Time Of Blood Collection Research Articles

Reduced plasma levels of RGM-A predict stroke-associated pneumonia in patients with acute ischemic stroke: A prospective clinical study.

BackgroundStroke-induced immunodepression syndrome is considered the major etiology of stroke-associated pneumonia (SAP). Repulsive guidance molecule A (RGM-A) is an immunomodulatory protein that is closely related to inflammation and immune responses. To explore the relationship between RGM-A and SAP and facilitate the early identification of patients at high risk of developing SAP, we investigated the predictive value of RGM-A in SAP.MethodsWe enrolled 178 patients with acute ischemic stroke (AIS) and finally analyzed 150 patients, among whom 69 had SAP and 81 had non-SAP. During the same period, 40 patients with community-acquired pneumonia and 40 healthy participants were included as controls. SAP was defined according to the modified US Centers for Disease Control and Prevention criteria. Blood samples were collected at 24 h, 48 h, 3 days, 4 to 7 days, and 8 to 14 days after stroke onset. An enzyme-linked immunosorbent assay was used to detect the plasma levels of RGM-A and interleukin-6.ResultsThe plasma RGM-A levels were significantly decreased in both patients with community-acquired pneumonia and those with AIS, and the decline was most pronounced in patients with SAP (P < 0.001). RGM-A started to decline within 24 h after stroke in the SAP group, and the lowest levels were detected on day 3 and days 4 to 7 (P < 0.001). The RGM-A levels in the SAP group were lower than those in the non-SAP group at all blood collection time points (P < 0.05). In the logistic regression analyses, RGM-A was a protective factor for SAP after adjusting for confounders (adjusted odds ratio = 0.22, 95% confidence interval = 0.091–0.538, P = 0.001). Receiver operating characteristic curve analysis showed that the area under the curve for RGM-A was 0.766 (0.091–0.538; P = 0.001), the cutoff value was 4.881 ng/mL, and the sensitivity and specificity were 80.00 and 76.36%, respectively.ConclusionsWe demonstrated that reduced plasma levels of RGM-A might help in the early identification of high-risk patients with SAP and predict the occurrence of SAP in patients with AIS. RGM-A might provide new clues to a potential alternative therapy for SAP.

Prediagnostic plasma polyphenol concentrations and colon cancer risk: The JPHC nested case–control study

Epidemiological studies that assessed the associations between dietary polyphenol intakes and colon cancer risk have reported largely null results, possibly due to measurement error associated with dietary assessment. We adopted an objective approach by measuring prediagnostic plasma concentrations of 35 polyphenols and assessing associations with colon cancer risk. We conducted a nested-case control study within the Japan Public Health Center-based prospective study (JPHC Study) utilizing plasma samples collected at the time of a five-year follow-up survey between 1995 and 1999. We identified colon cancer cases who developed cancer during the follow-up from the time of blood collection. Controls were matched by age, sex, area code, population size of the area, season of blood collection, year of blood collection, and duration of fasting time before the blood collection. Prediagnostic concentrations of 35 polyphenols from 375 incident colon cancer cases (followed until 2012) and 710 matched controls were measured by tandem mass spectrometry coupled with ultra-high-pressure liquid chromatography. We used multivariable conditional logistic regression models adjusted for established colon cancer risk factors to estimate odds ratios (ORs) and 95% confidence intervals (CIs). In sexes combined log2-transformed multivariable models, circulating levels of 3,4-dihydroxyphenylpropionic acid (P=0.02), ferulic acid (P=0.02), and caffeic acid (P=0.03) were inversely, and 3-hydroxybenzoic acid (P=0.03) was positively, associated with colon cancer risk. For men only, circulating levels of 3,4-dihydroxyphenylpropionic acid was inversely, and 3,5-dihydroxyphenylpropionic acid, gallic acid, (+)-epigallocatechin, 3-hydroxybenzoic acid, and epicatechin were positively, associated with colon cancer risk. In women, plasma caffeic acid and ferulic acid concentration were inversely associated with colon cancer risk. However, all these associations were nonsignificant after adjustment for multiple comparisons. The remaining polyphenols were not associated with colon cancer risk. Coffee-derived 3,4-dihydroxyphenylpropionic acid, ferulic acid, and caffeic acid concentrations were inversely associated with colon cancer risk although the association were nonsignificant after adjustment for multiple comparisons. These results support a possible role of coffee polyphenols in preventing colorectal cancer.

IGF-I Variability Over Repeated Measures in Patients With Acromegaly Under Long-Acting Somatostatin Receptor Ligands.

In patients with acromegaly on long-term treatment with long-acting somatostatin receptor ligands (SRLs), the time of blood collection for IGF-I measurement after injection is not well defined. We aimed to assess serum IGF-I dynamics and variability in SRL-treated patients compared with surgically cured patients and healthy controls. Thirty patients under SRLs considered controlled based on a normal previous IGF-I level, 10 patients cured by pituitary surgery, and 7 healthy subjects underwent 4 weekly IGF-I determinations. In SRL-treated patients, the IGF-I SDS (mean ± SD) was higher just before injection (0.34 ± 0.66) than at Day 7 (-0.33 ± 0.61; P = 0.0041) and Day 14 (-0.23 ± 0.60; P = 0.047) after injection, but it did not significantly vary in cured patients and healthy controls. The IGF-I CV was higher in SRL-treated patients than in cured patients or healthy controls (14.4 ± 7.6% vs 7.9 ± 4.4% and 8.3 ± 3.2%, respectively; P < 0.05 for both). Among SRL-treated patients, IGF-I CV was higher in "nonoptimally controlled patients"-i.e., patients with at least one elevated IGF-I value out of 4 (n = 9) compared with "optimally controlled" patients for whom all 4 IGF-I SDS values were < 2.0 (21.3 ± 9.3 vs 11.6 ± 6.0%; P = 0.0019). The latter did not differ from surgically cured patients and healthy controls. The measurement at the farthest distance from the SRL injection was the most predictive of patients with nonoptimally controlled disease. In patients treated with long-acting SRLs, IGF-I sampling at the farthest distance from SRL injection is the most informative and best predictor of optimal disease control.

Abstract LB547: Ultra-deep targeted sequencing of circulating tumor DNA with colorectal cancer patients for prediction of treatment response and post-surgical follow-up

Abstract Purpose: Colorectal cancer (CRC) patients require effective biomarkers to evaluate treatment efficacy and monitor minimal residual disease. However, current monitoring tools such as tumor markers and imaging methods have limited prognostic performance. Circulating-tumor DNA (ctDNA) could be a useful biomarker for monitoring patients with CRC. In this study, we evaluated the targeted next-generation sequencing (NGS) panel using ctDNA in monitoring clinical decision-making in CRC. Experimental Design: We prospectively enrolled 100 CRC patients and collected 197_ plasma samples. Specimens were collected post-surgery from 74 patients, 42 of whom were also monitored during the immediate post-surgery and adjuvant therapy follow-up. Twenty-six patients’ bloods were received after adjuvant therapy without surgery. A total of patients performed ultra-deep sequencing of CRC-related 14 genes respectively and matched with tissue samples at the time of blood collection as possible. NGS results were assessed correlation with clinicopathologic characteristics and clinical outcome. Results: Levels of ctDNA measurements were correlated with tumor stage. Stage IV patients had significantly higher ctDNA concentration than Stage I patients. ctDNA testing detected 112 alterations in 14 genes were identified in 69% of the patients (n=100). The results showed that APC (29%), TP53 (25%), KRAS (19%), and EGFR (5%) were the most frequently mutated genes. RAS family gene mutations were detected in 37 patients, yielding a concordance of 89% with tissue samples. Also, ctDNA detection was found to be a greater predictive performance when compared with CEA. Dynamics of ctDNA mutation levels reflected five follow-up patients’ clonal evolution and clinical outcomes. These data showed ctDNA is a sensitive monitoring biomarker for any clinical condition of the patients. Conclusions: The present study suggests that the use of ctDNA in CRC patients can be a valuable non-invasive monitoring tool for monitoring disease states during or after treatment. Serial ctDNA testing can contribute to improving patient management through monitoring disease progression and clonal evolution. Citation Format: Hyeonah Lee, Doo A Kim, Jieun Seo, Saeam Shin, Han Sang Kim, Joong Bae Ahn, Seung-Tae Lee, Jong Rak Choi. Ultra-deep targeted sequencing of circulating tumor DNA with colorectal cancer patients for prediction of treatment response and post-surgical follow-up [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr LB547.

Abstract 3401: Whole genome cell-free tumor DNA mutational signatures for noninvasive monitoring of pediatric brain cancers

Abstract Introduction: Liquid biopsy offers a noninvasive approach to monitor cancer burden during therapy and surveillance period. However, in pediatric brain cancers, liquid biopsy methods from the blood have been unsuccessful due to a low tumor burden and low number of mutations in coding regions. We hypothesized that a whole genome sequencing (WGS)-derived patient specific mutational signature from a matched tumor-normal WGS can provide a sensitive and specific approach to detect mutations in circulating cell free tumor DNA (ctDNA) and provide blood-based monitoring in pediatric patients with brain tumor. Methods: All tumors were analyzed and molecularly subclassified using whole genome DNA methylation profiling and machine learning classifier. Tumor DNA was extracted from pathology tissue and normal germline DNA from the white blood cells, while ctDNA was extracted from 1-2 mL of post-surgery or follow-up plasma samples, WGS was applied to sequence DNA from matched tumor-normal and plasma samples. WGS coverage was 40x for matched tumor-normal DNA and 20x for ctDNA. Using the C2i assay, we derived a personalized mutational pattern for each tumor and used an AI-based error suppression model for quantification and ultra-sensitive detection of ctDNA in plasma samples. A patient-specific personalized genome-wide compendium of somatic mutations was established and ctDNA tested at 1 to 3 available time points during the therapy or surveillance period. An AI-based error suppression model was implemented to filter out the noise in the cell free DNA (cfDNA) while the personalized mutational signature was used to detect the ctDNA in the cfDNA and to amplify the somatic signal contained in it. The ctDNA Tumor Fraction (TF) was compared to the clinical status and MR-based imaging. Results: We profiled 7 pediatric brain tumors, including 2 medulloblastomas (one Group 3, one Group 4), 3 pediatric glioblastomas IDH wild-type, 1 ependymoma PFA subtype and one low grade ganglioglioma. Tumor specific signatures were identified and detected in the plasma of 5 patients with clinical disease with a TF range 0.02-0.0005 but not in 2 patients with no tumor at the time of blood collection. In two children with a medulloblastoma and glioblastoma, the decrease of tumor fraction in ctDNA over 2 (TF: 0.002 to 0.0009) and 3 time points (TF: 0.0005 to undetectable), respectively, correlated with response to therapy based on imaging. Conclusions: Patient-specific WGS tumor signature in ctDNA from blood can be used for sensitive monitoring of children with brain tumors. Citation Format: Ivy Tran, Kristyn Galbraith, Guisheng Zhao, Robyn Borsuk, Joyce Varkey, Sharon Gardner, Jeffrey Allen, David Harter, Jeffrey Wisoff, Eveline T. Hidalgo, Sunil Deochand, Dillon Maloney, Danielle Afterman, Tomer Lauterman, Noah Friedman, Imane Bourzgui, Nidhi Ramaraj, Zohar Donenhirsh, Ronel Veksler, Jonathan Rosenfeld, Ravi Kandasamy, Iman Tavassoly, Boris Oklander, G. Praveen Raju, Theodore Nicolaides, Asaf Zviran, Matija Snuderl. Whole genome cell-free tumor DNA mutational signatures for noninvasive monitoring of pediatric brain cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3401.

Abstract 5896: Reproductive, hormonal and lifestyle correlates of circulating osteoprotegerin levels in women with a BRCA1 mutation

Abstract Purpose: BRCA1 mutation carriers face a very high lifetime risk of developing breast cancer. Dysregulation of the receptor activator of nuclear factor κB (RANK) pathway has been implicated in the pathogenesis of BRCA1-associated breast cancer. In addition, lower levels of osteoprotegerin (OPG), the decoy receptor for RANK-ligand, have been reported among women with a BRCA1 mutation and may be associated with an increased risk of breast cancer. Thus, inhibition of RANK signaling represents a potential target for prevention, and furthermore, OPG levels may be a biomarker of subsequent cancer risk. Whether non-genetic exposures also influence circulating levels of OPG in these high-risk women is not known. Therefore, the goal of this study was to evaluate reproductive, hormonal, and lifestyle correlates of OPG levels in BRCA1 mutation carriers. Methods: Eligible women included BRCA1 mutation carriers enrolled in a longitudinal study, aged 18 years or older, without a history of cancer, and with a serum sample available. All women completed a baseline questionnaire at the time of enrolment and a follow-up questionnaire every two years thereafter to collect detailed information on various exposures (i.e., reproductive, hormonal, and lifestyle) and outcomes. Serum OPG levels (pg/ml) were measured using an enzyme-linked immunosorbent assay (ELISA). Generalized linear models were used to evaluate the associations of various reproductive, hormonal, and lifestyle exposures at the time of blood collection with serum OPG and to estimate adjusted means. Results: A total of 828 women were included in the current analysis. Older age was associated with significantly higher OPG levels (&amp;lt;50 vs. &amp;gt;60 years, 79.04 vs. 82.01 pg/ml; Ptrend &amp;lt; 0.0001). Current vs. never smoking was also associated with significantly higher OPG levels (87.69 vs. 73.48 pg/ml; Pcat &amp;lt; 0.0001). There were no significant associations between other exposures and levels of OPG (P ≥ 0.17). Findings were similar in the analyses stratified by menopausal status. Conclusion: The results from this study suggest that non-genetic factors likely have a minimal impact upon serum OPG among women with a BRCA1 mutation. Further studies are needed to explore other potential correlates of OPG (i.e., diet, supplement use) and to elucidate whether integration of circulating OPG levels may improve existing risk prediction models. Citation Format: Sarah S. Park, Tasnim Zaman, Shana J. Kim, Jennifer D. Brooks, Andy K. Wong, Jan Lubiński, Steven A. Narod, Leonardo Salmena, Joanne Kotsopoulos. Reproductive, hormonal and lifestyle correlates of circulating osteoprotegerin levels in women with a BRCA1 mutation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5896.

Proteomics Studies in Gestational Diabetes Mellitus: A Systematic Review and Meta-Analysis.

Gestational Diabetes Mellitus (GDM) is the most common metabolic complication during pregnancy and is associated with serious maternal and fetal complications such as pre-eclampsia and stillbirth. Further, women with GDM have approximately 10 times higher risk of diabetes later in life. Children born to mothers with GDM also face a higher risk of childhood obesity and diabetes later in life. Early prediction/diagnosis of GDM leads to early interventions such as diet and lifestyle, which could mitigate the maternal and fetal complications associated with GDM. However, no biomarkers identified to date have been proven to be effective in the prediction/diagnosis of GDM. Proteomic approaches based on mass spectrometry have been applied in various fields of biomedical research to identify novel biomarkers. Although a number of proteomic studies in GDM now exist, a lack of a comprehensive and up-to-date meta-analysis makes it difficult for researchers to interpret the data in the existing literature. Thus, we undertook a systematic review and meta-analysis on proteomic studies and GDM. We searched MEDLINE, EMBASE, Web of Science and Scopus from inception to January 2022. We searched Medline, Embase, CINHAL and the Cochrane Library, which were searched from inception to February 2021. We included cohort, case-control and observational studies reporting original data investigating the development of GDM compared to a control group. Two independent reviewers selected eligible studies for meta-analysis. Data collection and analyses were performed by two independent reviewers. The PROSPERO registration number is CRD42020185951. Of 120 articles retrieved, 24 studies met the eligibility criteria, comparing a total of 1779 pregnant women (904 GDM and 875 controls). A total of 262 GDM candidate biomarkers (CBs) were identified, with 49 CBs reported in at least two studies. We found 22 highly replicable CBs that were significantly different (nine CBs were upregulated and 12 CBs downregulated) between women with GDM and controls across various proteomic platforms, sample types, blood fractions and time of blood collection and continents. We performed further analyses on blood (plasma/serum) CBs in early pregnancy (first and/or early second trimester) and included studies with more than nine samples (nine studies in total). We found that 11 CBs were significantly upregulated, and 13 CBs significantly downregulated in women with GDM compared to controls. Subsequent pathway analysis using Database for Annotation, Visualization and Integrated Discovery (DAVID) bioinformatics resources found that these CBs were most strongly linked to pathways related to complement and coagulation cascades. Our findings provide important insights and form a strong foundation for future validation studies to establish reliable biomarkers for GDM.

Circulating MicroRNAs as Biomarkers of Osteoporosis and Fragility Fractures.

Measurement of circulating microRNAs (miRNAs) as potential biomarkers of fragility fracture risk has recently become a subject of investigation. Measure by next-generation sequencing (NGS), global miRNA expression in serum samples of osteoporotic subjects vs individuals with normal bone mineral density (BMD). Samples were collected from patients with different bone phenotypes and/or fragility fractures who did not receive any antiresorptive and/or bone-forming drug at the time of blood collection. Samples and data were collected at 7 medical centers in Italy. NGS prescreening: 50 osteoporotic patients vs 30 individuals with normal BMD. Droplet digital polymerase chain reaction (ddPCR) validation: 213 patients with different bone phenotypes, including the NGS-analyzed cohort. NGS identified 5 miRNAs (miR-8085, miR-320a-3p, miR-23a-3p, miR-4497, miR-145-5p) differentially expressed in osteoporosis cases without fractures vs controls. ddPCR validation confirmed lower c-miR-23a-3p expression in osteoporotic patients, with or without fracture, than in osteopenic and normal subjects and increased c-miR-320a-3p expression in osteoporotic patients with fracture and lower expression in osteoporotic patients without fracture. ddPCR analysis showed a significantly increased expression of miR-21-5p in osteoporotic patients, with or without fracture, than in osteopenic and normal subjects, not evidenced by the NGS prescreening. Our study confirmed levels of c-miR-23a-3p and c-miR-21-5p as able to distinguish osteoporotic patients and subjects with normal BMD. Increased levels of c-miR-320a-3p specifically associated with fractures, independently by BMD, suggesting c-miR-320a-3p as a prognostic indicator of fracture risk in osteoporotic patients, to be confirmed in prospective studies on incident fractures.

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Comparison of Dechlorane Plus Concentrations in Sequential Blood Samples of Pregnant Women in Taizhou, China.

To develop an appropriate sampling strategy to assess the intrauterine exposure to dechlorane plus (DP), we investigated DP levels in sequential maternal blood samples collected in three trimesters of pregnancy, respectively, from women living in Taizhou. The median concentration of DPs (sum of syn-DP and anti-DP) in all samples was 30.5 pg g−1 wet-weight and 5.01 ng g−1 lipid-adjusted weight, respectively. The trimester-related DP concentrations were consistently strongly correlated (p < 0.01), indicating that a single measurement of DP levels could represent intrauterine exposure without sampling from the same female repeatedly; however, the wet-weight levels significantly increased across trimesters (p < 0.05), while the lipid-adjusted levels did not significantly vary. Notably, whether lipid-adjusted weight or wet-weight levels, the variation extent of DP across trimesters was found to be less than 41%, and those for other persistent organic pollutants (POPs) reported in the literature were also limited to 100%. The limitation in variation extents indicated that, regardless of the time of blood collection during pregnancy and how the levels were expressed, a single measurement could be extended to screen for exposure risk if necessary. Our study provides different strategies for sampling the maternal blood to serve the requirement for assessment of in utero exposure to DP.

Association Between Exposure to Per- and Polyfluoroalkyl Substances and Birth Outcomes: A Systematic Review and Meta-Analysis

BackgroundA large body of emerging evidence suggests that per- and polyfluoroalkyl substances (PFAS) affect birth outcomes in various pathways, but the evidence is inconsistent. Therefore, this study aimed to systematically review the epidemiological evidence on PFAS exposure and birth outcomes.MethodsThree electronic databases were searched for epidemiological studies through February 13, 2021. We used random-effects meta-analysis for eight birth outcome indicators to calculate summary effect estimates for various exposure types. The risk of bias and the overall quality and level of evidence for each exposure-outcome pair were assessed.ResultsThe initial search identified 58 potentially eligible studies, of which 46 were ultimately included. Many PFAS were found to have previously unrecognized statistically significant associations with birth outcomes. Specifically, birth weight (BW) was associated with PFAS, with effect sizes ranging from −181.209 g (95% confidence interval (CI) = −360.620 to −1.798) per 1 ng/ml increase in perfluoroheptanesulfonate (PFHpS) to −24.252 g (95% CI = −38.574 to −9.930) per 1 ln (ng/ml) increase in perfluorodecaoic acid (PFDA). Similar patterns were observed between other PFAS and birth outcomes: perfluorooctanoic acid (PFOA) and perfluorooctane sulfonate (PFOS) with birth length (BL) and ponderal index (PI), PFOS and perfluorododecanoic acid (PFDoDA) with head circumference (HC), PFHpS with gestational age (GA), and perfluorononanoic acid (PFNA) and PFHpS with preterm birth (PTB). Additionally, PFDA showed a statistically significant association with small for gestational age (SGA). The level of the combined evidence for each exposure-outcome pair was considered to be “moderate”.ConclusionThis study showed that PFAS exposure was significantly associated with increased risks of various adverse birth outcomes and that different birth outcome indicators had different degrees of sensitivity to PFAS. Further studies are needed to confirm our results by expanding the sample size, clarifying the effects of different types or doses of PFAS and the time of blood collection on birth outcomes, and fully considering the possible confounders.

Evaluation of antibody response after two doses of the Sinovac vaccine and the potential need for booster doses in cancer patients.

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is more severe in some specific patient groups, such as cancer patients with a mortality rate of 26.5%. The main way of protection is vaccination. In this study, we aimed to evaluate the antibody responses of our cancer patients who received two doses of the inactivated COVID-19 vaccine manufactured by Sinovac Life Sciences. Patients over the age of 18, who had not been suspected or polymerase chain reaction-confirmed COVID-19 positive, who received two doses of the vaccine, and at least 28 days passed after the second dose, and who received at least one dose of cancer treatment before the vaccination-were included in the study. Immunoglobulin class Gantibodies against the receptor binding region (S-RBD) of the spike protein S1 subunit of SARS-CoV-2 were studied. A total of 200 patients with a diagnosis of cancer were included in the study. The median time between the second dose of the Sinovac vaccine and the time of blood collection was 3.44 (3.20-3.84) months. SARS-CoV-2 antibody positivity was detected in 110 (55%) patients. The two subgroups with the highest antibody levels were gynecological cancers and breast cancers, with median 158.5 AU/ml(38.4-764.5) and 106.3 AU/ml(61.9-162.9) levels, respectively. Antibody positivity rate was 46.8% in patients who received chemotherapy at any time between the first dose of the vaccine and the date of blood collection; and it was 73.8% in the group that did not receive chemotherapy (p < 0.001). As a result, the expected antibody response was not obtained with two doses of the Sinovac vaccine. Therefore, if the Sinovac vaccine is to be preferred for these patients, the appropriate booster time for the third dose should be determined or other vaccines, such as messenger RNA vaccines, with reported higher antibody responses should be considered.

Pregnancy outcome prediction after embryo transfer based on serum human chorionic gonadotrophin concentrations.

The aim of the study was to evaluate the predictive value of the human chorionic gonadotropin (hCG) concentration on the 14th and 16th post-ovulation day after embryo transfer/cryoembryo transfer as well as the dynamics of its increase with respect to the outcome of pregnancy. In total, 130 embryo transfers and cryoembryo transfers in women aged 22 to 38 years who experienced a single embryo transfer or single cryoembryo transfer with confirmed pregnancy (hCG level over 15 IU/l on 14th post-ovulation day - D14) were selected. The input parameters (hCG D14, hCG D16, hCG D16-D14, hCG D16/D14 and positivity of at least 2.5-fold increase in hCG D16 compared to hCG D14) were evaluated by regression analysis in relation to the outcome parameters (bio-chemical pregnancy, clinical pregnancy, clinical pregnancy terminated by abortion up to 12 weeks of gestation, clinical pregnancy terminated by childbirth). Single concentrations of hCG D14 and D16, as well as the difference between these concentrations, were a statistically significant indicator of the prediction of bio-chemical pregnancy (P = 0.000215, P = 0.000227 and P = 0.000421). Contrary to expectations, the proportion of hCG D16 and D14 concentrations did not show statistical significance for either parameter, as well as the fulfilment of the condition of at least a 2.5fold increase in hCG D16 compared to D14. None of the studied input parameters was confirmed as a statistically significant marker for the prediction of miscarriage in the whole group of patients. However, in the group of confirmed clinical pregnancies, the serum concentration of hCG D16 (P = 0.0248) and the difference between concentrations D16 and D14 (P = 0.0185) were confirmed as a positive predictor of the progression of pregnancy until delivery. Single hCG concentrations are a good prognostic factor for predicting the outcome of pregnancy, but the determination of the cut-off limit is limited by inter-laboratory deviation as well as by timing of blood collection for hCG determination on the exact post-ovulatory day. The results of individual studies are therefore difficult to use in clinical practice. The dynamics of hCG concentrations appear to be a more reliable predictor of pregnancy outcome. In our cohort, we confirmed the statistical significance of the difference in hCG concentration between the 16th and 14th post-ovulation day not only for the prediction of bio-chemical pregnancy, but also as a predictor of the progression of clinical pregnancy into childbirth. To determine the optimal values of this difference, it is necessary to evaluate a larger group of patients. Conversely, the statistical significance of the proportion of hCG concentrations between the 16th and 14th post-ovulation day was not confirmed.

Characteristics of amino acid metabolism in preterm infants in Guangxi, China.

To study the characteristics of amino acid metabolism in preterm infants in Guangxi, China. A retrospective analysis was performed on the medical data of 30 757 neonates who underwent the screening for inherited metabolic diseases and had negative results in Guangxi Neonatal Disease Screening Center from 2018 to 2020. Among these neonates, there were 28 611 normal full-term infants (control group) and 2 146 preterm infants (preterm birth group). According to gestational age, the preterm infants were further divided into four groups: very preterm (n=209), moderately preterm (n=307), and late preterm group (n=1 630). According to birth weight, they were divided into three groups: very low birth weight group (n=161), low birth weight group (n=1 085), and normal birth weight group (n=900). According to blood collection time, they were divided into three groups: 3-7 days group (n=1 664), 8-14 days group (n=314) and 15-28 days group (n=168). Tandem mass spectrometry was performed to measure the levels of 11 amino acids in dried blood spots, which were then compared between groups. After adjustment for confounding factors, there were significant differences in the levels of 11 amino acids among different gestational age groups (P<0.05), and significant differences were observed in the levels of the 11 amino acids between the control group and the various preterm groups (except for citrulline and methionine in the late preterm group). There were significant differences in the levels of 11 amino acids among different birth weight groups (P<0.05). Except for ornithine, there were significant differences in the levels of other amino acids among the different blood collection time groups (P<0.05). Gestational age, birth weight and blood collection time all affect amino acid metabolism in preterm infants in Guangxi, China. This provides a basis for the laboratory to establish the reference standard and clinical interpretation of blood amino acid levels in preterm infants, and to improve the nutritional metabolism of preterm infants.

Prevalence of autoantibodies in patients with juvenile idiopathic arthritis: results from the German inception cohort ICON-JIA

BackgroundAn association of different autoimmune diseases is suspected. In juvenile idiopathic arthritis (JIA), only few and partially conflicting data on the co-existence of other autoimmune disorders are available. The prevalence of autoantibodies in patients with JIA in Germany is not known.MethodsSamples from 499 patients (median age at time of blood collection 11 years, median disease duration 4.4 years) in the prospective, multicenter inception cohort of children newly diagnosed with JIA (ICON-JIA) were analysed for the presence of anti-thyroid antibodies, celiac disease-specific antibodies (anti-tTG IgA, anti-tTG IgG), and connective tissue disease-associated antibodies (CTD-screen).ResultsA total of 76 (15.2%) patients had either clinically diagnosed autoimmune comorbidity or elevated autoantibodies. Of 21 patients with clinical autoimmune comorbidity, only 8 were also serologically positive at the time of testing, while 55 patients had autoantibodies without clinical diagnosis. Thus, 63 patients (12.6%) had at least one elevated autoantibody. Antibodies against thyroglobulin were found in 3% and against thyreoperoxidase in 4% of the samples. TSH receptor antibodies could not be detected in any of the 499 patients. Tissue transglutaminase antibodies were elevated in 0.4% of the patients. A positive screen for CTD-specific antinuclear antibodies was found in 7%, but only rarely specific antibodies (anti-dsDNA 1.4%, anti-SS-A and -SS-B 0.2% each, anti-CENP-B 0.4%) were confirmed.ConclusionsIn our study, a specific correlation between JIA and other autoimmune phenomena could not be confirmed. The lack of well-matched control groups makes interpretation challenging. Further data need to corroborate the suspected increased risk of developing other autoimmune phenomena in JIA patients.

Interferon signature in giant cell arteritis aortitis

ObjectivesMolecular mechanisms underlying large-vessel involvement in giant cell arteritis (LV-GCA) are largely unknown. Herein, we explore the critical involvement of pro-inflammatory signaling pathways in both aorta and T cells from patients with LV-GCA. MethodsWe analyzed transcriptome and interferon gene signature in inflamed aortas from LV-GCA patients and compared them to non-inflammatory control aorta. Differential transcriptomic analyses of circulating CD4+ and CD8+ T cells were also performed between patients with active GCA (not under any immunosuppressants or corticosteroid doses higher than 10 mg/day by the time of blood collection) and healthy donors. Interferon-alpha serum levels were measured using ultra-sensitive technique (HD-X Simoa Planar Technology) in GCA patients according to disease activity status. ResultsTranscriptomic analyses revealed 1042, 1479 and 2075 significantly dysregulated genes for aortas, CD4+ and CD8+ cells from LV-GCA patients, respectively, as compared to controls. A great enrichment for pathways linked to interferons (type I, II and III), JAK/STAT signaling, cytokines and chemokines was seen across aortas and circulating T cells. A type I interferon signature was identified as significantly upregulated in the aorta of patients with LV-GCA, notably regarding EPSTI1 and IFI44L genes. STAT3 was significantly upregulated in both aorta and T cells and appeared as central in related gene networks from LV-GCA patients. Interferon-alpha serum levels were higher in patients with active GCA when compared to those in remission (0.024 vs. 0.011 pg/mL; p = 0.028). ConclusionLV-GCA presents a clear type I interferon signature in aortas, which paves the way for tailored therapeutical targeting.

Estimation of Contrast Agent Concentration in DCE-MRI Using 2 Flip Angles.

The aim of this study was to investigate the feasibility of using 2 flip angles (FAs) with an ultrashort echo time during dynamic contrast-enhanced (DCE)-magnetic resonance imaging (MRI) for estimation of plasma gadolinium (Gd) concentration without using a precontrast longitudinal relaxation time T1 (T10) measurement. T1-weighted DCE-MRI experiments were carried out with C57BL/6J mice using the scan protocol with 2 FAs over 3 sequential segments during 1 scan. The data with 2 FAs were used to estimate T10 (T1T) during conversion of a time-intensity curve to the time-concentration curve. Three dosages of gadolinium-based contrast agent were used to achieve a wide range of variability in Gd concentrations when measured at 10 minutes postinjection: 0.05 mmol/kg (n = 6), 0.1 mmol/kg (n = 11), and 0.15 mmol/kg (n = 7). For comparison, the signal-to-concentration conversion was also conducted using the T10 measured from the precontrast scan (T1M) as well as a constant T10 (2.1 seconds) from the literature (T1C). The Gd concentrations ([Gd]) estimated using DCE-MRI data for the time of retro-orbital blood collection ([Gd]T1T, [Gd]T1M, and [Gd]T1C, respectively) were compared against the [Gd] of the blood samples measured by inductively coupled plasma mass spectrometry ([Gd]MS). In addition, contrast kinetic model analysis was conducted on mice with GL261 brain tumors (n = 5) using the 3 different methods for T10. T1T strongly correlated with T1M (r = 0.81). [Gd]T1M and [Gd]T1T were significantly different from [Gd]T1C. [Gd]T1M and [Gd]T1T were in good agreement with [Gd]MS with strong correlations (mean percentage error ± standard deviation) of r = 0.70 (16% ± 56%) and r = 0.85 (15% ± 44%), respectively. In contrast, [Gd]T1C had a weak correlation of r = 0.52 with larger errors of 33% ± 24%. The contrast kinetic model parameters of GL261 brain tumors using T1T were not significantly different from those using T1M. This study substantiates the feasibility of using the 2-FA approach during DCE-MRI scan to estimate [Gd] in the plasma without using an extra scan to perform precontrast T1 measurements.

Functional Effect of Lesser Yam Symbiotic Yoghurt on Hypercholesterolemia Wistar Rats

Synbiotic lesser yam yoghurt is fermented drink made from the mixture of cow’s milk and lesser yam extract which has functional benefits. Lesser yam yoghurt contains lactic acid bacteria, lactic acid, inulin, and soluble fiber which can improve the lipid profile of hypercholesterolemia sufferers. This study aims to determine the effect of the intake of lesser yam yoghurt on the lipid profile of hypercholesterolemia wistar rats. This study used nested randomized design with two factors, the first factor was a feeding group (0,1,2,3,4 ml/day, while the second factor was the time of blood collection (0, week 1 and week 2). The data obtained were analyzed by using ANAVA and further test DMRT (Duncan Multiple Range Test) with a level of 5%. The results showed that giving lesser yam yoghurt for 2 weeks had a significant effect on the lipid profile improvement. Rats from 4 ml of yoghurt treatment experienced a decrease in total serum cholesterol by 48.4%, triglyceride levels by 35.8% and LDL levels by 65,5% and an increase in serum HDL levels by 150.3%. So it can be concluded that the intake of synbiotik lesser yam yoghurt has functional benefits in improving the lipid profile of hypercholesterolemia rats.

Evaluation and follow-up of antibody formation after CoronaVac vaccine.

The aim of this study was to monitor the time-dependent change by evaluating the antibody levels at the 4th, 7th, 10th, 13th, and 16th weeks after the second dose of the CoronaVac vaccine. The study group (n=65) were between 21 and 60 years old and received two doses of the CoronaVac vaccine. Blood samples were collected after 4th, 7th, 10th, 13th, and 16th weeks of the second dose of the vaccine administration. There was a coronavirus disease 2019 recovered group (n=29) who were SARS-CoV-2 real-time PCR test result positive before the vaccination period, and no coronavirus disease 2019 history group (n=36). Age, BMI, gender, smoking, comorbidity, coronavirus disease 2019 contact history, and working in the coronavirus disease 2019 service history of the individuals were recorded. No statistically significant difference was found in the descriptive findings of the individuals according to coronavirus disease 2019 recovered group and no coronavirus disease 2019 history group. It was observed that antibody levels in the coronavirus disease 2019 recovered group were found to be higher for each period of serum collection compared to the no coronavirus disease 2019 history group, which were statistically significant. The distribution curves of the antibody levels according to the timing of blood collection in coronavirus disease 2019 recovered group, no coronavirus disease 2019 history group, and total subjects were extrapolated, and it was observed that the estimated time for the antibodies to reach the threshold value of the test was 214, 145, and 166 days after vaccination. It is important to make booster doses, as the CoronaVac vaccine will lose its effect after the fifth month due to the decrease in Ab levels. In addition, since the antibody levels decrease later in those who have a history of coronavirus disease 2019 infection and are vaccinated, individuals who have no previous history of coronavirus disease 2019 should be given priority for vaccination.

The influence of immunomodulators on the morpho biochemical status and development of calves in early postnatal ontogenesis

The influence of early immunological stimulation on the dynamics of the morphobiochemical composition of blood and the increase in live weight in calves of the black-and-white Holstein breed was evaluated. For the experiment, 3 groups of young animals were formed at the age of 7 days, 10 calves in each. The first group of animals was injected with 500 UNITS of bovine recombinant interferon-a2b, the second group used denatured emulsified placenta (PDE) of 5 ml, the third group of calves served as a control, where saline solution was used. The treatments were performed twice with an interval of 7 days. Blood collection was carried out before injections, on 7 and 14 days after the start of the experiment. After a single admin istration of interferon, an increase in urea level was observed by 44.7% (P&lt; 0.05), while the indicator was higher by 13.6...37.5% (P &lt; 0.01) in relation to the group of calves treated with PDE. The activity of transaminases in the experimental groups did not have significant dynamics, when in control calves AST increased by 28.9% (P≤0.05), and ALT by 2.3...2.7 times (P≤0.05...0.01). The activity of alkaline phosphatase in calves treated with PDE and saline solution was stable, in the first group the indicator decreased by 16.2...17.6% (P≤0.05...0.01), which is less by 21.8% (P≤0.05) in relation to the group of calves treated with PDE and by 45.5% (P≤0.05) in relation to the control. In the first group of calves, there was a decrease in total (by 46.7%, P &lt; 0.05) and free bilirubin (by 37.2%, P &lt; 0.01). The total bilirubin decreased by 47.5% (P &lt; 0.01) in the young animals injected with PDE, while the indicator increased slightly in the control group. Under the influence of interferon, BASC increased by 30.9% (P &lt; 0.05) after the first injection and by 43.2% (P &lt; 0.05) after the second injection. Double administration of PDE contributed to an increase in BASC by 42.0% (P≤0.05). LASC in the first group increased by 47.1% (P≤0.05), which is higher than the control animals by 19.0...38.9% (P≤0.05). In the third group, there was also an increase in WEASELS at the time of the last blood collection (by 40.0%, P&lt;0.05), while in calves that were injected with PDE, the increase in the indicator was not significant. The increase in live weight in the first three months of life during the use of interferon was greater by 7.3 kg or 12.7% (P &gt;&lt; 0.05) compared to the control group and by 3.2 kg or 5.1% compared to the group treated with PDE.