scholarly journals Integrating Whole Blood Transcriptomic Collection Procedures Into the Current Anti-Doping Testing System, Including Long-Term Storage and Re-Testing of Anti-Doping Samples

2021 ◽  
Vol 8 ◽  
Author(s):  
Giscard Lima ◽  
Alexander Kolliari-Turner ◽  
Fernanda Rossell Malinsky ◽  
Fergus M. Guppy ◽  
Renan Paulo Martin ◽  
...  

Introduction: Recombinant human erythropoietin (rHuEPO) administration studies involving transcriptomic approaches have demonstrated a gene expression signature that could aid blood doping detection. However, current anti-doping testing does not involve collecting whole blood into tubes with RNA preservative. This study investigated if whole blood in long-term storage and whole blood left over from standard hematological testing in short-term storage could be used for transcriptomic analysis despite lacking RNA preservation.Methods: Whole blood samples were collected from twelve and fourteen healthy nonathletic males, for long-term and short-term storage experiments. Long-term storage involved whole blood collected into Tempus™ tubes and K2EDTA tubes and subjected to long-term (i.e., ‒80°C) storage and RNA extracted. Short-term storage involved whole blood collected into K2EDTA tubes and stored at 4°C for 6‒48 h and then incubated at room temperature for 1 and 2 h prior to addition of RNA preservative. RNA quantity, purity, and integrity were analyzed in addition to RNA-Seq using the MGI DNBSEQ-G400 on RNA from both the short- and long-term storage studies. Genes presenting a fold change (FC) of >1.1 or < ‒1.1 with p ≤ 0.05 for each comparison were considered differentially expressed. Microarray analysis using the Affymetrix GeneChip® Human Transcriptome 2.0 Array was additionally conducted on RNA from the short-term study with a false discovery ratio (FDR) of ≤0.05 and an FC of >1.1 or < ‒1.1 applied to identify differentially expressed genes.Results: RNA quantity, purity, and integrity from whole blood subjected to short- and long-term storage were sufficient for gene expression analysis. Long-term storage: when comparing blood tubes with and without RNA preservation 4,058 transcripts (6% of coding and non-coding transcripts) were differentially expressed using microarray and 658 genes (3.4% of mapped genes) were differentially expressed using RNA-Seq. Short-term storage: mean RNA integrity and yield were not significantly different at any of the time points. RNA-Seq analysis revealed a very small number of differentially expressed genes (70 or 1.37% of mapped genes) when comparing samples stored between 6 and 48 h without RNA preservative. None of the genes previously identified in rHuEPO administration studies were differently expressed in either long- or short-term storage experiments.Conclusion: RNA quantity, purity, and integrity were not significantly compromised from short- or long-term storage in blood storage tubes lacking RNA stabilization, indicating that transcriptomic analysis could be conducted using anti-doping samples collected or biobanked without RNA preservation.

HortScience ◽  
2019 ◽  
Vol 54 (1) ◽  
pp. 143-148 ◽  
Author(s):  
Brianna L. Ewing ◽  
Gregory M. Peck ◽  
Sihui Ma ◽  
Andrew P. Neilson ◽  
Amanda C. Stewart

Hard cider production in the United States has increased dramatically during the past decade, but there is little information on how harvest and postharvest practices affect the chemistry of the resulting cider, including concentrations of organoleptically important flavanols. For 2 years we assessed fruit, juice, and cider from a total of five apple (Malus ×domestica Borkh.) cultivars in two experiments: sequential harvests and postharvest storage. Different cultivars were used in 2015 and 2016 with the exception of ‘Dabinett’, which was assessed in both years. There were no differences in polyphenol concentrations in cider made from fruit that was harvested on three separate occasions over a 4-week period in either 2015 or 2016. Fruit storage durations and temperatures had little influence on the chemistry when the experiment was conducted in 2015, but polyphenol concentration was greater after storage in the 2016 experiment. In 2016, total polyphenols in ‘Dabinett’ ciders were 51% greater after short-term storage at 10 °C and 67% greater after long-term storage at 1 °C than the control, which was not subjected to a storage treatment. In 2016, total polyphenols in ‘Binet Rouge’ ciders were 67% greater after short-term storage at 10 °C and 94% greater after long-term storage at 1 °C than the control. Although results varied among cultivars and harvest years, storing apples for longer periods of time and at warmer temperatures may be a strategy to increase polyphenol, particularly flavanol, concentrations in hard cider.


2022 ◽  
Vol 11 (1) ◽  
pp. 245
Author(s):  
Jan A. Graw ◽  
Victoria Bünger ◽  
Lorenz A. Materne ◽  
Alexander Krannich ◽  
Felix Balzer ◽  
...  

Packed red blood cells (PRBCs), stored for prolonged intervals, might contribute to adverse clinical outcomes in critically ill patients. In this study, short-term outcome after transfusion of PRBCs of two storage duration periods was analyzed in patients with Acute Respiratory Distress Syndrome (ARDS). Patients who received transfusions of PRBCs were identified from a cohort of 1044 ARDS patients. Patients were grouped according to the mean storage age of all transfused units. Patients transfused with PRBCs of a mean storage age ≤ 28 days were compared to patients transfused with PRBCs of a mean storage age > 28 days. The primary endpoint was 28-day mortality. Secondary endpoints included failure-free days composites. Two hundred and eighty-three patients were eligible for analysis. Patients in the short-term storage group had similar baseline characteristics and received a similar amount of PRBC units compared with patients in the long-term storage group (five units (IQR, 3–10) vs. four units (2–8), p = 0.14). The mean storage age in the short-term storage group was 20 (±5.4) days compared with 32 (±3.1) days in the long-term storage group (mean difference 12 days (95%-CI, 11–13)). There was no difference in 28-day mortality between the short-term storage group compared with the long-term storage group (hazard ratio, 1.36 (95%-CI, 0.84–2.21), p = 0.21). While there were no differences in ventilator-free, sedation-free, and vasopressor-free days composites, patients in the long-term storage group compared with patients in the short-term storage group had a 75% lower chance for successful weaning from renal replacement therapy (RRT) within 28 days after ARDS onset (subdistribution hazard ratio, 0.24 (95%-CI, 0.1–0.55), p < 0.001). Further analysis indicated that even a single PRBC unit stored for more than 28 days decreased the chance for successful weaning from RRT. Prolonged storage of PRBCs was not associated with a higher mortality in adults with ARDS. However, transfusion of long-term stored PRBCs was associated with prolonged dependence of RRT in critically ill patients with an ARDS.


1989 ◽  
Vol 61 (2) ◽  
pp. 373-385 ◽  
Author(s):  
S. R. Gooneratne ◽  
B. Laarveld ◽  
R. K. Chaplin ◽  
D. A. Christensen

1. The effectiveness of tetrathiomolybdate (TTM) in the removal of 67Cu from the long-term storage compartment in liver was studied. Lambs receiving 5 mg Cu/kg dry matter (DM) or 35 mg Cu/kg DM were primed intravenously (iv) with 67Cu and challenged 10 d later with 99Mo-labelled TTM given either iv or intraduodenally (id). The profiles of 67Cu and 99Mo and of Cu and Mo with time were measured in blood, bile, urine and faeces.2. The level of dietary Cu affected the amplitude of profiles of 67Cu and Cu in blood, bile and urine after administration of 99Mo-labelled TTM. TTM administration increased liver Cu removal and this was most marked in sheep given TTM iv. The liver Cu removal from the long-term storage Cu compartment was low and was not affected by the route of administration of TTM. Endogenous Cu excretion was higher in lambs given TTM id.3. Excretion of 67Cu in bile through the transhepatocellular pathway after TTM administration appeared absent, while the transbiliary and hepatolysosomal pathways were operative. The potential reasons for this change are discussed.4. TTM predominantly enhances the removal of Cu from the short-term storage compartment, but effects on the long-term storage compartment may still be of significance.


1971 ◽  
Vol 29 (3_suppl) ◽  
pp. 1179-1182 ◽  
Author(s):  
Beth J. Raymond

The present study was undertaken in order to examine output from short-term storage (STS) and long-term storage (LTS) in elderly Ss using a free-recall paradigm. Contrary to the widespread belief that aged Ss have difficulty recalling recent information, the data indicated no deterioration in recall from STS although recall from LTS was considerably less than is usually demonstrated in free-recall studies.


Author(s):  
Judith E. Heida ◽  
Lianne S.M. Boesten ◽  
Esmée M. Ettema ◽  
Anneke C. Muller Kobold ◽  
Casper F.M. Franssen ◽  
...  

AbstractBackground:Copeptin, part of the vasopressin precursor, is increasingly used as marker for vasopressin and is claimed to have better ex vivo stability. However, no study has directly compared the ex vivo stability of copeptin and vasopressin.Methods:Blood of ten healthy volunteers was collected in EDTA tubes. Next, we studied the effect of various pre-analytical conditions on measured vasopressin and copeptin levels: centrifugation speed, short-term storage temperature and differences between whole blood and plasma, long-term storage temperature and repeated freezing and thawing. The acceptable change limit (ACL), indicating the maximal percentage change that can be explained by assay variability, was used as cut-off to determine changes in vasopressin and copeptin.Results:The ACL was 25% for vasopressin and 19% for copeptin. Higher centrifugation speed resulted in lower vasopressin levels, whereas copeptin concentration was unaffected. In whole blood, vasopressin was stable up to 2 h at 25°C and 6 h at 4°C. In plasma, vasopressin was stable up to 6 h at 25°C and 24 h at 4°C. In contrast, copeptin was stable in whole blood and plasma for at least 24h at both temperatures. At –20°C, vasopressin was stable up to 1 month and copeptin for at least 4 months. Both vasopressin and copeptin were stable after 4 months when stored at –80°C and –150°C. Vasopressin concentration decreased after four freeze-thaw cycles, whereas copeptin concentration was unaffected.Conclusion:Vasopressin levels were considerably affected by pre-analytical conditions, while copeptin levels were stable. Therefore, a strict sample handling protocol for measurement of vasopressin is recommended.


Sign in / Sign up

Export Citation Format

Share Document