A guide for measurement of circulating metabolic hormones in rodents: Pitfalls during the pre-analytical phase

Bielohuby, Maximilian; Popp, Sarah; Bidlingmaier, Martin

doi:10.1016/j.molmet.2012.07.004

Cited by 44 publications

(40 citation statements)

References 58 publications

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“…The pre-analytical phase in laboratory medicine R e g a r d l e s s o f t h e m e t h o d o f m e a s u r e m e n t , t h e concentration an analyte in a biological sample depends on biological, analytical, and pre-analytical variability (11). Indeed, uses and lifestyle in the hours before sampling, sample drawing execution, handling, processing, and storing can cause unwanted changes in the concentration of certain types of biomolecules (e.g., degradation, release from cellular components or carrier molecules) (12,13).…”

Section: Pre-analytical Phase In the Assessment Of Myokines Involved mentioning

confidence: 99%

Measuring myokines with cardiovascular functions: pre-analytical variables affecting the analytical output

Lombardi¹,

Sansoni²,

Banfi³

2017

Ann. Transl. Med.

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Abstract:In the last few years, a growing number of molecules have been associated to an endocrine function of the skeletal muscle. Circulating myokine levels, in turn, have been associated with several pathophysiological conditions including the cardiovascular ones. However, data from different studies are often not completely comparable or even discordant. This would be due, at least in part, to the whole set of situations related to the preparation of the patient prior to blood sampling, blood sampling procedure, processing and/or store. This entire process constitutes the pre-analytical phase. The importance of the preanalytical phase is often not considered. However, in routine diagnostics, the 70% of the errors are in this phase. Moreover, errors during the pre-analytical phase are carried over in the analytical phase and affects the final output. In research, for example, when samples are collected over a long time and by different laboratories, a standardized procedure for sample collecting and the correct procedure for sample storage are acknowledged. In this review, we discuss the pre-analytical variables potentially affecting the measurement of myokines with cardiovascular functions.

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Section: Pre-analytical Phase In the Assessment Of Myokines Involved mentioning

confidence: 99%

Measuring myokines with cardiovascular functions: pre-analytical variables affecting the analytical output

Lombardi¹,

Sansoni²,

Banfi³

2017

Ann. Transl. Med.

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“…All blood samples were handled, processed, and stored as recommended previously (8). Fasting blood glucose and glucose during dynamic tests were measured using the glucose oxidase method (EcoSolo; Care Diagnostica, Voerde, Germany).…”

Section: Measurement Of Blood Glucose Lipids and Circulation Metabomentioning

confidence: 99%

Impaired glucose tolerance in rats fed low-carbohydrate, high-fat diets

Bielohuby

Sisley

Sandoval

et al. 2013

American Journal of Physiology-Endocrinology and Metabolism

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Tschöp MH, Seeley RJ, Bidlingmaier M. Impaired glucose tolerance in rats fed low-carbohydrate, high-fat diets. Am J Physiol Endocrinol Metab 305: E1059 -E1070, 2013. First published August 22, 2013; doi:10.1152/ajpendo.00208.2013.-Moderate low-carbohydrate/high-fat (LC-HF) diets are widely used to induce weight loss in overweight subjects, whereas extreme ketogenic LC-HF diets are used to treat neurological disorders like pediatric epilepsy. Usage of LC-HF diets for improvement of glucose metabolism is highly controversial; some studies suggest that LC-HF diets ameliorate glucose tolerance, whereas other investigations could not identify positive effects of these diets or reported impaired insulin sensitivity. Here, we investigate the effects of LC-HF diets on glucose and insulin metabolism in a well-characterized animal model. Male rats were fed isoenergetic or hypocaloric amounts of standard control diet, a high-protein "Atkinsstyle" LC-HF diet, or a low-protein, ketogenic, LC-HF diet. Both LC-HF diets induced lower fasting glucose and insulin levels associated with lower pancreatic ␤-cell volumes. However, dynamic challenge tests (oral and intraperitoneal glucose tolerance tests, insulintolerance tests, and hyperinsulinemic euglycemic clamps) revealed that LC-HF pair-fed rats exhibited impaired glucose tolerance and impaired hepatic and peripheral tissue insulin sensitivity, the latter potentially being mediated by elevated intramyocellular lipids. Adjusting visceral fat mass in LC-HF groups to that of controls by reducing the intake of LC-HF diets to 80% of the pair-fed groups did not prevent glucose intolerance. Taken together, these data show that lack of dietary carbohydrates leads to glucose intolerance and insulin resistance in rats despite causing a reduction in fasting glucose and insulin concentrations. Our results argue against a beneficial effect of LC-HF diets on glucose and insulin metabolism, at least under physiological conditions. Therefore, use of LC-HF diets for weight loss or other therapeutic purposes should be balanced against potentially harmful metabolic side effects. dietary intervention; macronutrients; Atkins-style and ketogenic diet; hyperinsulinemic euglycemic clamps; insulin resistance

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“…Therefore, serum has lower total protein content than plasma. Differing protein contents have been shown to result in differences in immunoassay results (Bielohuby et al, 2012). In addition, enzyme activity is lower in plasma than serum.…”

Section: Discussionmentioning

confidence: 99%

Influence of Collection and Storage Conditions on Adrenocorticotropic Hormone (ACTH) Measurements in Bottlenose Dolphins (Tursiops truncatus)

Lewis¹

2013

Aquat Mamm

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Identifying the effects of stressors on marine mammal health is critical for species conservation and collection. Adrenocorticotropic Hormone (ACTH) is a key indicator of peracute stress; however, handling methods may affect results. Thus, the aim of this study was to examine the effects of collection methods and storage temperature on the stability of bottlenose dolphin (Tursiops truncatus) ACTH measurements. Blood was collected from five managed-care bottlenose dolphins in four vacutainers ™ in the following sequence: EDTA, sodium heparin, serum separator, and 2nd EDTA. The specimens were centrifuged, processed immediately, aliquoted into plasma and serum, and divided into four equal volumes. These aliquots were immediately stored at either 4º or -80º C for 24 h prior to the ACTH assay. ACTH concentrations were compared between the collection tube types and the storage temperatures. There was a significant difference in ACTH concentration across collection tube types. ACTH concentrations were highest in ethylenediaminetetraacetic acid (EDTA) tubes and lowest in serum separator tubes at both temperatures. Twenty-four hour storage temperature did not have a statistically significant impact on mean ACTH concentration. There was no significant difference in ACTH concentration between the first and last drawn EDTA tubes. Bottlenose dolphin whole blood should be collected in EDTA for ACTH measurement. Samples may be refrigerated and shipped on cold packs if assayed within 24 h of collection or immediately frozen and shipped. When multiple samples are collected in various collection tubes at the same time, the order in which tubes are filled does not affect the ACTH measurement.

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A guide for measurement of circulating metabolic hormones in rodents: Pitfalls during the pre-analytical phase

Cited by 44 publications

References 58 publications

Measuring myokines with cardiovascular functions: pre-analytical variables affecting the analytical output

Measuring myokines with cardiovascular functions: pre-analytical variables affecting the analytical output

Impaired glucose tolerance in rats fed low-carbohydrate, high-fat diets

Influence of Collection and Storage Conditions on Adrenocorticotropic Hormone (ACTH) Measurements in Bottlenose Dolphins (Tursiops truncatus)

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