Blood was obtained for each test, from 3 healthy donors (ethical number B03920096633) who were free from any medication for at least two weeks. All protocols were in accordance with the Declaration of Helsinki and approved by the Medical Ethical Committee of the CHU Dinant-Godinne UCL Namur (Yvoir, Belgium). Blood was collected with a 21-gauge needle via atraumatic antecubital venipuncture in 0.109 M sodium citrate (9:1 v/v) tubes Venosafe® (Terumo Europe, Leuven, Belgium) for red blood cells (RBCs) and platelet collection and in EDTA tubes Venosafe® (Terumo Europe, Leuven, Belgium) for white blood cells isolation.
Venosafe
Manufactured by Terumo
Sourced in Belgium
Venosafe is a laboratory blood collection device designed for healthcare professionals. It is used to collect blood samples from patients in a safe and efficient manner.
Automatically generated - may contain errors
Lab products found in correlation
Falcon microtest, by BD (2 mentions)
Human leptin elisa, by BioVendor (2 mentions)
Human ghrelin easy sampling elisa, by BioVendor (2 mentions)
Immulite 2000, by Siemens (2 mentions)
Architect ci8200 analyzer, by Abbott (1 mentions)
Cobas e601, by Roche (1 mentions)
Venosafe plastic tubes, by Terumo (1 mentions)
Konelab 20 xti clinical chemistry analyzer, by Thermo Fisher Scientific (1 mentions)
Konelab 20, by Thermo Fisher Scientific (1 mentions)
12 protocols using venosafe
1
Collection of Blood Samples from Healthy Donors
2
Blood Cytokine Production Assay
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Blood was collected from the caudal blood vessels into commercially available 10-mL evacuated tubes coated with lithium-heparin as anticoagulant (Venosafe™, Terumo® Europe). Samples were used within 4 h after collection. Stimulations were performed in triplicate by mixing in 96-well microplates (Falcon Microtest™, Becton Dickinson) 100 μL of blood with either 100 μL of culture medium (RPMI-1640 supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 10 mM HEPES, penicillin-streptomycin, fungizone and 0.05 mM 2-mercaptoethanol) as negative control, 100 μL of pokeweed mitogen (2 μg/mL) as positive control, or 100 μL of ovalbumin solution (50 μg/mL). The culture was incubated at 37°C in a humidified atmosphere with 5% CO2 for 3 days unless specified otherwise (see Figure legends). Supernatant were then harvested and stored at -20°C in 96-well plastic storage plates (Greiner™ bio-one) until assayed for cytokine content.
3
Venous Blood Sampling for Lipid and Thyroid Assays
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After an overnight fast, venous blood samples for serum lipid assays were drawn into a gel tube containing clot activator and samples for the plasma glucose assay into a fluoride citrate tube (Venosafe; Terumo Europe). The samples were immediately frozen after separating serum and plasma and transferred to the laboratory in dry ice once a week for analyses. The rest of samples were kept in −70°C.
All basic assays were performed at the Laboratory of Analytical Biochemistry at the National Public Health Institute, Helsinki (Disease Risk Unit, Institute for Health and Welfare since 2009), using an ARCHITECT ci8200 analyzer (Abbott Laboratories). High-sensitivity C-reactive protein (hs-CRP) was measured with an ultrasensitive immunoturbidimetric method using Abbott architect reagents.
Free thyroxine (F-T4), free triiodothyronine (F-T3), and thyroid stimulating hormone (TSH) were measured from frozen EDTA-plasma samples with Cobas e601 (Roche Diagnostics) automated analyzer in the Islab Laboratory of the University of Eastern Finland, Kuopio, in the year 2013. The reference values for free T4, free T3, and TSH were 11–22 pmol/L, 3.1–6.8 pmol/L, and 0.3–4.2 mU/L, respectively. Serum thyroid antiperoxidase antibodies were measured by Architect® (Abbott Laboratories) automated analyzer. Analytical sensitivity of the assay was 0.16 IU/mL. The upper reference limit was 6.0 IU/mL.
All basic assays were performed at the Laboratory of Analytical Biochemistry at the National Public Health Institute, Helsinki (Disease Risk Unit, Institute for Health and Welfare since 2009), using an ARCHITECT ci8200 analyzer (Abbott Laboratories). High-sensitivity C-reactive protein (hs-CRP) was measured with an ultrasensitive immunoturbidimetric method using Abbott architect reagents.
Free thyroxine (F-T4), free triiodothyronine (F-T3), and thyroid stimulating hormone (TSH) were measured from frozen EDTA-plasma samples with Cobas e601 (Roche Diagnostics) automated analyzer in the Islab Laboratory of the University of Eastern Finland, Kuopio, in the year 2013. The reference values for free T4, free T3, and TSH were 11–22 pmol/L, 3.1–6.8 pmol/L, and 0.3–4.2 mU/L, respectively. Serum thyroid antiperoxidase antibodies were measured by Architect® (Abbott Laboratories) automated analyzer. Analytical sensitivity of the assay was 0.16 IU/mL. The upper reference limit was 6.0 IU/mL.
4
Minimally Invasive Animal Sampling
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Blood and urine sampling was performed 3–4 weeks before slaughtering, which took place in EU-certified abattoirs. Blood was collected in the morning (09.00–11.00 hours) using 10 ml tubes (Venosafe®, TERUMO) from the jugular vein. After clotting, serum was separated by centrifugation at 1272g for 15 min at RT, divided into aliquots, immediately frozen in liquid nitrogen and then stored at −80°C until analysis. When appropriate, urine samples were concurrently collected after spontaneous micturition, divided into aliquots, immediately frozen in liquid nitrogen and stored at −20°C pending analysis. Particular care was taken at avoiding faecal contamination. Sampling was conducted in the frame of routine controls by licensed veterinarians, avoiding any possible physical pain or stress condition, in accordance with the EU current legislation on animal welfare.
5
Monitoring Chemotherapy Response in Lymphoma
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Blood was collected into 0.109 M sodium citrate (9:1 v/v) tubes (Venosafe;
Terumo, Leuven, Belgium) at patient catheter. Samples were obtained at the
administration of the first chemotherapeutic cure (C0) (just before
administration), at the administration of the second and the fourth cycles of
chemotherapy (C2 and C4), and at the remission review (Cf). In the case of an
autograft, a final sample was taken at the postgraft review. These times were
chosen considering the evaluation of the response to treatment performed by 18 (link)fluorodeoxyglucose positron emission tomography (FDG-PET)/computed
tomography (CT) during the fourth chemotherapeutic cure. The patient response
was evaluated using the Deauville score21 (link) and according to the Lugano classification.22 (link) Platelet-free plasma was obtained following strict pre-analytical setting
already published by our research group.23 (link) Plasma samples were frozen in liquid nitrogen and stored at −80°C.
Terumo, Leuven, Belgium) at patient catheter. Samples were obtained at the
administration of the first chemotherapeutic cure (C0) (just before
administration), at the administration of the second and the fourth cycles of
chemotherapy (C2 and C4), and at the remission review (Cf). In the case of an
autograft, a final sample was taken at the postgraft review. These times were
chosen considering the evaluation of the response to treatment performed by 18 (link)fluorodeoxyglucose positron emission tomography (FDG-PET)/computed
tomography (CT) during the fourth chemotherapeutic cure. The patient response
was evaluated using the Deauville score21 (link) and according to the Lugano classification.22 (link) Platelet-free plasma was obtained following strict pre-analytical setting
already published by our research group.23 (link) Plasma samples were frozen in liquid nitrogen and stored at −80°C.
6
Assessment of Antigen-Specific T Cell Response
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The antigen-specific T cell response was assessed as described [18 (link)]. Blood was collected from the caudal blood vessels into commercially available 10-mL evacuated tubes coated with lithium-heparin as anticoagulant (Venosafe™, Terumo® Europe). Samples were used within 4 h after collection. Stimulations were performed in triplicate by mixing 100 μL of blood with either 100 μL of culture medium (RPMI-1640 supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 10 mM HEPES, penicillin-streptomycin, fungizone and 0.05 mM 2-mercaptoethanol) as negative control, 100 μL of pokeweed mitogen (2 μg/mL) as positive control, or 100 μL of ovalbumin or HEL (50 μg/mL) in 96-well microplates (Falcon Microtest™, Becton Dickinson). The culture was incubated at 37°C in a humidified atmosphere with 5% CO2 for 3 days. Supernatant were then harvested and stored at -20°C in 96-well plastic storage plates (Greiner™ bio-one) until assayed for cytokine content.
7
Metabolic Biomarker Profiling Protocol
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Blood samples for leptin, unacylated ghrelin, creatinine, glucose, urea, free fatty acids and electrolytes (Na2+, K+, Cl–) were drawn from the antecubital vein after overnight fasting. Samples were collected into VenoSafe plastic tubes (VenoSafe®, Terumo Europe, Leuven, Belgium) containing silica gel. Blood samples were centrifuged (3,500 rpm, 10 min) and serum was frozen at −20°C for later analysis. Leptin and ghrelin were determined with an ELISA-kit immunoassay system (Dynex DS 2, Dynex Technologies, Chantilly, VA, United States); creatinine, glucose and urea with a photometric enzymatic method (Konelab 20 Xti Clinical Chemistry Analyzer, Thermo Scientific, Vantaa, Finland); free fatty acids with an enzymatic colorimetric assay (Konelab 20 Xti Clinical Chemistry Analyzer, Thermo Scientific, Vantaa, Finland); and electrolytes (Na, K, Cl) with an ion selective electrode method (ISE; Konelab 20 Xti Clinical Chemistry Analyzer, Thermo Scientific, Vantaa, Finland).
The sensitivity and inter-assay coefficient of variation for these assays were: 0.2 ng/ml, 6.1% for leptin; 0.6 pg/ml, 17.3% for ghrelin; 2.32 μmol/l, 2.2% for creatinine; 0.1 mmol/l; 3.8% for glucose; 1.1 mmol/l, 5.8% for urea, 10 μmol/l, 5.8% for free fatty acids; 100 mmol/l, 0.7% for sodium; 2 mmol/l, 2.5% for potassium and 55 mmol/l, 3.3% for chloride.
The sensitivity and inter-assay coefficient of variation for these assays were: 0.2 ng/ml, 6.1% for leptin; 0.6 pg/ml, 17.3% for ghrelin; 2.32 μmol/l, 2.2% for creatinine; 0.1 mmol/l; 3.8% for glucose; 1.1 mmol/l, 5.8% for urea, 10 μmol/l, 5.8% for free fatty acids; 100 mmol/l, 0.7% for sodium; 2 mmol/l, 2.5% for potassium and 55 mmol/l, 3.3% for chloride.
8
Plasma and Serum Biomarker Analysis
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Blood samples were collected into EDTA tubes (Venosafe®, Terumo, France) at the beginning and the end of the experimental procedure and centrifuged at 1300 g for 10 min at 4°C to separate the plasma which was then rapidly frozen in liquid nitrogen and stored at -80°C until analysis. Blood was also collected into dry tubes (Venosafe®, Terumo, France) following depletion period, and after an incubation for 20 min at room temperature they were centrifuged at 1300 g for 10 min at 4°C to separate the serum which was then rapidly frozen in liquid nitrogen and stored at -80°C until analysis.
Plasma 25(OH) D levels were measured using a 25-OH Vitamin D (direct) ELISA kit (PromoKine, France) according to the manufacturer’s instructions.
Serum calcium and phosphorus levels were measured using an automat Konelab 20 (Thermo Scientific, MA, United States).
Plasma 25(OH) D levels were measured using a 25-OH Vitamin D (direct) ELISA kit (PromoKine, France) according to the manufacturer’s instructions.
Serum calcium and phosphorus levels were measured using an automat Konelab 20 (Thermo Scientific, MA, United States).
9
PRP Preparation for Dasatinib Patients
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Platelet-rich plasma (PRP) was prepared from blood of patients treated with dasatinib and from blood of healthy volunteer, matched for age and sex, not taking any drugs in the previous 3 weeks.
Blood was taken 3 and 24 h after drug administration, corresponding to the expected peak of dasatinib in peripheral blood. Concerning the patients characteristics, the mean time from diagnosis of chronic myeloid leukemia (CML) in chronic phase was 25,5 months (range 6–60). All the patients discontinued treatment with imatinib due to adverse reaction or treatment failure and were on treatment with dasatinib (100 mg/day in four patients, 140 mg/day in one patient) from 9 months (range 2–32). In all subjects blood was drawn by venepuncture into 3,6 ml vacutainer (Venosafe, Terumo) with trisodium citrate 0.109M . To block platelets cyclooxygenase activity ASA 100 μM was added to blood samples. For western blot experiments blood was collected in 3 ml Hirudine Blood Tube (Verum Diagnostica) in the presence of trisodium citrate 3.8%.
Platelet-rich plasma was obtained by centrifugation of blood at 200 × g at room temperature for 10 min and platelet count was estimated by an automated cell counter.
Blood was taken 3 and 24 h after drug administration, corresponding to the expected peak of dasatinib in peripheral blood. Concerning the patients characteristics, the mean time from diagnosis of chronic myeloid leukemia (CML) in chronic phase was 25,5 months (range 6–60). All the patients discontinued treatment with imatinib due to adverse reaction or treatment failure and were on treatment with dasatinib (100 mg/day in four patients, 140 mg/day in one patient) from 9 months (range 2–32). In all subjects blood was drawn by venepuncture into 3,6 ml vacutainer (Venosafe, Terumo) with trisodium citrate 0.109
Platelet-rich plasma was obtained by centrifugation of blood at 200 × g at room temperature for 10 min and platelet count was estimated by an automated cell counter.
10
Fasting Blood Hormone Analysis
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Blood samples were collected in the morning (7:00–9:00 a.m.) after a 12 h overnight fast. Participants were instructed to abstain from strenuous physical activity for 24 h before the blood samples were taken. Venous blood samples were drawn from an antecubital vein using standard procedures and the blood was transferred into serum and EDTA tubes (Venosafe, Terumo, Belgium). The serum samples were held for 15 min at room temperature before being centrifuged for 10 min at 2000× g (Megafuge 1.0 R, Heraeus, Germany). The serum was separated and immediately frozen at −80 °C for later analysis. Leptin was assessed with the Biovendor Human Leptin ELISA. Total ghrelin was assessed with the Biovendor Human Ghrelin Easy Sampling ELISA from plasma after incubation at room temperature for 2 h. The assay sensitivity for ghrelin was 10 mg·L−1. Other hormonal analyses were performed using chemical luminescence techniques (Immulite 2000, Siemens Healthcare Diagnostics, Camberley, UK) with an assay sensitivity of 55.0 pmol·L−1 for E2, 0.3 ng·mL−1 for P4, 0.2 ng·mL−1 for leptin, 1.5 mmol·L−1 for T3, 10 ng·L−1 for ghrelin, 2 mIU·l−1 for insulin, and 0.1 nmol·L−1 for glucose. Inter-assay coefficients of variation (CV) were 6.7% for E2, 9.7% for P4, 4.2% for leptin, 8.1% for T3, 6.8% for ghrelin, 5.1% for insulin, and 1.4% for glucose.
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