Effect of Different Anticoagulant Agents on Immune-Related Genes in Leukocytes Isolated from the Whole-Blood of Holstein Cows
Anticoagulants, such as ethylenediaminetetraacetic acid (EDTA), sodium citrate (Na-citrate), or heparin are normally used in hematological clinical tests to prevent coagulation. Although anticoagulants are fundamental for the correct application of clinical tests, they produce adverse effects in dif...
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MDPI AG
2023-02-01
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| Series: | Genes |
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| Online Access: | https://www.mdpi.com/2073-4425/14/2/406 |
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| author | Viviana Floridia Marta Sfulcini Enrico D’Alessandro Luca Cattaneo Matteo Mezzetti Luigi Liotta Erminio Trevisi Vincenzo Lopreiato Andrea Minuti |
| author_facet | Viviana Floridia Marta Sfulcini Enrico D’Alessandro Luca Cattaneo Matteo Mezzetti Luigi Liotta Erminio Trevisi Vincenzo Lopreiato Andrea Minuti |
| author_sort | Viviana Floridia |
| collection | DOAJ |
| description | Anticoagulants, such as ethylenediaminetetraacetic acid (EDTA), sodium citrate (Na-citrate), or heparin are normally used in hematological clinical tests to prevent coagulation. Although anticoagulants are fundamental for the correct application of clinical tests, they produce adverse effects in different fields, such as those involving specific molecular techniques; for instance, quantitative real time polymerase chain reactions (qPCR) and gene expression evaluation. For this reason, the aim of this study was to evaluate the expression of 14 genes in leukocytes that were isolated from the blood of Holstein cows, and which were collected in Li-heparin, K-EDTA, or Na-citrate tubes; then, they were analyzed using qPCR. Only the <i>SDHA</i> gene showed a significant dependence (<i>p</i> ≤ 0.05) on the anticoagulant that was used with the lowest expression; this was observed in Na-Citrate after being compared with Li-heparin and K-EDTA (<i>p</i> < 0.05). Although a variation in transcript abundance with the three anticoagulants was observed in almost all the investigated genes, the relative abundance levels were not statistically significant. In conclusion, the qPCR results were not influenced by the presence of the anticoagulant; thus, we had the opportunity to choose the test tube that was used in the experiment without interfering effects impacting the gene expression levels caused by the anticoagulant. |
| first_indexed | 2024-03-11T08:46:43Z |
| format | Article |
| id | doaj.art-3b4b5f0c84a648f0bde28fea19f5a777 |
| institution | Directory Open Access Journal |
| issn | 2073-4425 |
| language | English |
| last_indexed | 2024-03-11T08:46:43Z |
| publishDate | 2023-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Genes |
| spelling | doaj.art-3b4b5f0c84a648f0bde28fea19f5a7772023-11-16T20:42:32ZengMDPI AGGenes2073-44252023-02-0114240610.3390/genes14020406Effect of Different Anticoagulant Agents on Immune-Related Genes in Leukocytes Isolated from the Whole-Blood of Holstein CowsViviana Floridia0Marta Sfulcini1Enrico D’Alessandro2Luca Cattaneo3Matteo Mezzetti4Luigi Liotta5Erminio Trevisi6Vincenzo Lopreiato7Andrea Minuti8Department of Veterinary Sciences, Università di Messina, Viale Palatucci, 13, 98168 Messina, ItalyDepartment of Animal Sciences, Food and Nutrition, Faculty of Agriculture, Food and Environmental Science, Università Cattolica del Sacro Cuore, 29122 Piacenza, ItalyDepartment of Veterinary Sciences, Università di Messina, Viale Palatucci, 13, 98168 Messina, ItalyDepartment of Animal Sciences, Food and Nutrition, Faculty of Agriculture, Food and Environmental Science, Università Cattolica del Sacro Cuore, 29122 Piacenza, ItalyDepartment of Animal Sciences, Food and Nutrition, Faculty of Agriculture, Food and Environmental Science, Università Cattolica del Sacro Cuore, 29122 Piacenza, ItalyDepartment of Veterinary Sciences, Università di Messina, Viale Palatucci, 13, 98168 Messina, ItalyDepartment of Animal Sciences, Food and Nutrition, Faculty of Agriculture, Food and Environmental Science, Università Cattolica del Sacro Cuore, 29122 Piacenza, ItalyDepartment of Veterinary Sciences, Università di Messina, Viale Palatucci, 13, 98168 Messina, ItalyDepartment of Animal Sciences, Food and Nutrition, Faculty of Agriculture, Food and Environmental Science, Università Cattolica del Sacro Cuore, 29122 Piacenza, ItalyAnticoagulants, such as ethylenediaminetetraacetic acid (EDTA), sodium citrate (Na-citrate), or heparin are normally used in hematological clinical tests to prevent coagulation. Although anticoagulants are fundamental for the correct application of clinical tests, they produce adverse effects in different fields, such as those involving specific molecular techniques; for instance, quantitative real time polymerase chain reactions (qPCR) and gene expression evaluation. For this reason, the aim of this study was to evaluate the expression of 14 genes in leukocytes that were isolated from the blood of Holstein cows, and which were collected in Li-heparin, K-EDTA, or Na-citrate tubes; then, they were analyzed using qPCR. Only the <i>SDHA</i> gene showed a significant dependence (<i>p</i> ≤ 0.05) on the anticoagulant that was used with the lowest expression; this was observed in Na-Citrate after being compared with Li-heparin and K-EDTA (<i>p</i> < 0.05). Although a variation in transcript abundance with the three anticoagulants was observed in almost all the investigated genes, the relative abundance levels were not statistically significant. In conclusion, the qPCR results were not influenced by the presence of the anticoagulant; thus, we had the opportunity to choose the test tube that was used in the experiment without interfering effects impacting the gene expression levels caused by the anticoagulant.https://www.mdpi.com/2073-4425/14/2/406HolsteinleukocytesEDTAheparinNa-citrategene expression |
| spellingShingle | Viviana Floridia Marta Sfulcini Enrico D’Alessandro Luca Cattaneo Matteo Mezzetti Luigi Liotta Erminio Trevisi Vincenzo Lopreiato Andrea Minuti Effect of Different Anticoagulant Agents on Immune-Related Genes in Leukocytes Isolated from the Whole-Blood of Holstein Cows Genes Holstein leukocytes EDTA heparin Na-citrate gene expression |
| title | Effect of Different Anticoagulant Agents on Immune-Related Genes in Leukocytes Isolated from the Whole-Blood of Holstein Cows |
| title_full | Effect of Different Anticoagulant Agents on Immune-Related Genes in Leukocytes Isolated from the Whole-Blood of Holstein Cows |
| title_fullStr | Effect of Different Anticoagulant Agents on Immune-Related Genes in Leukocytes Isolated from the Whole-Blood of Holstein Cows |
| title_full_unstemmed | Effect of Different Anticoagulant Agents on Immune-Related Genes in Leukocytes Isolated from the Whole-Blood of Holstein Cows |
| title_short | Effect of Different Anticoagulant Agents on Immune-Related Genes in Leukocytes Isolated from the Whole-Blood of Holstein Cows |
| title_sort | effect of different anticoagulant agents on immune related genes in leukocytes isolated from the whole blood of holstein cows |
| topic | Holstein leukocytes EDTA heparin Na-citrate gene expression |
| url | https://www.mdpi.com/2073-4425/14/2/406 |
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