Osong Public Health and Research Perspectives

Original Articles

Instability of Plasma and Serum Progastrin-Releasing Peptide During Repeated Freezing and Thawing: Jae-Eun Lee, Jin-Hyun Lee, Maria Hong, Seul-Ki Park, Ji-In Yu, So-Youn Shin, Shine Young Kim; Osong Public Health Res Perspect. 2016;7(6):351-355. Published online December 31, 2016; DOI: https://doi.org/10.1016/j.phrp.2016.11.004

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Objectives
Progastrin-releasing peptide (proGRP) is a promising biomarker for small cell lung cancer. However, not much is known about how sample processing and storage conditions affect the stability of proGRP. Here, we examined the effects of repeated freeze–thaw cycles on the stability of proGRP in plasma and serum.
Methods
Concentrations of proGRP were measured in plasma and serum samples exposed to two, three, or four freeze–thaw cycles and these were compared with values of corresponding samples exposed to one cycle (baseline). We also performed the area under the receiver-operating-characteristic curve (AUC) analysis to determine whether the differences of proGRP concentrations between each paired plasma and serum sample (ΔproGRP) can be used for identifying the samples that have been exposed to multiple freeze–thaw cycles.
Results
Concentrations of proGRP gradually decreased in both plasma and serum samples with increasing numbers of freeze–thaw cycles. Reduction rates of proGRP concentrations were greater in serum than in plasma samples and serum proGRP levels declined with statistical significance (p < 0.001) up to 10.1% after four freeze–thaw cycles. The ΔproGRP measurement showed fair accuracy (AUC = 0.741) for identifying samples that had been through four freeze–thaw cycles. The sensitivity was 82.8% and specificity was 62.1% at an optimal cut-off point of > 4.9.
Conclusion
Our study shows that the stability of circulating proGRP is affected in both plasma and serum samples by repeated freezing and thawing. We also show that ΔproGRP could be used for identifying paired plasma and serum samples subjected to multiple freeze–thaw cycles.

Citations

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Effect of Repeated Freeze–Thaw Cycles on Influenza Virus Antibodies
Alessandro Torelli, Elena Gianchecchi, Martina Monti, Pietro Piu, Irene Barneschi, Carolina Bonifazi, Rosa Coluccio, Luisa Ganfini, Luciano Michele La Magra, Silvia Marconi, Ginevra Marzucchi, Ramona Pace, Laura Palladino, Bernardo Biagi, Emanuele Montomo
Vaccines.2021; 9(3): 267. CrossRef
The influence of different blood samples treatment methods on pro-gastrin-releasing peptide
Huiqin Jiang, Ling Luo, Kang Xiong, Chengwen He, Huaizhou Wang, Yanghua Qin
Medicine.2019; 98(26): e16130. CrossRef

Inorganic Phosphorus and Potassium Are Putative Indicators of Delayed Separation of Whole Blood: Jae-Eun Lee, Maria Hong, Seul-Ki Park, Ji-In Yu, Jin-Sook Wang, Haewon Shin, Jong-Wan Kim, Bok-Ghee Han, So-Youn Shin; Osong Public Health Res Perspect. 2016;7(2):90-95. Published online April 30, 2016; DOI: https://doi.org/10.1016/j.phrp.2015.11.003

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Abstract PDF

Objectives
The delayed separation of whole blood can influence the concentrations of circulating blood components, including metabolites and cytokines. The aim of this study was to determine whether clinical-biochemistry analytes can be used to assess the delayed separation of whole blood.
Methods
We investigated the plasma and serum concentrations of five clinical-biochemistry analytes and free hemoglobin when the centrifugation of whole blood stored at 4°C or room temperature was delayed for 4 hours, 6 hours, 24 hours, or 48 hours, and compared the values with those of matched samples that had been centrifuged within 2 hours after whole-blood collection.
Results
The inorganic phosphorus (IP) levels in the plasma and serum samples were elevated ≥ 1.5-fold when whole-blood centrifugation was delayed at room temperature for 48 hours. Furthermore, the IP levels in the plasma samples showed excellent assessment accuracy [area under the receiver-operating-characteristic curve (AUC) > 0.9] after a 48-hour delay in whole-blood separation, and high sensitivity (100%) and specificity (95%) at an optimal cutoff point. The IP levels in the serum samples also exhibited good assessment accuracy (AUC > 0.8), and high sensitivity (81%) and specificity (100%). The potassium (K⁺) levels were elevated 1.4-fold in the serum samples following a 48-hour delay in whole-blood separation. The K⁺ levels showed excellent assessment accuracy (AUC > 0.9) following a 48-hour delay in whole-blood separation, and high sensitivity (95%) and specificity (91%) at an optimal cutoff point.
Conclusion
Our study showed that the IP and K⁺ levels in the plasma or serum samples could be considered as putative indicators to determine whether whole-blood separation had been delayed for extended periods.

Citations

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Effect of delayed centrifugation of whole blood on serum samples stability
Massimo Daves, Vincenzo Roccaforte, Michele Giacomi, Monica Riva, Maria Leitner, Stefan Platzgummer, Gertraud Goetsch, Giuseppe Lippi
La Rivista Italiana della Medicina di Laboratorio .2017; 13(1): 41. CrossRef
Instability of Plasma and Serum Progastrin-Releasing Peptide During Repeated Freezing and Thawing
Jae-Eun Lee, Jin-Hyun Lee, Maria Hong, Seul-Ki Park, Ji-In Yu, So-Youn Shin, Shine Young Kim
Osong Public Health and Research Perspectives.2016; 7(6): 351. CrossRef

Effect of Repeated Freezing and Thawing on Biomarker Stability in Plasma and Serum Samples: Jae-Eun Lee, Shine Young Kim, So-Youn Shin; Osong Public Health Res Perspect. 2015;6(6):357-362. Published online December 31, 2015; DOI: https://doi.org/10.1016/j.phrp.2015.11.005

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Abstract PDF

Objectives
The stability of circulating proteins can be affected by repeated freezing and thawing. The aim of our study was to identify the effect of repeated freezing and thawing on the plasma and serum concentrations of eight proteins [interferon-γ, interleukin (IL)-8, IL-15, IL-17A, matrix metalloproteinase (MMP)-7, tumor necrosis factor-α, vascular endothelial growth factor (VEGF), and VEGF receptor 2 (VEGF-R2)].
Methods
We assessed the concentration changes of these proteins in 30 plasma and serum samples subjected to three, four, or five freeze–thaw cycles, and compared these with the concentration changes in the samples that were subjected to two freeze–thaw cycles before analysis.
Results
Repeated freezing and thawing by up to five cycles did not modify the plasma and serum concentrations of interferon-γ, IL-8, and VEGF-R2, while levels of MMP-7, tumor necrosis factor-α, and VEGF were significantly changed in both plasma and serum samples. Moreover, MMP-7 and VEGF concentrations tended to increase with freeze–thaw cycles. They were more elevated in plasma samples (up to about 15%) than in serum samples (up to about 7%), suggesting that serum is the preferred sample type for the analysis of circulating proteins.
Conclusion
This is the first report on the effect of repeated freezing and thawing on plasma concentrations of MMP-7 and VEGF-R2. Our findings propose that researchers should consider the number of freeze–thaw cycles to select plasma or serum samples, depending on the type of analyte.

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Instability of Plasma and Serum Progastrin-Releasing Peptide During Repeated Freezing and Thawing
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Osong Public Health and Research Perspectives.2016; 7(6): 351. CrossRef
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Aging-related Changes in Mouse Serum Glycerophospholipid Profiles: Seungwoo Kim, Hyo-Soon Cheon, Jae-Chun Song, Sang-Moon Yun, Sang Ick Park, Jae-Pil Jeon; Osong Public Health Res Perspect. 2014;5(6):345-350. Published online December 31, 2014; DOI: https://doi.org/10.1016/j.phrp.2014.10.002

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Abstract PDF

Objectives
Metabolic dysfunction is a common hallmark of the aging process and aging-related pathogenesis. Blood metabolites have been used as biomarkers for many diseases, including cancers, complex chronic diseases, and neurodegenerative diseases.
Methods
In order to identify aging-related biomarkers from blood metabolites, we investigated the specific metabolite profiles of mouse sera from 4-month-old and 21-month-old mice by using a combined flow injection analysis–tandem mass spectrometry and liquid chromatography–tandem mass spectrometry.
Results
Among the 156 metabolites detected, serum levels of nine individual metabolites were found to vary with aging. Specifically, lysophosphatidylcholine (LPC) acyl (a) C24:0 levels in aged mice were decreased compared to that in young mice, whereas phosphatidylcholine (PC) acyl-alkyl (ae) C38:4, PC ae C40:4, and PC ae C42:1 levels were increased. Three classes of metabolites (amino acids, LPCs, and PCs) differed in intraclass correlation patterns of the individual metabolites between sera from young and aged mice. Additionally, the ratio of LPC a C24:0 to PC ae C38:4 was decreased in the aged mice, whereas the ratio of PC ae C40:4 to LPC a C24:0 was increased, supporting the aging-related metabolic changes of glycerophospholipids.
Conclusion
The ratios of the individual metabolites PC and LPC could serve as potential biomarkers for aging and aging-related diseases.

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Article

Serum MicroRNA Expression Profiling in Mice Infected with Rabies Virus: Myung Guk Han, Jun-Sun Park, Cho Soon Lee, Young Eui Jeong, Jung Sang Ryou, Jung Eun Cho, Young Ran Ju, Kyoung-Ki Lee; Osong Public Health Res Perspect. 2011;2(3):186-191. Published online December 31, 2011; DOI: https://doi.org/10.1016/j.phrp.2011.11.043

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Abstract PDF

Objectives
Serum or plasma microRNAs (miRNAs) are potential biomarkers for the diagnosis for cancer and prenatal diseases. This study was conducted to investigate whether rabies virus causes a change in serum miRNA expression.
Methods
ICR mice were intramuscularly inoculated with rabies virus and were sacrificed weekly to collect serum and brain tissue for 4 weeks postinoculation. Mice were assigned to four groups based on the results of indirect immunofluorescent assays, enzyme-linked immunosorbent assay, and nested reverse transcription-polymerase chain reaction and the expression profiles of serum miRNAs were compared using a commercial mouse miRNA expression profiling assay.
Results
The expression levels of miRNAs changed significantly with the different stages of the disease. The expression level of 94 serum miRNAs in infected mice changed at least twofold. Seven microRNAs of them were significantly upregulated or downregulated in all infected mice regardless of disease status. The number of miRNAs with an expression level change decreased with the progression of the disease. In a hierarchical cluster analysis, infected mice clustered into a group separate from uninfected control mice.
Conclusions
Based on the relationship of miRNAs to gene expression regulation, miRNAs may be candidates for the study of viral pathogenesis and could have potential as biomarkers.

Citations

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