Five biochemical parameters, aspartate aminotransferase (AST), alanine aminotransferase (ALT), gamma glutamyltranspeptidase (GGT), alkaline phosphatase (ALP) and total bilirubin were estimated, as reflection of liver function, by reported method (Edwards and Bouchier, 1991 ). The enzyme activities were measured using diagnostic strips (Reflotron®, ROCHE) and were read on a Reflotron®Plus instrument (ROCHE). TG (Foster and Dunn, 1973 ), TC (Zlatkis et al., 1953 (link)), HDL-C (Burstein et al., 1970 (link)), LDL-C and VLDL (Friedwald et al., 1972 (link)) were assessed using Roche Diagnostics Kits.
Diagnostics kit
Manufactured by Roche
Sourced in Switzerland, Germany, Japan
Roche Diagnostics kits are laboratory equipment designed for clinical diagnostic testing. The kits contain reagents, consumables, and instruments necessary for various analytical procedures. They are intended to aid healthcare professionals in the identification, monitoring, and management of medical conditions.
Automatically generated - may contain errors
Lab products found in correlation
Reflotron plus instrument, by Roche (1 mentions)
Cobas 8000 instrument, by Roche (1 mentions)
Dxh 800 hematology analyzer, by Beckman Coulter (1 mentions)
Vacuette blood collection tube, by Greiner (1 mentions)
Cobas e411, by Roche (1 mentions)
Access 2 immunoassay system, by Beckman Coulter (1 mentions)
Tunel reagent, by Roche (1 mentions)
16 protocols using diagnostics kit
1
Liver Function and Lipid Profile Evaluation
2
Quantifying Osteogenic Differentiation of BMSCs
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Alkaline phosphatase (ALP) and osteocalcin (OCN) activity of the BMSCs were detected using Roche Diagnostics kits (Mannheim, Germany). The culture was similar as described in Section 2.7.1 , but sampled at different time points. ALP activity in medium was measured on Days 3, 7 and 14. The OCN activity was measured using N-MID Osteocalcin kits (Mannheim, Germany).
After the BMSCs were cultured for 21 days, they were washed three times with PBS, fixed and stained with Alizarin Red. The cell monolayers were washed with distilled water until no color appeared, and the images were photographed. The deposits were extracted using 10% (w/v) cetylpyridinium chloride in 10 mM sodium phosphate (pH 7.0) for 2 hours at room temperature to quantify the amount of Alizarin Red. The content of Alizarin Red was determined by its optical density (OD) (Tang et al., 2016 (link)).
After the BMSCs were cultured for 21 days, they were washed three times with PBS, fixed and stained with Alizarin Red. The cell monolayers were washed with distilled water until no color appeared, and the images were photographed. The deposits were extracted using 10% (w/v) cetylpyridinium chloride in 10 mM sodium phosphate (pH 7.0) for 2 hours at room temperature to quantify the amount of Alizarin Red. The content of Alizarin Red was determined by its optical density (OD) (Tang et al., 2016 (link)).
3
Vitamin D, PTH, and Bone Biomarker Analysis
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Total vitamin D (25(OH)D3 and 25(OH)D2) and PTH were measured by chemiluminescent microparticle immunoassay and calcium and phosphate by spectrophotometry, using Roche Diagnostics kits (Switzerland) in our routine clinical laboratory. Accuracy and reliability of this immunoassay for vitamin D were assessed with a standardization program and in external quality assessment scheme.
Various cutoffs to stratify vitamin D levels exist. Following recent consensuses, we defined deficiency as a 25(OH)D concentration of ≤ 50 nmol/L (20 ng/ml) and severe deficiency as < 25 nmol/L (10 ng/ml) [16 (link), 19 (link)]. These thresholds are based on bone outcomes with increased PTH below 50 nmol/L and risk of rickets/osteomalacia below 25 nmol/L [1 (link)]. Serum calcium was considered normal between 2.2 and 2.52 nmol/L and PTH between 1.1 and 6.8 pmol/L. Phosphate thresholds were adapted to age according to the CALIPER program [20 (link), 21 (link)].
Various cutoffs to stratify vitamin D levels exist. Following recent consensuses, we defined deficiency as a 25(OH)D concentration of ≤ 50 nmol/L (20 ng/ml) and severe deficiency as < 25 nmol/L (10 ng/ml) [16 (link), 19 (link)]. These thresholds are based on bone outcomes with increased PTH below 50 nmol/L and risk of rickets/osteomalacia below 25 nmol/L [1 (link)]. Serum calcium was considered normal between 2.2 and 2.52 nmol/L and PTH between 1.1 and 6.8 pmol/L. Phosphate thresholds were adapted to age according to the CALIPER program [20 (link), 21 (link)].
4
Pediatric Venous Blood Collection and Analysis
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Non-fasting blood samples of 10 mL peripheral venous blood were obtained from each child by a trained pediatrician. Blood was drawn into plain and EDTA-containing VACUETTE® Blood Collection Tubes (Greiner Bio one GmbH, Kremsmünster, Austria) handled, and stored by trained staff according to accepted protocols. The blood samples were transferred in cool conditions to a central clinical diagnostic laboratory at Hillel Yaffe Medical Center within 2 h of collection. All assays were performed on the same day in a blinded manner to the demographic, clinical, and nutritional data. A complete blood count (CBC) was conducted using a DxH-800 hematology analyzer (Beckman Coulter Inc., Brea, CA, USA). Levels of biochemical markers in serum were measured using Cobas-8000 instrument with Roche Diagnostics kits according to the manufacturer’s procedures (Roche Diagnostics GmbH, Mannheim, Germany).
5
Serum Lipid Profile Estimation
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The estimation of serum-free fatty acids (FFAs), total cholesterol (TC), phospholipids (PLs) and triglycerides (TGs), high density lipoproteins (HDLs), low-density lipoproteins (LDLs), and very low-density lipoprotein (VLDL) estimations were carried out by Roche diagnostics kits (Roche Diagnostics GmbH, Mannheim, Germany) or commercially available kits from Merck India.
6
Serum Biomarkers in Aging Poultry
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At 78 weeks of age, two birds were selected from each cage (16 birds/group) to collect 3 mL blood samples via the brachial vein route. In order to separate sera, blood samples were centrifuged as quickly as possible (1008× g) and then stored at −20 °C. The concentrations of serum creatine and alanine aminotransferase (ALT) were with commercial Roche diagnostics kits (GmbH, Mannheim, Germany).
7
Lipid Biomarker Analysis in Intervention Study
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Blood samples were collected after twelve-hour overnight fasting at the beginning, and after every 2 weeks, of intervention. The final samples were taken after 10 weeks of intervention. Serum from blood was obtained through separation by centrifugation at 3500 rpm (10 min) and stored at -80°C. In all cases, aliquots of samples were analyzed on the same day as the entire experiment was completed. Concentrations of plasma TG and TC and LDL-C were analyzed using Roche Diagnostics kits (Basel, Switzerland; Ref. No. for kits: TG -11730711, TC -11491458, LDL-C -27714423).
The HDL-C level was determined after separation of apolipoprotein B-containing particles using a phosphotungstate-based method. The Hitachi 920 autoanalyzer (Hitachi, Tokyo, Japan) was used for all measurements. The laboratory was under the External Quality Assurance program of the Centers for Disease Prevention and Control (CDC; Atlanta, USA).
The HDL-C level was determined after separation of apolipoprotein B-containing particles using a phosphotungstate-based method. The Hitachi 920 autoanalyzer (Hitachi, Tokyo, Japan) was used for all measurements. The laboratory was under the External Quality Assurance program of the Centers for Disease Prevention and Control (CDC; Atlanta, USA).
8
Serum Lipid Profile Determination
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Serum total cholesterol (TC), high-density lipoprotein cholesterol (HDL), and triglycerides (TG) were determined by the Roche diagnostics kits (Roche Diagnostics GmbH, Mannheim, Germany) using the previously reported protocols. The levels of low-density lipoprotein cholesterol (LDL) and very low-density lipoprotein cholesterol (VLDL) were computed using the following equations: LDLC = TC-HDL-VLDL; VLDL = TG/5.27.
9
Femoral Vein Blood Collection and Serum Analysis
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Approximately 1.5 ml blood were collected from the femoral vein via needle and syringe and transferred to a serum collection tube. After centrifugation (3,000 rpm for 10 min), approximately 200 μl of serum were transferred into a cryogen tube and stored frozen at approximately À20 C or below. The serum was used for the analyses of all clinical chemistry parameters. When necessary, serum was stored refrigerated up to 3 days at 2 to 8 C until analysis (e.g., CK isoenzymes). Automated methodologies for clinical chemistry were employed (Hitachi 917; Software version: # 02-49; Boehringer Mannheim/Hitachi 917 Operator's Manual). Reagents were Roche Diagnostics kits (Roche Diagnostics; Indianapolis, IN).
Troponin I and Troponin T tests were performed at Ani Lytics, Inc. (Gaithersburg, MD), using an electrochemiluminescent immunoassay (BioVeris, Inc., Gaithersburg, MD)
Troponin I and Troponin T tests were performed at Ani Lytics, Inc. (Gaithersburg, MD), using an electrochemiluminescent immunoassay (BioVeris, Inc., Gaithersburg, MD)
10
Cobas e411 Clinical Chemistry Analysis
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The biochemical parameters were measured using a Cobas e411 clinical chemistry analyzer (Roche Diagnostics, Basel, Switzerland) and using Roche Diagnostics Kits according to the manufacturer's instructions.
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