Activity of GPx in whole blood hemolysates was determined spectrophotometrically with an automated biochemical analyzer RX-Daytona (Randox, Crumlin, UK) using the commercial Ransel kit (Randox Laboratories, Crumlin, UK), which is based on the method of Paglia and Valentine (1967) . Blood hemolysates were diluted 41-fold before analyses with Ransel Diluent (Diluting agent, Randox Laboratories, Crumlin, UK). Activity of GPx was expressed as units per gram of hemoglobin (U/g Hgb). Hemoglobin concentration was determined by the cyanmethemoglobin method using automated hematological analyzer ADVIA 120 (Siemens, Munich, Germany) (Paglia and Valentine, 1967 ). SOD activity in whole blood hemolysates was determined spectrophotometrically with an automatic biochemical analyzer RX Daytona (Randox Laboratories), using commercially available Ransod kit (Randox Laboratories), which is based on the original method of McCord and Fridovich (1969) . Before analyses samples of hemolysates were diluted 1:200 with Ransod Sample Diluent (0.01 mmol/L phosphate buffer, pH 7.0; Randox Laboratories). Activity was expressed as U/g Hgb. Hemoglobin concentration was determined by the cyanmethemoglobin method using automated hematological analyzer ADVIA 120 (Siemens, Munich, Germany) (McCord and Fridovich, 1969 ).
Advia 120
Manufactured by Siemens
Sourced in Germany, United States, France, Japan, Switzerland, United Kingdom, Italy
The Advia 120 is a hematology analyzer designed for clinical laboratory use. It provides automated analysis of blood samples, including red blood cell count, white blood cell count, and platelet count. The Advia 120 is a compact and efficient instrument that can handle a wide range of sample types and deliver accurate, reliable results.
Automatically generated - may contain errors
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Rx daytona, by Randox (7 mentions)
Cobas integra 400 plus, by Roche (4 mentions)
Cobas 6000, by Roche (4 mentions)
Urilux, by Roche (3 mentions)
Sedivue dx, by IDEXX (3 mentions)
Ilyte, by Werfen (3 mentions)
Advia 1650, by Siemens (3 mentions)
Cobas c501, by Roche (3 mentions)
Ransod kit, by Randox (2 mentions)
Ransel kit, by Randox (2 mentions)
Ransel diluent, by Randox (2 mentions)
Ransod sample diluent, by Randox (2 mentions)
Cobas e601, by Roche (2 mentions)
Map k2 edta microcontainer, by BD (2 mentions)
V 6.3.2 ms, by Siemens (2 mentions)
Xe 5000, by Sysmex (2 mentions)
Facscalibur, by BD (2 mentions)
Acl 8000, by Beckman Coulter (2 mentions)
Hystopaque 1077, by Merck Group (2 mentions)
Hemavet cbc analyzer, by Drew Scientific (2 mentions)
144 protocols using advia 120
1
Enzymatic Antioxidant Activities in Whole Blood
2
Plasma Lipid Profile Analysis
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Blood was collected by cardiac puncture from non-fasted, anesthetized animals (see Perfusion method) at harvest prior to incision of the right atrium and subsequent perfusion. A 25-gauge EDTA-coated needle, attached to a 1 ml syringe, is inserted into the right atrium of the exposed heart and the plunger gently pulled to slowly aspirate approximately 500 ml of blood, avoiding entrapping air in the syringe to prevent hemolysis. After removal of the needle from the syringe, the blood was slowly injected into a 1.5 ml EDTA coated MAP-K2 blood microtainer (363706, BD, San Jose, CA) on ice. Blood tubes were spun at 4°C and 4,388× g for 15 min. Blood serum is then removed and aliquoted equally into three replicate 1.5 ml tubes on ice. Tubes were then snap-frozen on dry ice and stored long-term at −80°C. Thawed blood plasma collected from non-fasted mice was then analyzed by Beckman Coulter AU680 chemistry analyzer (Beckman Coulter, Brea, CA) and Siemens Advia 120 (Germany) for levels of non-fasted glucose, total cholesterol, LDL (low-density lipoproteins), HDL (high-density lipoproteins), triglycerides, and NEFA (non-essential fatty acids).
3
Kidney Transplant Recipient Characteristics
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By thorough examination of patients medical records, data about existence and duration of primary chronic kidney disease, arterial hypertension, diabetes mellitus, cardiovascular events as well as time of kidney transplantation (KTX), type and duration of dialysis treatment before KTX were obtained.
Regarding laboratory parameters, all study participants underwent usual peripheral blood sampling, and they were asked to obtain a 24-h urine sample on the same day of the body composition and blood pressure measurement. We collected data on levels of urea (mmol/L), creatinine (mmol/L), uric acid (mmol/L), serum albumin (g/L), phosphates (mmol/L), C-reactive protein (CRP; mg/L), calcium (mmol/L), glucose (mmol/L), triglycerides (mmol/L), total cholesterol (mmol/L), low-density lipoprotein cholesterol (LDL) (mmol/L), haemoglobin (g/L), mean cellular volume (MCV), sodium (mmol/L), potassium (mmol/L), and eGFR using CKD-EPI (mL/min/1.73 m2). A complete blood count was obtained using a hematology analyser (Advia 120, Siemens, Erlangen, Germany) and intact parathyroid hormone (PTH; pmol/L) was measured by immunoassay analyzer (Cobas e601, Roche Diagnostics, Penzberg, Germany).
Regarding laboratory parameters, all study participants underwent usual peripheral blood sampling, and they were asked to obtain a 24-h urine sample on the same day of the body composition and blood pressure measurement. We collected data on levels of urea (mmol/L), creatinine (mmol/L), uric acid (mmol/L), serum albumin (g/L), phosphates (mmol/L), C-reactive protein (CRP; mg/L), calcium (mmol/L), glucose (mmol/L), triglycerides (mmol/L), total cholesterol (mmol/L), low-density lipoprotein cholesterol (LDL) (mmol/L), haemoglobin (g/L), mean cellular volume (MCV), sodium (mmol/L), potassium (mmol/L), and eGFR using CKD-EPI (mL/min/1.73 m2). A complete blood count was obtained using a hematology analyser (Advia 120, Siemens, Erlangen, Germany) and intact parathyroid hormone (PTH; pmol/L) was measured by immunoassay analyzer (Cobas e601, Roche Diagnostics, Penzberg, Germany).
4
Automated Hematological Analysis Protocol
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Blood samples for evaluation of FBC, DIFF and reticulocyte parameters were collected in
K2EDTA tubes (1.5–2.2 mg of dipotassium EDTA dihydrate per
millilitre of blood). The parameters were analysed with the laboratory's
ADVIA® 120 automated haematology analyser (Siemens Healthcare Diagnostics, Inc,
Tarrytown, New York, United States). Cells were counted and sized by light scatter
technology using white light for white cells and laser light for red cells and
platelets. Haemoglobin was measured by the conventional cyanmethaemoglobin method. The
six-part differential analysis was performed in two channels. Cells in the peroxidase
channel were measured by peroxidase staining intensity and light scatter. Cells in the
basophil/lobularity channel were measured by dual laser light scatter, nuclear density
and lobulation index. Reticulocytes were stained with oxazine 750. The films were spread
on the ADVIA® Autoslide slide maker and stained on the HEMA-TEK® 2000
slide stainer (Siemens Healthcare Diagnostics Inc, Tarrytown, New York, United
States).
K2EDTA tubes (1.5–2.2 mg of dipotassium EDTA dihydrate per
millilitre of blood). The parameters were analysed with the laboratory's
ADVIA® 120 automated haematology analyser (Siemens Healthcare Diagnostics, Inc,
Tarrytown, New York, United States). Cells were counted and sized by light scatter
technology using white light for white cells and laser light for red cells and
platelets. Haemoglobin was measured by the conventional cyanmethaemoglobin method. The
six-part differential analysis was performed in two channels. Cells in the peroxidase
channel were measured by peroxidase staining intensity and light scatter. Cells in the
basophil/lobularity channel were measured by dual laser light scatter, nuclear density
and lobulation index. Reticulocytes were stained with oxazine 750. The films were spread
on the ADVIA® Autoslide slide maker and stained on the HEMA-TEK® 2000
slide stainer (Siemens Healthcare Diagnostics Inc, Tarrytown, New York, United
States).
5
Comprehensive Metabolic Biomarker Profiling
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Fasting plasma total cholesterol, triglyceride, glucose, and hemoglobin concentrations were measured at baseline using standard methods as previously described[22 ]: all analyte concentrations were measured on a Modular PP instrument (Roche Diagnostics, Meylan, France), except for hemoglobin which was measured on an ADVIA 120 instrument (Siemens Healthcare Diagnostics, Saint‐Denis, France), at La Conception Hospital (Biochemistry Laboratory, Marseille). All analyses were performed according to manufacturers’ instructions.
Fasting plasma PTF concentration was measured by HPLC as described thereafter.
Fasting plasma PTF concentration was measured by HPLC as described thereafter.
6
Veterinary Hematology Analysis Protocol
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Blood collected in EDTA tubes was analyzed for whole blood count in the Servei d’Hematologia Clínica Veterinària of the UAB (ADVIA 120, Siemens). Laboratory Reference Values were according to previous reports77 .
7
Metabolic Profile and Antioxidant Capacity
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In the whole blood samples, glutathione peroxidase (GSH-Px) was determined with a commercial kit (Ransel test kit, Randox Laboratories Ltd., Crumlin, UK) adapted from the method of Paglia and Valentine [35 (link)]. For the determination of hemoglobin (Hb), a hematological analyzer (Advia 120, Siemens Healthcare Diagnostics SL, Barcelona, Spain) was required. The assessment of the overall metabolic profile in the plasma samples was performed using an automatic chemistry analyzer (Olympus AU600, Olympus Diagnostica Europe GmbH, Ennis, Ireland), and commercial Beckman kits (Beckman Coulter Inc., Fullerton, CA, USA) were used for the assays. Glucose was measured using the hexokinase G-6-PDH method; total triglycerides (TGs) were hydrolyzed using a combination of microbial lipases to produce glycerol and fatty acids; urea was measured using the enzymatic adapted Talke and Schubert [36 (link)] method; total proteins were measured using the adapted Weichselbaum [37 (link)] method; and cholesterol was determined using the cholesterol dehydrogenase method. TAC (antioxidant capacity) and TOS (oxidative status) were analyzed in the plasma samples using the methods described by Erel et al. [38 (link),39 (link)]. T4 total hormone was determined with the immunoassay of chemiluminescence (Immulite 1000 Immunoassay System, Siemens Medical Solutions Diagnostics, Deerfield, IL, USA).
8
Blood Sampling for FACS and Hematology
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Blood was obtained by incision of the tail vein or by puncturing of the heart. For FACS analysis blood was collected in PBS containing 1% heparin (Ratiopharm), followed by erythrocyte depletion as described above. For hematological analysis, blood was collected in EDTA coated tubes (MiniCollect; Greiner) and analyzed by the hematology system ADVIA 120 (Siemens).
9
Immunoglobulin and Hematological Assessments
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The blood samples collected on Days 20 and 29 were centrifuged to obtain serum, and the
concentrations of serum KLH-specific IgM (Days 20 and 29) and IgG (Day 29) were measured
using an ELISA method with an Anti-KLH (TDAR) Rat ELISA KIT (AKRKM-010 for IgM, AKRKG-010
for IgG, Shibayagi, Gunma, Japan).
Hematological parameters including the white blood cell (WBC) count, differential WBC
absolute counts (neutrophils, lymphocytes, monocytes, eosinophils, basophils, and large
unstained cells), red blood cell count, hemoglobin level, hematocrit level, reticulocyte
count, mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH) level, mean
corpuscular hemoglobin concentration (MCHC), and platelet count were measured using a
hematology system (ADVIA 120, Siemens Healthcare Diagnostics K.K., Tokyo, Japan).
concentrations of serum KLH-specific IgM (Days 20 and 29) and IgG (Day 29) were measured
using an ELISA method with an Anti-KLH (TDAR) Rat ELISA KIT (AKRKM-010 for IgM, AKRKG-010
for IgG, Shibayagi, Gunma, Japan).
Hematological parameters including the white blood cell (WBC) count, differential WBC
absolute counts (neutrophils, lymphocytes, monocytes, eosinophils, basophils, and large
unstained cells), red blood cell count, hemoglobin level, hematocrit level, reticulocyte
count, mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH) level, mean
corpuscular hemoglobin concentration (MCHC), and platelet count were measured using a
hematology system (ADVIA 120, Siemens Healthcare Diagnostics K.K., Tokyo, Japan).
10
Hematological and Coagulation Analysis in Ms5Yah Mice
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Blood was collected from 25-week-old males (11 Ms5Yah and 11 wild-type littermates) by retro orbital puncture under isoflurane anaesthesia at 00.00 (time 0) after 4 h of fasting. A total of 120 µl of blood was collected into EDTA-coated paediatric Microvette tubes for haematological analysis. A complete blood count, including total erythrocyte, leukocyte and platelet counts, differential leukocyte count (granulocytes: neutrophils, eosinophils and basophils, lymphocytes and monocytes), haemoglobin and haematocrit measurement, and calculation of blood indexes (mean cell volume, mean corpuscular haemoglobin concentration), was performed on total blood using an Advia 120 haematology analyser (Siemens).
To evaluate whether the platelet deficit of Ms5Yah mice influenced the coagulation process, bleeding time was measured in the same animals at 27 weeks of age. After 5-mm distal tips had been transected, the tail was immersed into PBS at 37°C. Bleeding time was measured from incision to the first time that the bleeding stopped.
To evaluate whether the platelet deficit of Ms5Yah mice influenced the coagulation process, bleeding time was measured in the same animals at 27 weeks of age. After 5-mm distal tips had been transected, the tail was immersed into PBS at 37°C. Bleeding time was measured from incision to the first time that the bleeding stopped.
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