Blood samples were aseptically collected for haematology and clinical biochemistry from the distal cephalic vein and transferred into EDTA and non-additive collection tubes (Vacutainer®, Becton-Dickinson, Stockholm, Sweden). The centrifugation was performed by using the blood collected in the non-additive tubes. The separated serum was used for analysing clinical biochemistry parameters. Clinical biochemical included Bile acids, Alanine aminotransferase (ALT), Glucose, Blood urea nitrogen (BUN), and Creatinine were measured (Abbott Architect c4000, Abbott Park, IL, USA). Haematological analyses (WBC including differential counts, haematocrit (PCV) and haemoglobin (Hb)) were performed (Advia 2120; Siemens Healthcare Diagnostics, Deer-field, IL, USA). Colorimetric method (bromocresol green) was used to analyse albumin and measured by using an automated analyser (Abbott Architect c4000, Abbott Park, IL, USA). All laboratory analyses were performed according to the routine methods at the Clinical Pathology Laboratory, UDS, SLU, Uppsala, Sweden.
Architect c4000
Manufactured by Abbott
Sourced in United States
The Architect c4000 is a high-throughput, fully automated immunoassay analyzer designed for clinical laboratory testing. It is capable of performing a variety of immunoassay tests, including those for infectious diseases, hormones, and other analytes.
Automatically generated - may contain errors
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48 protocols using architect c4000
1
Comprehensive Blood Analysis Protocol
2
Urine and Serum Analyses in Canine Babesiosis
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Voided midstream urine samples from healthy dogs and dogs with babesiosis were collected into sterile tubes. After urinalysis, the remaining urine was centrifuged at 500 g for 10 min at 4°C, and the supernatant was aliquoted and stored at −80°C until analysis. Routine urinalysis including semiquantitative dipstick test (Multistix 10SG, Siemens) was performed on an automated analyser (CLINITEK Status + Analyzer, Bayer) and the microscopic sediment analysis at a high (400x) power field. Urine protein and creatinine concentrations were determined using commercial kits on an automated chemistry analyser (Architect c4000, Abbot, IL, United States).
Serum samples were also collected for biochemistry analyses performed on the biochemistry analyser (Architect c4000, Abbot, IL, United States), while C-reactive protein (CRP) was assessed by canine specific assay (Gentian Diagnostics ASA, Moss, Norway) on the same analyser. Hematological data was generated from the plasma samples using an automatic hematology analyser, Horiba ABX (Diagnostics, Montpellier, France).
Serum samples were also collected for biochemistry analyses performed on the biochemistry analyser (Architect c4000, Abbot, IL, United States), while C-reactive protein (CRP) was assessed by canine specific assay (Gentian Diagnostics ASA, Moss, Norway) on the same analyser. Hematological data was generated from the plasma samples using an automatic hematology analyser, Horiba ABX (Diagnostics, Montpellier, France).
3
Plasma Glucose Analysis Post-Decapitation
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After decapitation, trunk blood was collected at the decapitation site in heparinized tubes before centrifugation at 1000 ×g for 10 min. The plasma was collected and stored at −80 °C until analysis. Glucose levels were analyzed with enzymatic colorimetric methods using an Architect c4000 automated chemistry analyzer (Abbott Diagnostics, Lake Forest, IL,USA).
4
Impaired Glucose Tolerance Diagnostic Protocol
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Study participants were recruited at the Polyclinic Department of the Endocrinology Research Centre (Moscow, Russia). The study was approved by the ethical review committee #27-01 of the RAMS (Moscow, Russia), approval number #64 (statement # 01-02/62). Subjects at risk for diabetes who were admitted to the department were selected to participate in this study. All participants signed their written informed consent to provide blood samples for research purposes. Blood plasma concentrations of diagnostic substances (glucose, uric acid, total cholesterol, insulin, triglycerides, low-density lipoprotein (LDL), and high-density lipoprotein (HDL) were measured using the Architect c4000 clinical chemistry analyzer (Abbott Diagnostics, Abbott Park, IL, USA). Glycated hemoglobin (HbA1c) was measured using the Bio-Rad D10 hemoglobin testing system (Bio-Rad Laboratories, France). For the oral glucose talerance test (OGTT), a standard glucose dose (75 g) was orally ingested and blood glucose levels were checked two hours later. IGT was diagnosed if the post-load glucose levels were between 7.8 and 11.0 mmol/l (W.H.O. 1999) [18] . In this study, OGTT results were used to establish gender-matched cases (IGT; n = 20) and control (Normal; n = 30) groups. Table 1 (and Table S1 ) presents the clinical characteristics of the cohort.
5
Metabolic Biomarkers Assessment Protocol
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HbA1c (reference values 4%–6%) was assessed by high-performance liquid chromatography (D10 Hemoglobin Testing System, Bio-Rad, France). Fasting blood glucose (FBG) (fasting reference values 3.1–6.1 mM) was assessed by ARCHITECT c4000 Clinical Chemistry Analyzer (Abbott Diagnostics, Abbott Park, IL, USA) with manufacturer kits. Immune-reactive insulin was measured in serum with standard kit using electrochemiluminescence analyzer Cobas 6000 (Roche, Switzerland). ELISA kits for adiponectin, leptin, glucagon, and GLP-1 were obtained from Mercodia (Sweden), for GIP—from Cloud-Clone Corp. (USA), and for oxyntomodulin—from Cusabio (USA). The ELISA measurements were performed using 1420 Multilabel Counter VICTOR2 (PerkinElmer, USA).
6
Hormonal and Metabolic Profiling of Donors
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Two hormones were measured in each donor, basal insulin and serum cortisol, as well as serum glucose concentration. A blood sample was collected by puncture of the coccygeal vein, one day before each OPU at 6:00 h, with 21G × 38 mm needles and Vacutainer tubes (Vacutainer ® , Becton Dickinson, Franklin Lakes, NJ, USA). Blood samples were collected and centrifuged for 15 min at 750× g at 4 • C. Serum samples were frozen and kept at -20 • C until further analysis. Basal insulin and serum cortisol concentrations were determined by chemiluminescence immunoassay (CLIA) (Maglumi 800; Shenzhen New Industries Biomedical Engineering Co., Ltd. (Snibe), Shenzhen, China), using commercial kits MAGLUMI ® insulin (CLIA, Shenzhen, China) and MAGLUMI ® cortisol (CLIA, Shenzhen, China), with intra-and inter-assay coefficient of variation (CV) of 36.9 and 37.4 to insulin and 77.7 and 81.1 to cortisol, respectively. Serum glucose concentration was determined by automated spectrophotometry with a clinical chemistry analyzer (Architect c4000; Abbott Diagnostics, Abbott Park, IL, USA) using a commercial reagent 3L82-20 with intra-and inter-assay CV of 9.4 and 9.3, respectively.
7
Plasma Glucose Analysis Post-Decapitation
8
Oral Glucose Tolerance Test Protocol
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The oral glucose tolerance test was performed according to the recommendations of the WHO. The glucose load was 1.75 g of glucose per kg body weight up to a total of 75 g of glucose. Blood samples were obtained at 0 and 120 minutes for the measurement of glucose. [19] Laboratory analysis Venous blood samples were collected following standard procedures from the antecubital vein after overnight fasting of 10-12 hours. The samples were analyzed for concentrations of: plasma glucose, total serum cholesterol, serum triglycerides, serum alanine amino trasnferase (ALT) and serum aspartate aminotransferase (AST). Two hours after the performance of OGTT, blood samples were obtained and plasma glucose concentration was determined. All blood samples were analyzed in clinical chemistry analyzer AR-CHITECT c4000 (Abbott Diagnostics).
9
Fasting Lipid Biomarker Measurement
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Blood samples were collected after an overnight fast. Standard commercially-available assays adapted to an Architect C4000 (Abbott Diagnostics, USA) were used to determine plasma total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-c) and apoB. LDL-c was calculated by the Friedewald-formula [10] , since all TG were < 300 mg/dl.
10
Paraoxonase 1 and Albumin in CSF
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Frozen aliquots of CSF (IVDH N = 5 and control N = 6) were thawed to determine paraoxonase 1 (PON1) activity, as well as albumin (ALB) concentration using automated biochemistry analyzer Abbott Architect c4000 (Abbott, Chicago, IL, USA). PON1 activity was assayed using the method of Tvarijonaviciute et al. with p-nitrophenyl acetate (Sigma-Aldrich, Saint Louis, MO, USA) as substrate [74 (link)]. Albumin concentration in CSF samples was evaluated with human immunoturbidimetric assay (Microalbumin OSR6167 Beckman Coulter, Brea, CA, USA) which was previously validated for use in canine CSF samples [75 (link)].
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