoids, and 17-hydroxy-progesterone in this material. Application to two independent cohorts of elderly men and women showed the routine detection of 86 metabolites, identified fasting state influences on essential fatty acid-derived oxylipins, N-acylethanolamides and conjugated bile acids, identified rare presence of high and low testosterone levels and the presence of NSAIDs in ∼10% of these populations. The described method appears valuable for investigations in large cohort studies to provide insight into metabolic cross-talk between the array of mediators assessed here.

Associations Among Fatty Acids, Desaturase and Elongase, and Insulin Resistance in Children.

OBJECTIVES: Fatty acid profiles and desaturase (SCD-16, SCD018, D5D, D6D) and elongase (ELOVL6) enzyme activity have been associated with adiposity and metabolic disease. While this has been studied in adults, few studies have included children. The objective of this study was to evaluate these markers in children and identify relationships with markers of metabolic health. It was hypothesized that these lipid markers would be correlated to adiposity and metabolic disease. METHODS: This study was a cross-sectional analysis of fourth- and fifth-grade children (n = 86, aged 9-12) participating in a comprehensive nutrition program. Any student enrolled in the program was eligible for inclusion in this study. Fasting plasma was collected and analyzed for total fatty acids, glucose, insulin, and full lipid panels. Insulin resistance was estimated using calculated homeostatic model assessment for insulin resistance (HOMA-IR) values. RESULTS: There were no differences in lipid markers, glucose, insulin, or HOMA-IR among children classified as normal weight, overweight, or obese. SCD-16, D5D, and ELOVL6 activity was significantly correlated to HOMA-IR values (r = 0.39, p = 0.001; r = -0.33, p = 0.006; r = -0.37, p = 0.005, respectively). In regression analysis, body mass index for age percentile, D6D activity, ELOVL6 activity, and systolic blood pressure were the most significant predictors of HOMA-IR values (adjusted r2 = 0.39, p ≤ 0.001). CONCLUSIONS: There was no relationship between these lipid markers and adiposity in this population; however, there were correlations with HOMA-IR. Regardless of adiposity, there may be underlying changes in fatty acid and lipid metabolism associated with the development of metabolic diseases.