A fasting blood sample (7 mL) was taken from each participant in the morning (7:00‐8:30 am ) and collected in vacuum tubes (Becton Dickinson, Franklin Lakes, NJ, USA). Samples were centrifuged at 250 g rpm for 15 minutes (Beckman Allegra 21R, Palo Alto, CA, USA), and plasma was separated. Concentrations of leptin and C‐Reactive Protein (CRP) in plasma were measured in duplicate using commercial ELISA kits (Human Leptin Elisa Kit, Linco Research; High‐Sensitivity C‐Reactive Protein ELISA Kit, Bioquant). The concentration of the inflammatory cytokines TNF‐α, IL‐6 and the regulatory cytokine IL‐10 was measured using high‐sensitivity commercial ELISA kits (Millipore CRP ELISA, MO, USA). All ELISA kits were analysed in a Multiskan Ascent microplate photometer (Thermo Electron Corporation, MA, USA). All the biochemical analyses were performed by trained personnel at the Human Nutrition Laboratory, UAQ.
Vacuum tubes
Manufactured by BD
Sourced in United States, France
Vacuum tubes are electronic devices that operate by the flow of electrons in a vacuum or partial vacuum. They are used in various electronic circuits and devices to amplify, control, or generate electrical signals.
Automatically generated - may contain errors
Lab products found in correlation
Allegra 21r, by Beckman Coulter (1 mentions)
Human leptin elisa kit, by Linco Research (1 mentions)
Kx 21, by Sysmex (1 mentions)
Nunc polypropylene cryotubes, by Thermo Fisher Scientific (1 mentions)
Metrolab 2300 auto analyzer, by Wiener Lab (1 mentions)
Unicel dxi 800, by Beckman Coulter (1 mentions)
Au5800, by Beckman Coulter (1 mentions)
Infinite m200, by Tecan (1 mentions)
Minimacs columns, by Miltenyi Biotec (1 mentions)
Myeloid derived suppressor cell isolation kit, by Miltenyi Biotec (1 mentions)
Ethylene diamine tetraacetic acid (edta), by BD (1 mentions)
Sodium heparin, by BD (1 mentions)
7020 automatic blood analyzer, by Hitachi (1 mentions)
Colibri, by Berthold Technologies (1 mentions)
Qiaamp dna mini kit, by Qiagen (1 mentions)
23 protocols using vacuum tubes
1
Cytokine and Inflammatory Biomarkers in Fasting Plasma
2
Biomarker Profiling of Participants
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Blood samples were collected from the participants by venipuncture in vacuum tubes (Beckton-Dickinson, Mexico City, Mexico), with a previous 8 h fast to determine glucose, HDL cholesterol and triglycerides, urea, uric acid, albumin, and C-reactive protein. In all cases, commercial kits (Randox Laboratories Ltd., Crumlin, London, UK) were used.
3
Platelet Function Analysis Protocol
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Blood samples for the platelet function analysis were collected at 0 (pre-dose), 0.5, 1, 2, 4, 8, 24, 48, and 72 h after being loaded into 2 vacuum tubes (Becton-Dickinson, Franklin Lakes, NJ, USA) containing 3.2% trisodium citrate. The first tube was discarded to avoid spontaneous platelet activation. The second tube was gently inverted 3 times to ensure complete mixing of the anticoagulant and blood sample and then left to stand for at least 10 minutes at room temperature before analysis. The blood sample was analyzed within 2 h for the rapid platelet-function assay.
Platelet activity was analyzed using VerifyNow P2Y12 (Accumetrics, San Diego, CA, USA). The results are reported in P2Y12 reaction units (PRUs), baseline values (BASEs), and percentage inhibition. The percentage inhibition was calculated as follows: ([BASE-PRU]/BASE) × 100. A PRU > 208 was defined as high on-treatment platelet reactivity (HTPR), which is related to a higher risk of an ischemic event; a PRU < 85 was defined as low on-treatment platelet reactivity, which is related to a higher risk of a bleeding event. The technical details of the platelet function assay have been previously described17 .
Platelet activity was analyzed using VerifyNow P2Y12 (Accumetrics, San Diego, CA, USA). The results are reported in P2Y12 reaction units (PRUs), baseline values (BASEs), and percentage inhibition. The percentage inhibition was calculated as follows: ([BASE-PRU]/BASE) × 100. A PRU > 208 was defined as high on-treatment platelet reactivity (HTPR), which is related to a higher risk of an ischemic event; a PRU < 85 was defined as low on-treatment platelet reactivity, which is related to a higher risk of a bleeding event. The technical details of the platelet function assay have been previously described17 .
4
Thrombin Generation Assay for Coagulation
5
Piglet Blood and Fecal Sampling
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On d 14 and 28, blood samples (5 mL) from 1 piglet per pen were collected from the jugular veins using vacuum tubes (Becton–Dickinson Vacutainer Systems, Franklin Lakes, NJ, USA). All samples were centrifuged at 3,000 × g for 10 min at 4 °C, and then stored at −20 °C. During d 26 to 28, feces (200 g) were collected from each pen and then dried at 65 °C for 72 h. On d 28, 6 fresh fecal samples (one piglet per pen) from each treatment were rapidly frozen in liquid nitrogen, and then stored at −80 °C.
6
Plasma Biomarker Sampling Protocol
7
Rat Blood Hematological Analysis
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At the end of the experiment, two weaned rats from each repetition were chosen for blood collection from the canthus of eye, and the rats were anesthetized with isoflurane inhalation first. The blood samples were transferred to the vacuum tubes (Becton Dickinson, Franklin Lakes, NJ, USA) treated with ethylenediaminetetraacetic acid dipotassium anticoagulant (EDTAK2) and stored at 4 °C. The blood samples were gently shaken, and the number of red blood cells (RBCs), hemoglobin (HGB) concentration, and hematocrit (HCT) level were determined by an automatic hematology analyzer (Sysmex KX-21, Sysmex Corporation, Kobe, Japan).
8
Serum Collection and Cryopreservation
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Venous blood samples (10 mL) were collected in vacuum tubes without additives (Becton Dickinson and Company, Franklin Lakes, NJ, USA) and coagulated for 45 min before centrifugation (2000 g for 15 min). Serum was aliquoted into 4.0 mL NUNC® polypropylene cryotubes (Thermo Fisher Scientific, Waltham, MA, USA), and thereafter frozen before long-term storage at -80 °C at the National Institute of Occupational Health (Oslo, Norway).
9
Quantification of Platelet-Derived Proteins
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Before clinical treatment peripheral blood were obtained in vacuum tubes (Becton Dickinson and Company, USA) in the morning before breakfast and centrifuged at 1500 g for 15 min. The supernatants were stored at 80°C until assayed. PDPN and GPVI concentrations were measured using commercially available enzyme-linked immunosorbent assay (ELISA) kits according to the manufacturer's instruction (Cloud-Clone Corp. Houston, USA). CLEC-2 concentrations were measured using commercially available ELISA kits according to the manufacturer's instruction (RayBiotech, Inc.Georgia, USA). All samples were analyzed in duplicate. The results were expressed as nanograms per milliliter. The low limits of detection for PDPN, GPVI and CLEC-2 were 0.156 ng/ml, 0.0156 ng/ml and 1.229 ng/ml respectively.
10
Lipopolysaccharide-Induced Inflammatory Response
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Eighteen hours prior to the postmortem exam, NS and AS groups were injected with a 0.9% physiological saline solution (0.5 mg/kg) and AL, AL-C5, AL-C10 groups were injected with LPS (0.5 mg/kg body weight) into their abdominal cavity. Following a 12-h fast on the last day of the 6-week experimental period, blood was drawn from the saphenous vein of etherized rats. Blood samples were collected in vacuum tubes (Becton Dickinson, Meylan, France) without anticoagulant for biochemical analysis and with heparin for erythrocyte and plasma analyses.
All blood samples were immediately centrifuged at 3,000 rpm (4°C) for 15 min (Sovall ST16R Centrifuge, Thermo Scientific Co., Rockford, IL, USA) to separate serum, plasma, and erythrocytes. Erythrocyte samples were lysed in 4 volumes of ice cold HPLC-grade water, centrifuged (1,000 g, 15 min, 4°C), and the supernatant (erythrocyte lystate) was collected for later analysis. The liver was rinsed with physiological saline after blood collection, exterior moisture was removed, weight was measured, and the sample was frozen in liquid nitrogen for storage until analysis.
All blood samples were immediately centrifuged at 3,000 rpm (4°C) for 15 min (Sovall ST16R Centrifuge, Thermo Scientific Co., Rockford, IL, USA) to separate serum, plasma, and erythrocytes. Erythrocyte samples were lysed in 4 volumes of ice cold HPLC-grade water, centrifuged (1,000 g, 15 min, 4°C), and the supernatant (erythrocyte lystate) was collected for later analysis. The liver was rinsed with physiological saline after blood collection, exterior moisture was removed, weight was measured, and the sample was frozen in liquid nitrogen for storage until analysis.
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