Single Blood Sample Research Articles

PSIV-22 Validation of 26a and let7 microRNA as early pregnancy markers in single cow blood samples, at day 28 after embryo transfer

Abstract In this work we validated the previous scientific reports done in pooled samples by two different groups, over the miRNA expression in single blood from pregnant cows. Particularly we indagate the expression at day 28 after embryo transfer of let 7a, and 26a as well as 16 microRNAs (miRNA) family. By qPCR technique, we demonstrated that let 7a, and 26a increase maximum 2.1 and 2.6 fold change respectively in pregnant vs non-pregnant nulliparous heifers. This represents an exciting opportunity to use the differential expression of these miRNAs as early pregnancy markers and better understand the relationship with the cellular and molecular interaction between mother and embryo.

Assessing maternal clotting function with novel global coagulation assays: A prospective pilot study.

Women are at higher risk of venous thromboembolism (VTE) during pregnancy and the puerperium. Global coagulation assays (GCAs), including thromboelastography (TEG), thrombin generation using the calibrated automated thrombogram (CAT) and fibrin generation using the overall haemostatic potential assay (OHP), provide a more comprehensive assessment of the coagulation process than conventional coagulation assays. We aimed to evaluate the ability of these GCAs to analyse the coagulability among pregnant women of varying VTE risk profile. Women undergoing term elective caesarean delivery provided a single predelivery blood sample for conventional and novel coagulation testing (TEG, CAT and OHP). Data from 47 healthy nonpregnant women aged 18-45years were used as controls. Sixty women with term singleton pregnancies were included. Samples from pregnant women were hypercoagulable on most GCA parameters compared to nonpregnant controls, demonstrating increased maximum amplitude (clot strength) (71.5 vs 60.6mm, P<.001) on whole blood TEG and increased endogenous thrombin potential (1895.22 vs 1399.33nmol/L·min, P<.001) and overall coagulation potential (fibrin generation) (57.58 vs 36.21 units, P<.001) on platelet-poor plasma. Pregnant women with booking BMI≥30kg/m2 had significantly higher maximum amplitude compared to pregnant women of normal BMI (18.5-25kg/m2 ) (73.2 vs 66.1mm, P<.001). Global coagulation assays reliably detect the physiological hypercoagulability of pregnancy. Thromboelastography in particular appears to correlate with obesity in the pregnant population. GCAs may be potential adjuncts to risk factor-based criteria to guide VTE thromboprophylaxis during pregnancy and the puerperium.

Transcutaneous CO2 Monitoring in Children Undergoing Tonsillectomy for Sleep Disordered Breathing.

Children undergoing tonsillectomy for sleep-disordered breathing are at risk for respiratory compromise when narcotics are administered. Severe complications resulting from hypoxia can include neuro-devastation and death. The objective of this prospective study was to evaluate the feasibility, accuracy, and utility of transcutaneous carbon dioxide (tcPCO2) monitoring during and after adenotonsillectomy. Prospective, Observational study. Twenty-nine children with sleep-disordered breathing scheduled for adenotonsillectomy were included in the study. Peri-operative measurements of tcPCO2 were compared against a single venous blood sample (PaCO2) and end-tidal (ET) CO2. The differences between ETCO2, tcPCO2 measures, and PaCO2 were examined using non-paired t-tests and linear regression. Parameters from PSG were recorded and associations with tcPCO2 values were performed using linear regression analysis. Group comparisons were made between pre-, intra-, and post-operative tcPCO2 measurements. Similar to ETCO2, there was good correlation of tcPCO2 to PaCO2. Children with lower oxygen (O2) saturation nadirs had higher peak CO2 levels during surgery and spent a greater proportion of time with CO2 > 50 mmHg in the recovery room (P < .01 and P < .08). Other PSG measures (apnea-hypopnea index, O2 desaturation index, and peak CO2) did not have any significant correlation. Frequent episodes of hypercapnia were noted intra- and post-operatively and mean tcPCO2 values during both periods were significantly higher than baseline (P < .001). tcPCO2 monitoring is viable in children undergoing adenotonsillectomy and can provide a good estimate of hypoventilation. tcPCO2 measurements may have particular benefit in the post-operative setting and may assist in identifying children at greater risk for respiratory complications. 4 Laryngoscope, 131:1410-1415, 2021.

Blood Parasites in Endangered Wildlife-Trypanosomes Discovered During a Survey of Haemoprotozoa from the Tasmanian Devil.

The impact of emerging infectious diseases is increasingly recognised as a major threat to wildlife. Wild populations of the endangered Tasmanian devil, Sarcophilus harrisii, are experiencing devastating losses from a novel transmissible cancer, devil facial tumour disease (DFTD); however, despite the rapid decline of this species, there is currently no information on the presence of haemoprotozoan parasites. In the present study, 95 Tasmanian devil blood samples were collected from four populations in Tasmania, Australia, which underwent molecular screening to detect four major groups of haemoprotozoa: (i) trypanosomes, (ii) piroplasms, (iii) Hepatozoon, and (iv) haemosporidia. Sequence results revealed Trypanosoma infections in 32/95 individuals. Trypanosoma copemani was identified in 10 Tasmanian devils from three sites and a second Trypanosoma sp. was identified in 22 individuals that were grouped within the poorly described T. cyclops clade. A single blood sample was positive for Babesia sp., which most closely matched Babesia lohae. No other blood protozoan parasite DNA was detected. This study provides the first insight into haemoprotozoa from the Tasmanian devil and the first identification of Trypanosoma and Babesia in this carnivorous marsupial.

A prospective study of placental growth factor in twin pregnancy and development of a dichorionic twin pregnancy specific reference range.

To develop a dichorionic twin pregnancy specific reference range for placental growth factor (PlGF), and to compare gestation-specific placental growth factor levels in twin pregnancies later complicated by pre-eclampsia, hypertensive disorder of pregnancy or fetal growth restriction with control pregnancies. Prospective observational study. Single large tertiary maternity unit in Ireland. Women with a twin pregnancy. Consenting pregnant women, across a variety of gestations, had a single blood sample taken at one time-point only during their pregnancy. The plasma was initially biobanked and PlGF was measured later in batches using the point of care Triage® PlGF test. Development of pre-eclampsia, hypertensive disorder of pregnancy or fetal growth restriction. Placental growth factor levels in uncomplicated dichorionic twin pregnancies were significantly lower in the women who later developed pre-eclampsia than in the controls at all gestational intervals. In those that later developed any hypertensive disorder of pregnancy, median PlGF was lower only in those recruited before 24weeks of gestation, whereas in infants with a customised birthweight below the third centile, PlGF was lower only in those sampled after 24weeks of gestation. Placental growth factor levels in twin pregnancy differ significantly between those women with a pregnancy that will later be complicated by pre-eclampsia and those that will not. This difference is present many weeks before clinical signs or symptoms of disease are present. Using cross-sectional values from uncomplicated twin pregnancies, we have developed a dichorionic twin pregnancy specific reference range for PlGF. Placental growth factor levels in twin pregnancy differ significantly between women that will later develop pre-eclampsia and those that will not.

New gender-specific formulae for estimating extracellular fluid volume from height and weight in adults.

First, to derive gender-specific formulae for estimation of extracellular fluid volume (eECV) and second, compare eECV as a scaling metric for slope-intercept glomerular filtration rate (GFR) with estimated body surface area (eBSA), lean body mass (eLBM) and total body water (eTBW). GFR and 'slope-only' GFR (GFR/ECV), both single compartment-corrected, were measured in a previously published multicentre database of healthy potential kidney transplant donors. Measured ECV (mECV) was obtained as ratio GFR-to-GFR/ECV. Formulae for eECV in men and women were derived from the relationship of mECV with height and weight and expressed as eECV = a.weight.height. In a population of prospective kidney transplant donors from a single centre, eECV was compared with mECV. GFR was scaled to eECV, eBSA, eLBM and eTBW, estimated from previously published formulae. In men and women, respectively, a was 0.0755 and 0.0399, x was 0.6185 and 0.6065 and y was 0.4982 and 0.6217. In the single centre, biases (±precisions) of eECV against mECV in men and women were 0.26 (±1.68) and 0.31 (±1.67) l. Mean GFR/eBSA was higher in men but mean GFR/eLBM and GFR/eTBW were higher in women. Mean GFR/ECV and mean GFR/eECV were very similar between the two genders. GFR/ECV and GFR/eECV showed correlations with each other that were almost identical between men and women. New formulae are described for estimating eECV. Scaling GFR to eECV is more physiological than scaling to eBSA and accounts for gender. eECV used for measuring GFR from a single blood sample should be gender-specific.

Variation of HLA class I (-A and -C) genes in individuals infected with hepatitis B or hepatitis C virus in Cameroon

The Human Leucocyte Antigens (HLA) work in concert with other immune factors to modulate immunity to viral infections. Extensive variation has been reported in the genetic sequences and functions of classical HLA class I genes in many (mostly Western) populations, and several HLA associations with infectious disease outcomes have been reported. Little is known about their role in the susceptibility or resistance to hepatitis viruses in Central African populations. The aim of this study was to determine variants of two HLA class I genes (HLA-A and -C) in adults infected with hepatitis B (HBV)- or -C (HCV) virus in Cameroon.In this case-control study, a total of 169 unrelated adults comprising 68 HCV-infected, 38 HBV-infected and 63 uninfected (controls) individuals participated. Each consented participant was screened for HBV, HCV, and HIV infections and willingly donated a single blood sample for genomic DNA isolation and some clinical laboratory tests. HLA-A and HLA-C were genotyped using previously described sequence-based techniques (SBT).A total of 54 HLA alleles were identified in the study population (27 HLA-A and 27 HLA-C). HLA-A∗23:01 and HLA-C∗07:01 were the most common alleles with genotype frequencies of 31.4% and 29.3%, respectively. Hepatitis individuals were six times more likely to be HLA-A∗30:01 carriers than uninfected controls (OR = 6.30, p = 0.020 (HBV); OR = 6.21, p = 0.010 (HCV), respectively). Similarly, carriers of HLA-C∗17:01 were over-represented in the HBV-infected compared to the uninfected control group (21.9% vs. 6.4%, respectively) suggesting that this allele could play a role in the susceptibility to HBV infection.These findings demonstrate that carriers of HLA-A∗30:01 were over-represented in the hepatitis group compared to uninfected controls while HLA-C∗17:01 was completely absent in the HCV + group.

Metabolome Changes during InVivo Red Cell Aging Reveal Disruption of Key Metabolic Pathways.

SummaryUnderstanding the mechanisms for cellular aging is a fundamental question in biology. Normal red blood cells (RBCs) survive for approximately 100 days, and their survival is likely limited by functional decline secondary to cumulative damage to cell constituents, which may be reflected in altered metabolic capabilities. To investigate metabolic changes during in vivo RBC aging, labeled cell populations were purified at intervals and assessed for abundance of metabolic intermediates using mass spectrometry. A total of 167 metabolites were profiled and quantified from cell populations of defined ages. Older RBCs maintained ATP and redox charge states at the cost of altered activity of enzymatic pathways. Time-dependent changes were identified in metabolites related to maintenance of the redox state and membrane structure. These findings illuminate the differential metabolic pathway usage associated with normal cellular aging and identify potential biomarkers to determine average RBC age and rates of RBC turnover from a single blood sample.

Feasibility and cost of FH cascade screening in Belgium (BEL-CASCADE) including a novel rapid rule-out strategy

Background Familial hypercholesterolaemia (FH) is underdiagnosed in most countries. We report our first experience from a national pilot project of cascade screening in relatives of FH patients. Methodology Participating specialists recruited consecutive index patients (IP) with Dutch Lipid Clinic Network (DLCN) score ≥6. After informed consent, the relatives were visited by the nurses to collect relevant clinical data and perform blood sampling for lipid profile measurement. FH diagnosis in the relatives was based on the DLCN and/or MEDPED FH (Make-Early-Diagnosis-to-Prevent-Early-Deaths-in-FH) criteria. Results In a period of 18 months, a total of 127 IP (90 with definite FH and 37 with probable FH) were enrolled in 15 centres. Out of the 270 relatives visited by the nurses, 105 were suspected of having FH: 31 with DCLN score >8, 33 with DLCN score 5-8 and 41 with MEDPED FH criteria. In a post-hoc analysis, another set of MEDPED FH criteria established in the Netherlands and adapted to Belgium allowed to detect FH in 51 additional relatives. Conclusion In a country with no national FH screening program, our pilot project demonstrated that implementing a simple phenotypical FH cascade screening strategy using the collaboration of motivated specialists and two nurses, allowed to diagnose FH in 127 index patients and an additional 105 of their relatives over the two-year period. Newly developed MEDPED FH cut-offs, easily applicable by a nurse with a single blood sample, might further improve the sensitivity of detecting FH within families.

Pre- and post-diagnostic blood profiles of perfluoroalkyl acids in type 2 diabetes mellitus cases and controls

BackgroundStudies exploring the associations between perfluoroalkyl acids (PFAAs) and type 2 diabetes mellitus (T2DM) are rather limited and have reported conflicting results. All studies to date, including prospective ones, have relied on a single blood sample to study this association. Similarly, studies investigating how T2DM status may influence the longitudinal changes in PFAA concentrations have not been previously performed. As PFAA concentrations in humans have changed considerably over the last two decades, and as individuals diagnosed with T2DM usually undergo lifestyle changes that could influence these concentrations, a single blood sample may not necessarily reflect the life-time exposure to PFAA concentrations. Hence, repeated measurements from the same individuals will extend our understanding of how PFAAs are associated with T2DM. The present study, therefore, aimed to explore associations between pre- and post-diagnostic PFAA blood profiles and T2DM and assess factors associated with longitudinal changes in PFAAs in T2DM cases and controls. MethodsQuestionnaire data and blood samples from women participating in the Norwegian Women and Cancer study were used to conduct a nested case-control study among 46 T2DM cases matched to 85 non-diabetic controls. PFAAs were measured in blood samples collected prior to (2001/02) and after (2005/6) T2DM diagnosis. We investigated the association between PFAAs and incident and prevalent T2DM using conditional logistic regression. We assessed the longitudinal changes in PFAA concentrations within and between matched cases and controls using t-tests and linear regression models. ResultsWe observed no significant associations between pre-diagnostic PFAA concentrations and T2DM incidence. Similar results were observed for the post-diagnostic PFAA concentrations and T2DM prevalence. Decrease over time in PFAA concentrations were observed for PFOA and ∑PFOS concentrations, whereas increase over time were observed for PFNA, PFDA and PFUnDA concentrations. Longitudinal trends in PFAA concentrations among T2DM cases were similar to the changes observed in controls. ConclusionsThe study did not find evidence of association between PFAAs and incident or prevalent T2DM. The longitudinal changes in PFAAs concentrations were not influenced by T2DM status.

Abstract 4278: A composite neoantigen score is more strongly associated with therapeutic response than tumor mutational burden in a cohort of late-stage anti-PD-1-treated melanoma patients

Abstract Checkpoint inhibitor therapy has demonstrated meaningful, if varied antitumor activity, with patient response influenced by a variety of biological factors, including complex interactions between the tumor and immune system. Thus, it is of increasing interest to identify composite biomarkers integrating multiple biological features to better predict immunotherapy response. In this study we use a comprehensive tumor immungenomics profiling platform to examine the effectiveness of our composite neoantigen score for stratifying patient response to checkpoint blockade therapy compared to tumor mutational burden and other biomarkers.Pre-treatment tumor/normal samples from 55 unresectable, stage III/IV melanoma patients who underwent anti-PD-1 therapy were characterized to assess factors influencing response. RECIST criteria were used to evaluate tumor response to therapy, with a median follow-up of 18 months. For each patient, a single paired FFPE tumor and normal blood sample was collected and profiled using Personalis' ImmunoID NeXT platform; an augmented exome/transcriptome platform and analysis pipeline, which produces comprehensive tumor mutation information, gene expression quantification, neoantigen characterization, HLA typing and LOH, TCR repertoire profiling and tumor microenvironment profiling. These data were then analyzed together with clinical outcome, and a composite neoantigen score computed for each patient along with other biomarkers such as tumor mutational burden (TMB).In this cohort, an elevated pretreatment composite neoantigen score combining neoantigen predictions adjusted based on resistance mechanisms that affect neoantigen presentation on the MHC complex was more strongly predictive of response to PD-1 blockade than TMB alone. This was true for both response and non-response via RECIST criteria and progression free survival. We also found that the composite neoantigen score was a stronger predictor of patient response when compared to neoantigen burden alone. Additionally, we observed increased response to anti-PD-1 therapy in patients with elevated pretreatment TCR clonality. Combining the composite neoantigen score and TCR clonality data revealed a significant association with non-response to therapy. Patients with high composite neoantigen score and TCR clonality that failed to achieve complete response revealed potential resistance mechanisms to anti-PD-1 therapy. Specifically, we identified patients with high expression of IDO1 or CTLA4, which may facilitate PD-1-independent immune escape. Additionally, we found patients with mutations within the antigen presentation machinery (APM), likely leading to loss of surface expression of the proteins, and in the case of B2M mutations, improper HLA class I folding and antigen presentation. These APM mutations likely result in reduced neoantigen presentation in these patients, facilitating tumor escape.In summary, our composite neoantigen score which integrates multiple components of MHC class I presentation into a single score, is more significantly associated with response to therapy than individual biomarkers such as tumor mutational burden. These findings highlight the promise of composite biomarkers for the optimization of anti-PD-1 therapy patient selection. Citation Format: Charles W. Abbott, Eric Levy, Rachel Marty Pyke, Rena McClory, Sekwon Jang, Richard Chen, Sean Boyle. A composite neoantigen score is more strongly associated with therapeutic response than tumor mutational burden in a cohort of late-stage anti-PD-1-treated melanoma patients [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4278.

Assessment of Alteration of Serum Adiponectin Levels in Migraineurs: An Interventional Case Control Study.

Background and Objectives:Migraine is one of the major headache disorders. Epidemiological studies have shown its high prevalence and negative impact on personal and socioeconomic aspects. It is currently ranked 19th by the “World Health Organization” amongst all diseases, leading to disability worldwide. Inflammatory mediators, which include adipokines, have been analyzed in migraine pathophysiology. Nevertheless, their role is not well recognized. This study is aimed to assess serum high molecular weight adiponectin (HMW-ADP) levels in migraineurs: during the ictal phase, prior to, and postabortive treatment.Methods:This was a hospital-based interventional case-controlled study, checking the peripheral blood samples from migraineurs during an acute attack and after one hour of treatment with naproxen sodium (10–15 mg/kg). Age, sex, and BMI matched controls without headache were taken, and a single blood sample was drawn in them. HMW-ADP levels were evaluated by immunoassays.Results:A total of 120 patients which included 60 migraine patients along with 60 controls without headache were involved in the study. HMW-ADP was higher in migraine patients (9.89 ± 5.04 mcg/mL) than in patients without headache history (4.63 ± 2.98 mcg/mL; P = < .001); along with this, serum HMW-ADP (6.4 ± 4.09 mcg/mL; P = <.001) was found to be significantly lower in responders 60 min after acute abortive treatment.Conclusion:HMW-ADP levels were raised in migraineurs. Additionally, among responders following abortive treatment a considerable reduction in the levels was noted. These results recommend that the HMW-ADP might be a possible “novel biomarker of acute remedy response in acute migraineurs”.

Assessment of peripheral blood neutrophil respiratory burst, phagocytosis and apoptosis in obese non-insulin dysregulated horses

Obesity is a highly prevalent condition in horses. Dysfunctional neutrophil activity has been reported in metabolically healthy obese humans, but minimal data exist regarding horses. The present study evaluated the effect of obesity on apoptosis, phagocytosis and oxidative burst activity of peripheral blood neutrophils from lean and obese non-insulin dysregulated horses. Seven lean (BCS, body condition score 4–6/9) and five obese (BCS 8–9) horses were enrolled in the study. All animals underwent two metabolic tests (OGT, oral glucose test; IRT, insulin response test) before their selection to ensure their metabolic status (non-insulin dysregulated). A single blood sample was obtained from each horse, and a discontinuous density gradient was carried out to isolate neutrophils. Phagocytosis, apoptosis and reactive oxygen species (ROS) production assays were performed for each animal. All statistical analyses were performed with unpaired two-tailed t-tests. Results indicate that neutrophils from obese non-insulin dysregulated horses have a significantly increased ROS production (P < .0001), with no changes observed on phagocytosis (P > .05) or apoptosis (P > .05) when compared to the control group. In conclusion, our results demonstrate that obesity per se, in absence of other endocrine disorders, alters neutrophil reactive oxygen species production. More research is needed to understand the role of obesity on the equine immune system of horses, and its role in the development of endocrine disorders.

Abstract PR11: Multimodal analysis of circulating tumor cell RNA, circulating cell-free DNA, and genomic DNA from a single blood sample collected into a PAXgene Blood ccfDNA Tube*

Abstract Background: High demand for liquid biopsy tests in cancer research increases the need for combined approaches that allow multimodal testing from a limited blood volume. Although crosslinking-based fixatives allow for efficient preservation of the circulating cell-free DNA (ccfDNA) profile in whole blood, analysis of mRNA from circulating tumor cells (CTCs) is hampered. Currently, there are no blood collection tubes available that allow RNA analysis from CTCs after prolonged (&amp;gt;48 hours) storage. In this research study, we present a newly developed combined workflow for blood stabilization and subsequent analysis of CTC RNA, ccfDNA, and gDNA after prolonged (≥72 hours) storage. Methods: To establish the multimodal workflow, blood was collected from healthy donors into PAXgene Blood ccfDNA Tubes*, aliquoted and manually spiked with 20 individual LNCaP95 cells/5 mL blood as a CTC model or 20 µL PBS as a control. Blood samples were spiked and stored at 2–8°C for 3, 24, 48, and 72 hours before processing. CTCs were enriched and detected using the AdnaTest ProstateCancerPanel AR-V7† (QIAGEN). The plasma and cellular fraction of the CTC-depleted blood was then used to extract ccfDNA and gDNA, respectively. As a control for ccfDNA and gDNA yield measurements, plasma and the cellular fraction were also generated from whole samples (not CTC-depleted) spiked with 20 LNCaP95 cells/5 mL. The resulting multimodal workflow was tested: 1) in comparison to Streck Cell-Free DNA BCT; 2) to assess the impact of room temperature. Results: Tumor cells spiked into PAXgene Blood ccfDNA Tubes could be enriched and detected at experimental time points 3, 24, 48, and 72 hours after spiking with 100% test sensitivity within 24 hours storage and 91% test sensitivity after 72 hours storage at 2–8°C. In unspiked samples, the PAXgene stabilization solution did not cause false positive signals at any experimental time point. Yields of ccfDNA and gDNA were not statistically significantly affected by CTC capture (p&amp;gt;0.05). RNA from spiked tumor cells was detected in blood samples collected and stored in Streck Cell-Free DNA BCTs for 3 hours, but not at later time points. Storage of PAXgene stabilization solution-treated samples at room temperature is feasible, with 100% test sensitivity for CTC detection within 24 hours storage (similar to storage at 2–8°C) and 80% after 72 hours storage. Conclusions: Blood collected into PAXgene Blood ccfDNA Tubes and stored for up to 72 hours at 2–8°C or at room temperature can be used for multimodal analysis of CTC RNA, ccfDNA, and gDNA from a single blood sample. *For research use only. Not for use in diagnostic procedures. †For molecular biology applications. Not intended for the diagnosis, prevention, or treatment of a disease. This abstract is also being presented as Poster B18. Citation Format: Eric Provencher, Maike Remmel, Siegfried Hauch, Michael Otte, Andrea Ullius, Daniel Groelz, Anna Babayan. Multimodal analysis of circulating tumor cell RNA, circulating cell-free DNA, and genomic DNA from a single blood sample collected into a PAXgene Blood ccfDNA Tube* [abstract]. In: Proceedings of the AACR Special Conference on Advances in Liquid Biopsies; Jan 13-16, 2020; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2020;26(11_Suppl):Abstract nr PR11.

Urinary Concentrations of Phthalate Metabolite Mixtures in Relation to Serum Biomarkers of Thyroid Function and Autoimmunity among Women from a Fertility Center.

Background:Although previous epidemiological studies have explored associations of phthalate metabolites with thyroid function, no studies to date have assessed associations of mixtures with thyroid function and autoimmunity among potentially susceptible subgroups such as subfertile women.Objective:We aimed to explore associations of mixtures of urinary phthalate metabolites with serum markers of thyroid function and autoimmunity.Methods:This cross-sectional study included 558 women attending a fertility center who provided one spot urine and one blood sample at enrollment (2005–2015). We quantified urinary concentrations of eight phthalate metabolites using mass spectrometry, and biomarkers of thyroid function [thyroid-stimulating hormone (TSH), free and total thyroxine (, ) and triiodothyronine (, ), and autoimmunity [thyroid peroxidase and thyroglobulin antibodies (TPOAb and TgAb, respectively)] in serum using electrochemiluminescence assays. We applied principal component analysis (PCA) and Bayesian kernel machine regression (BKMR) to identify the main patterns of urinary phthalate metabolites. We used linear mixed models to assess the association between PCA-derived factor scores in quintiles and serum thyroid function and autoimmunity, adjusting for age, body mass index (BMI), specific gravity (SG), and, for the PCA, other factor scores.Results:We observed two factors using PCA, one representing the di(2-ethylhexyl) (DEHP) and another non-DEHP metabolites. Compared to women in the lowest quintile of the DEHP factor scores, women in the highest quintile had significantly lower serum concentrations of , , , and [absolute difference: ; 95% confidence interval (CI): , ; ; absolute difference: ; 95% CI: , ; ; absolute difference: ; 95% CI: 0.54, ; ; and absolute difference: ; 95% CI: , ; , respectively]. Using BKMR, we observed that mono(2-ethyl-5-hydroxyhexyl) phthalate (MEHHP) was the primary contributor to these negative associations. DEHP and non-DEHP factor scores were not associated with serum TSH, TgAb, or TPOAb.Conclusions:Mixtures of urinary DEHP metabolites were inversely associated with serum biomarkers of thyroid function but not with autoimmunity, which were within normal ranges for healthy adult women. https://doi.org/10.1289/EHP6740

P0193MEASUREMENTS OF SERUM CREATININE AND EGFR IN PATIENTS WITH CHRONIC CARDIORENAL SYNDROME TYPE 2 ARE INSUFFICIENT TO ASSESS RENAL RESPONSES TO RESYNCHRONIZATION THERAPY DURING A TREE-MONTH FOLLOW-UP

Abstract Background and Aims Chronic cardiorenal sydrome type 2 (CRS-T2) has a complex pathophysiology. Treatment and evaluation of the clinical effects of CRS-T2 therapy is difficult in clinical practice. The study’s aim was to non-invasively assess renal function using biomarkers associated with nephron injury sites in patients with CRS-T2 following cardiac resynchronization therapy (CRT). Method The study included patients with CRS-T2 and heart failure in NYHA classes II-IV, in whom CRT devices had been implanted. Before cardiac resynchronization therapy and after 3 months, patients’ renal function was assessed using biomarkers measured in single urine and blood samples. Test results were correlated with cardiovascular fitness assessments. Results CRT was implanted in 56 (n = 17; 30.4% ♀) patients with a mean age of 66 ± 10 years with CRS-T2 in the course of coronary artery disease (n = 38) or dilated cardiomyopathy (n = 18) with eGFRCKD-EPI = 68.55 ± 20.34 ml / min / 1.73m2. The three-month follow-up was continued in 33 (61.11%) patients. Three months after implantation CRT, the responders group cardiovascular endurance improved by at least on NYHA class; showed a significant decrease in NT-pro BNP (n = 24; 72.72%; p = 0.004), a significant (p &amp;lt; 0.05) decrease in serum prostaglandin D2 synthase (sPGD2S) and albuminuria (uACR) concentrations (Figure 1 a), and increases in hematocrit (HCT), erythrocyte count (RBC) and hemoglobin (Hb). The urine samples of the non-responders group following CRT, showed significant (p &amp;lt;0.05) increases in lipocalin concentrations associated with neutrophil gelatinase (uNGAL), and a decrease in cystatin C (uCysC) concentrations (Figure 1 b). There were no significant changes in the concentration of serum creatinine (sCr), eGFRCKD-EPI, serum cystatin C or uromodulin in urine and serum; and urinary kidney injury molecule-1 (uKIM-1) was found in the urine of both responding and nonresponding groups (p&amp;gt; 0.05). Conclusion The sCR and eGFR CKD-EPI assessment should not be used to assess renal function in patients with type 2 chronic cardiac renal syndrome who have undergone CRT implantation. Renal function biomarkers that account for the pathophysiology of nephron injury and correlated with myocardial function: sPGD2S, uACR, uCysC, uNGAL. Also, increased HCT, RBC, and Hb are characteristic in responders.

Clinical and analytical validation of FoundationOne Liquid CDx, a novel 324-gene blood-based comprehensive genomic profiling assay.

e13685 Background: As the availability of precision therapies expand, a well-validated blood-based comprehensive genomic profiling (CGP) assay has the potential to provide considerable value as a complement to tissue-based testing to ensure that potentially life-extending therapies are administered to the patients most likely to benefit. Comprehensive clinical and analytical validity data for blood-based assays are crucial to enabling physicians to understand the true performance of available testing options. Methods: The FoundationOne Liquid CDx assay is a blood-based CGP assay that has been validated for a Premarket Approval (PMA) submission to the Food and Drug Administration (FDA). Validation studies included &gt; 9,000 tests and &gt; 30,000 unique variants across &gt; 300 genes and &gt; 50 cancer types, allowing for a comprehensive assessment of performance. Results: The results of these extensive studies demonstrate a 95% limit of detection (LoD) of 0.40% variant allele fraction (VAF) for select short variants (subs and indels), 0.37% VAF for select rearrangements, 21.7% tumor fraction (TF) for select copy number amplifications, and 30.4% TF for copy number losses. LoD for complex biomarkers such as microsatellite instability (MSI) and blood tumor mutational burden (bTMB) were also determined. The limit of blank (LoB) was shown to be the ideal value of zero. Reproducibility of variant calling was determined to be 99.59% with two-sided exact CI of (99.58%, 99.60%). In comparison with an orthogonal method, an overall PPA (95% CI) of 96.3% (94.8, 97.4%) and NPA (95% CI) of &gt; 99.9% (99.9%, 100.0%) was observed. Critically, clinical validity and concordance with tissue-based CGP results across multiple tumor types were also evaluated. Conclusions: The results of these studies demonstrate that FoundationOne Liquid CDx accurately and reproducibly detects the major types of genomic alterations (short variants, rearrangements, and copy number alterations), as well as complex biomarkers, such as MSI, bTMB, and tumor fraction. These data demonstrate that the assay can identify genomic variants that may inform therapeutic decisions for cancer patients with any solid tumor using a single blood sample. Additionally, clinical validation results establish FoundationOne Liquid CDx as an additional tool for physicians in the therapeutic management of cancer patients.

Deletion of fetal-specific maternal CD4+ T cells during human pregnancy

Abstract Maternal T cell responses to fetal alloantigens are thought to play a critical role in facilitating successful pregnancy, but mechanisms of maternal-fetal tolerance remain largely unclear. Studies in murine models suggest that deletion of fetus-specific CD4+ T cells may contribute to tolerance; however, the role of deletion in human pregnancy is unknown. Here we use bulk TCR sequencing to assess changes in the maternal CD4+ T cell repertoire over the course of human pregnancy. Peripheral blood was collected from four primigravid women during the first and third trimesters, and a single peripheral blood sample was taken from their respective partners. Maternal CD4+ T cells were isolated using FACS, and TCR sequencing was performed on genomic DNA using the ImmunoSeqTM assay. The maternal alloreactive repertoire was defined using mixed lymphocyte reactions (MLRs) in which irradiated paternal PBMCs were used to stimulate maternal blood. An average of 32,077 (SD=9,609) unique CD4+ T cell clones/subject were identified at each timepoint. Within the small cohort examined in this study, we observed a notable increase in repertoire clonality in two of four women (average change in clonality=0.0045 vs. 0.0003). The two subjects that had an increase in CD4 clonality had a significant reduction in the number of alloreactive CD4 clones detected in the third trimester relative to the first whereas this number was effectively unchanged in the other two subjects (first:third=1:0.1±0.02 vs. 1:1.0±0.1). These findings suggest that substantial deletion of CD4+ alloreactive clones occurs in some women over the course of gestation. Deletion of alloreactive T cell clones may thus represent one mechanism of maternal-fetal tolerance in human pregnancy.

Reliable quantification of 18F-GE-180 PET neuroinflammation studies using an individually scaled population-based input function or late tissue-to-blood ratio

PurposeTracer kinetic modeling of tissue time activity curves and the individual input function based on arterial blood sampling and metabolite correction is the gold standard for quantitative characterization of microglia activation by PET with the translocator protein (TSPO) ligand 18F-GE-180. This study tested simplified methods for quantification of 18F-GE-180 PET.MethodsDynamic 18F-GE-180 PET with arterial blood sampling and metabolite correction was performed in five healthy volunteers and 20 liver-transplanted patients. Population-based input function templates were generated by averaging individual input functions normalized to the total area under the input function using a leave-one-out approach. Individual population-based input functions were obtained by scaling the input function template with the individual parent activity concentration of 18F-GE-180 in arterial plasma in a blood sample drawn at 27.5 min or by the individual administered tracer activity, respectively. The total 18F-GE-180 distribution volume (VT) was estimated in 12 regions-of-interest (ROIs) by the invasive Logan plot using the measured or the population-based input functions. Late ROI-to-whole-blood and ROI-to-cerebellum ratio were also computed.ResultsCorrelation with the reference VT (with individually measured input function) was very high for VT with the population-based input function scaled with the blood sample and for the ROI-to-whole-blood ratio (Pearson correlation coefficient = 0.989 ± 0.006 and 0.970 ± 0.005). The correlation was only moderate for VT with the population-based input function scaled with tracer activity dose and for the ROI-to-cerebellum ratio (0.653 ± 0.074 and 0.384 ± 0.177). Reference VT, population-based VT with scaling by the blood sample, and ROI-to-whole-blood ratio were sensitive to the TSPO gene polymorphism. Population-based VT with scaling to the administered tracer activity and the ROI-to-cerebellum ratio failed to detect a polymorphism effect.ConclusionThese results support the use of a population-based input function scaled with a single blood sample or the ROI-to-whole-blood ratio at a late time point for simplified quantitative analysis of 18F-GE-180 PET.

PD35-08 CAN THE INFLAMMATORY POTENTIAL OF DIET RATHER THAN THE INDIVIDUAL DIETARY COMPONENTS DISCRIMINATES BETWEEN CALCIUM OXALATE RENAL STONE PATIENTS AND HEALTHY CONTROLS?

You have accessJournal of UrologyStone Disease: Epidemiology & Evaluation III (PD35)1 Apr 2020PD35-08 CAN THE INFLAMMATORY POTENTIAL OF DIET RATHER THAN THE INDIVIDUAL DIETARY COMPONENTS DISCRIMINATES BETWEEN CALCIUM OXALATE RENAL STONE PATIENTS AND HEALTHY CONTROLS? Montserrat Arzoz Fàbregas*, Josep Roca Antonio, Maria Doladé Botías, Allen Rodgers, Nitin Shivappa, and James R Hebert Montserrat Arzoz Fàbregas*Montserrat Arzoz Fàbregas* More articles by this author , Josep Roca AntonioJosep Roca Antonio More articles by this author , Maria Doladé BotíasMaria Doladé Botías More articles by this author , Allen RodgersAllen Rodgers More articles by this author , Nitin ShivappaNitin Shivappa More articles by this author , and James R HebertJames R Hebert More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000000906.08AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Many studies have investigated the relationship between individual food groups and the risk of urolithiasis focusing on how diet influences the urinary risk factors for CaOx supersaturation and crystallization. However, diet also can play an important role in regulating chronic inflammation but what is lacking is a quantitative measure for assessing the inflammatory potential of an individual’s diet and whether such a measure is correlated with stone risk. The Dietary Inflammatory Index (DII®) is an analytical tool which provides such a measure. The aim of this study is to test the hypothesis that the inflammatory potential of habitual diet is higher in renal stone formers than in normal controls. METHODS: A total of 97 stone formers (SFs) and 63 controls (Cs), matched on age (± 5 years) and sex completed a semi-quantitative food frequency questionnaire from which nutrient composition was computed. These data were used to calculate the Dietary Inflammatory Index (DII®),which evaluates the inflammatory potential of the diet according to the pro- and anti-inflammatory properties of its components. To control the effect of energy intake, energy-adjusted DII (E-DII®) scores were calculated. A single blood sample was obtained from each participant (n=160), and two consecutive overnight (8h) urine samples were collected from a subset of these (n=59 SFs and n=54 Cs). Samples were analysed for stone risk factors. Data were analysed using descriptive statistics and logistic and linear regression models. RESULTS: DII and E-DII values were significantly more positive (i.e., more pro-inflammatory) in cases (-0.7 and -0.3) than in controls (-2.6 and -1.7), p<0.0001 and p<0.0001 respectively. Significant negative correlations were seen between E-DII score and blood HDL cholesterol (p=0.03) and urine volume (p=0.003), While significant positive correlations were seen between E-DII and urine concentrations of calcium (p=0.02), phosphate (p<0.0001), magnesium (p<0.0001) and uric acid (p=0.004). CONCLUSIONS: Our results show, for the first time, that holistic evaluation of the inflammatory potential of the whole diet rather than that of individual dietary components, can differentiate between SFs and Cs, thereby demonstrating the importance of pro-inflammatory diets in stone pathogenesis. As such, our study provides a compelling argument for determining DII/ E-DII scores in lithogenic patients on presentation and after dietary intervention, as part of their routine work-up. Source of Funding: Non financial support © 2020 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 203Issue Supplement 4April 2020Page: e720-e720 Advertisement Copyright & Permissions© 2020 by American Urological Association Education and Research, Inc.MetricsAuthor Information Montserrat Arzoz Fàbregas* More articles by this author Josep Roca Antonio More articles by this author Maria Doladé Botías More articles by this author Allen Rodgers More articles by this author Nitin Shivappa More articles by this author James R Hebert More articles by this author Expand All Advertisement PDF downloadLoading ...