All the cases of pregnancy toxemia were diagnosed using Rothera’s qualitative test from urine samples [5 (link)]. The blood samples were collected from jugular vein in 20 healthy pregnant as control and 45 pregnancy toxemic goats to carry out different laboratory and biochemical tests by standard kits. Serum glutamic pyruvic transaminase (SGPT) and serum glutamic-oxaloacetic transaminase (SGOT) were estimated by modified IFCC method (Coral Pvt. Ltd). Glucose, blood urea nitrogen (BUN), calcium, magnesium, phosphorus, and creatinine were estimated by glucose oxidase-peroxidase, diacetylmonoxime, o-cresolphthalein complexone, calmagite, Molybdate U.V., and Mod. Zaffe’s Kinetic method. β-hydroxyl butyric acid (BHBA) was estimated by D-3-hydroxybutyrate kit and non-esterified fatty acid (NEFA) were estimated by NEFA Kit (Randox Laboratories Ltd).
Nefa kit
Manufactured by Randox
Sourced in United Kingdom
The NEFA kit is a laboratory tool designed to quantitatively measure the levels of non-esterified fatty acids (NEFA) in various biological samples. It provides a reliable and efficient method for NEFA analysis.
Automatically generated - may contain errors
Lab products found in correlation
Rx daytona, by Randox (2 mentions)
Glycerol kit, by Randox (2 mentions)
Galactose assay kit, by Merck Group (2 mentions)
Bovine igg1 elisa quantitation set, by Fortis Life Sciences (1 mentions)
Biotek synergy plate reader, by Agilent Technologies (1 mentions)
Colorimetric assay, by Merck Group (1 mentions)
Ultrasensitive insulin elisa, by Mercodia (1 mentions)
Mbs703041, by MyBioSource (1 mentions)
Agilent 6470, by Agilent Technologies (1 mentions)
Enzyme linked immunosorbent assay, by Merck Group (1 mentions)
Rat mouse insulin kit, by Merck Group (1 mentions)
Abx pentra 400, by Horiba (1 mentions)
Multiskan spectrum, by Thermo Fisher Scientific (1 mentions)
Mouse rat leptin quantikine elisa kit, by R&D Systems (1 mentions)
11 protocols using nefa kit
1
Pregnancy Toxemia Biochemical Markers
2
Plasma Analyte Concentration Quantification
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The plasma glucose and nonesterified fatty acids (NEFA) concentrations were determined using glucose kit and NEFA kit (Randox Laboratories Ltd, UK), respectively. An automated RX series Clinical Chemistry Analysers (Randox Laboratories Ltd, UK) was used for all measurements. The plasma IgG1 concentrations were detected using a bovine IgG1 ELISA Quantitation set (Bethyl Laboratories, USA) according to the manufacturer’s protocol. Plasma was diluted in Tris-buffered saline (TBS)-Tween to a final dilution factor of 4 × 104. All dilutions were duplicated. Absorbance was read using a BioTek Synergy plate reader (BioTek Instruments, Inc., USA) at a wavelength of 450 nm.
3
Plasma Biomarker Analysis Protocol
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Venous blood samples were centrifuged at 1865g for 15 min at 4°C. Aliquots of plasma were immediately frozen and stored at −70°C for later analysis. Plasma was analyzed using commercially available kits for glucose, lactate, and nonesterified fatty acid (NEFA) concentrations (Glucose kit, Lactate kit, NEFA kit; Randox, London, United Kingdom) using an automated photometric clinical chemistry analyzer RX Daytona+ (Randox). Plasma galactose concentration was quantified using a colorimetric assay (Sigma Aldrich, St Louis. MO) and insulin using an enzyme-linked immunosorbent assay (Ultrasensitive Insulin ELISA; Mercodia, Uppsala, Sweden).
Isotopic enrichment of breath samples was determined using gas chromatography isotope ratio mass spectrometry (Europa Scientific Hydra 20-20, Crewe, United Kingdom). 13C enrichment of ingested CHOs was determined by elemental analyzer isotope ratio mass spectrometry (Europa Scientific Hydra 20-20).
Isotopic enrichment of breath samples was determined using gas chromatography isotope ratio mass spectrometry (Europa Scientific Hydra 20-20, Crewe, United Kingdom). 13C enrichment of ingested CHOs was determined by elemental analyzer isotope ratio mass spectrometry (Europa Scientific Hydra 20-20).
4
Blood Biochemical Analysis Protocol
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After fresh blood collection, the serum was obtained by centrifugation at 3500 r/min for 5 min and stored at −20 °C. The stored samples were melted at room temperature and tested with the methods described below. Serum melatonin was measured with Agilent 6470 liquid chromatography-tandem MS (LC-MS-MS) (Agilent Technologies, Santa Clara, CA, USA). Non-esterified fatty acid (NEFA) was measured by NEFA kit (Randox Laboratories Ltd, Crumlin, Co. Antrim, UK), following the manufacturer’s instructions, the growth hormone was measured with a commercial ELISA (MBS703041; MyBiosource, San Diego CA, USA). Plasma was analyzed for glucose (catalog no. 439–90901; Wako Chemicals USA, Richmond, VA, USA) and prolactin by enzyme-linked immunosorbent assay (Bovine Prolactin ELISA; MBS2022462; MyBioSource.com). Progesterone was measured by radioimmune assay (RIA; KIP1458; Diasource, Nivelles, Belgium), and insulin was determined by a 125I-insulin RIA CT kit (CIS Bio International Ltd, Gif-Sur-Yvette, France). All measurements followed the manufacturer’s instructions.
5
Triglyceride Reduction in Acute Pancreatitis
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The primary outcome measure was the reduction in triglycerides within 24 h after admission, expressed as a percentage of the baseline value. Secondary outcomes were days needed to lower triglycerides below 10 mmol/L, highest CRP level during treatment and percentage of patients with a severe course of pancreatitis. Mortality was also recorded, as were the side effects of treatment (mainly hypocalcemia in the PE group and hypoglycemia in the insulin group). In a subgroup of patients, free fatty acids (FFA) were measured from available residual samples (NEFA kit, Randox, Crumlin, United Kingdom).
6
Plasma Metabolite Quantification
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Venous blood samples (∼6 mL) were collected into EDTA tubes, stored on ice, and then centrifuged at 4°C and 1,000 g for 15 min. Aliquots of plasma were then stored at −70°C and later analyzed for glucose, lactate, nonesterified fatty acids (NEFA), and glycerol using commercially available kits (Glucose kit, Lactate kit, NEFA kit, Glycerol kit; Randox, London, UK) using an automated photometric clinical chemistry analyzer RX Daytona+ (Randox, London, UK). Plasma galactose concentration was determined using a colorimetric assay (Galactose Assay Kit, Sigma Aldrich, St. Louis, MO) and insulin using an enzyme-linked immunosorbent assay (Invitrogen, Life Technologies, California).
7
Porcine Pancreatic Lipase Activity Assay
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An assay of PL activity was performed by standard method, described by Moreno et al. (2003) (link), with some modification. Briefly, different concentrations (250, 500, 1000, and 2000 mg/mL) of EEMUS, EEMUL and EESLC added separately to test tubes contain Tris-HCl buffer and incubated for 3 min at 37 °C and then 0.5 mL aliquot of porcine PL (250 mg/mL, type II, Sigma Chemical Co.) was added to each test tube to initiate the reactions. After 30 min of incubation, all the test tube was immersed in boiling water for 2 min to stop the reaction and then cooled. The free fatty acid concentration was determined according to manufacture protocol. using commercial kit (NEFA kit, Randox, China).
8
Plasma Metabolite and Isotope Analysis
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Plasma was analyzed for glucose, lactate, nonesterified fatty acids (NEFA), and glycerol with commercially available kits (Glucose kit, Lactate kit, NEFA kit, Glycerol kit; Randox, London, UK) using an automated photometric clinical chemistry analyzer RX Daytona+ (Randox, London, UK). Plasma galactose concentration was determined using a colorimetric assay (Galactose Assay Kit, Sigma Aldrich, St Louis. MO) and insulin using an enzyme-linked immunosorbent assay (Ultrasensitive Insulin, Mercodia, Uppsala Sweden). Breath samples were analyzed for 13C:12C ratio by gas chromatography isotope ratio mass spectrometry (IRMS), Hydra 20-20, Europa Scientific, Crewe, UK).
9
Blood Lipid and Metabolic Biomarker Analysis
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Serum concentrations of free fatty acids (FFAs), total cholesterol (TC), low density lipoprotein (LDL)-c, high density lipoprotein (HDL)-c and triglycerides (TGs) were analyzed using commercial kits (Randox Laboratories Ltd., Crumlin, UK) in accordance with the manufacturer's protocol, and a Hitachi 8060 automatic biochemical analyzer (Hitachi Co., Ltd, Tokyo, Japan). TC and TG levels were determined by CHOD-PAP and GPO-PAP colorimetric end-point assays, respectively, using Randox Total Cholesterol (cat. no., CH200) and Triglycerides (cat. no., TR1697) kits. FFAs were measured by the non-esterified fatty acids (NEFA) colorimetric method using a Randox NEFA kit (cat. no., FA115). HDL-c and LDL-c were quantified by a direct clearance method using Randox Direct HDL-c (cat. no., CH2652) and Direct LDL-c kits (cat. no., CH2656). Blood glucose was measured with a OneTouch® Ultra™ Blood Glucose Monitoring System. Serum insulin and adiponectin levels were determined using a rat/mouse insulin kit (Linco; EMD Millipore, Billerica, MA, USA) and adiponectin enzyme-linked immunosorbent assay kit (cat. no. MRP300; R&D Systems, Inc., Minneapolis, MN, USA) in accordance with the manufacturer's protocol.
10
Glucose and Lactate Metabolic Analysis
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An ABX Pentra 400 (Horiba ABX Diagnostics, Irvine, California) was used to measure media glucose and lactate concentrations, and to calculate glucose consumption and lactate efflux rates. Plasma nonesterified fatty acids (NEFA) were measured using a Randox NEFA kit (Randox Laboratories, Crumlin, County Antrim, United Kingdom).
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