Blood glucose concentration was measured using HemoCue Glucose 201+. For blood glucose measurement, the patient forefinger was prepared as described for hemoglobin measurement above. A drop of blood was collected from a finger prick into a microcuvette and inserted into the HemoCue Glucose 201+ for blood glucose measurement. The measured blood glucose was later classified as normal (FBG < 100 mg/dL), prediabetes (FBG: 100–125 mg/dL), and diabetes (FBG > 126 mg/dL) [30 ].
Glucose 201
Manufactured by HemoCue
Sourced in Sweden, United Kingdom
The HemoCue Glucose 201+ is a point-of-care system designed for the quantitative determination of glucose in capillary, venous, and arterial whole blood. It provides rapid and accurate glucose measurements.
Automatically generated - may contain errors
Lab products found in correlation
Streptozotocin (stz), by Merck Group (3 mentions)
Hb 201, by HemoCue (2 mentions)
Humulin s, by Eli Lilly (2 mentions)
Nicotinamide, by Merck Group (2 mentions)
Nicotinamide na, by Merck Group (1 mentions)
Periflux 4001 master 4002 satellite system, by Perimed (1 mentions)
Cobas e411, by Roche (1 mentions)
K2edta tubes, by BD (1 mentions)
Sorvall st 16 centrifuge, by Thermo Fisher Scientific (1 mentions)
Cobas system, by Roche (1 mentions)
Bc 418ma, by Tanita (1 mentions)
Actrapid, by Novo Nordisk (1 mentions)
24 protocols using glucose 201
1
Blood Glucose Measurement Protocol
2
Measuring Glycemic Response to Cocoa
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GR was measured using the previously validated method of Wolever (2004) and redesigned by Coe et al. (2013) , and is in line with procedures recommended by the Food and Agriculture Organization/World Health Organization. A total of eight blood measures were taken using an automatic blood glucose analyser (Glucose 201+, Hemocue AB, Sweden).
Mean fasting BG was calculated using two 5µl finger-prick samples at -5 and 0 min, and measurements were collected at 15, 30, 45, 60, 90 and 120 minutes after the cocoa consumption. BG levels were compared to WHO guidelines to determine if any of the participants had fasting BG or postprandial BG values outside of the healthy range.
Mean fasting BG was calculated using two 5µl finger-prick samples at -5 and 0 min, and measurements were collected at 15, 30, 45, 60, 90 and 120 minutes after the cocoa consumption. BG levels were compared to WHO guidelines to determine if any of the participants had fasting BG or postprandial BG values outside of the healthy range.
3
Fasting Oral Glucose Tolerance Test
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For the collection of blood, a cannula is inserted into a forearm vein for sampling at time points 0, 15, 30, 45, 60, 90 and 120 min during the 75 g fsOGTT. Blood samples for downstream blood omics processing (as described above) are extracted and stored. A standard finger-stick fasting capillary blood glucose sample (with a HemoCue Glucose 201 or similar) is taken before the fsOGTT, and those with corrected fasting venous glucose >10 mmol/l (capillary glucose >11 mmol/l) are excluded from the study. To adjust for difference between plasma glucose and capillary blood glucose, a correction factor of ~1.11 is applied manually or automatically, as per most commercially available capillary blood glucose meters [27 (link)].
4
Whole Blood Hemoglobin and Glucose Measurement
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Whole blood hemoglobin was measured using a HemoCue® HB +201 portable analyzer, and glucose was measured using a HemoCue® Glucose 201 portable analyzer. The rest of the blood was placed on ice for up to 3 hr until it was centrifuged for 3 min at 12,000 g at Itasca Biological Station. Plasma and red blood cells were then stored separately in a −20°C freezer.
5
Standardized Glucose Measurement Protocol
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Methods for recording and definitions of educational level, current smoking, central obesity and blood pressure have been described elsewhere.9 (link)
A standard OGTT was performed in the morning after ≥10 hours fasting. Plasma glucose was analysed locally with a point-of-care technique (Glucose 201+, HemoCue, Ängelholm, Sweden). Values obtained with the HemoCue instrument were in 69% patients within 5%, in 91% patients within 10% and always within 14.3% of the ID Gas Chromatography - Mass Spectrometry (GC-MS) method.21 (link) The values were converted from whole venous blood to plasma applying the formula by Carstensen et al:22 (link) plasma glucose=0.558+0.119×whole blood glucose, as used by the Euro Heart Survey on Diabetes and the Heart.23 (link)
HbA1c was measured at the central laboratory (Disease Risk Unit, National Institute for Health and Welfare, Helsinki, Finland) with an immunoturbidimetric International Federation Clinical Chemists (IFCC) aligned method (Abbot Architect analyser; Abbott Laboratories, Abbott Park, Illinois, USA) in fasting venous whole blood sampled in an EDTA tube.
A standard OGTT was performed in the morning after ≥10 hours fasting. Plasma glucose was analysed locally with a point-of-care technique (Glucose 201+, HemoCue, Ängelholm, Sweden). Values obtained with the HemoCue instrument were in 69% patients within 5%, in 91% patients within 10% and always within 14.3% of the ID Gas Chromatography - Mass Spectrometry (GC-MS) method.21 (link) The values were converted from whole venous blood to plasma applying the formula by Carstensen et al:22 (link) plasma glucose=0.558+0.119×whole blood glucose, as used by the Euro Heart Survey on Diabetes and the Heart.23 (link)
HbA1c was measured at the central laboratory (Disease Risk Unit, National Institute for Health and Welfare, Helsinki, Finland) with an immunoturbidimetric International Federation Clinical Chemists (IFCC) aligned method (Abbot Architect analyser; Abbott Laboratories, Abbott Park, Illinois, USA) in fasting venous whole blood sampled in an EDTA tube.
6
Nestling Immune Response Assessment
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Feces and a small blood sample (<30 µL) were collected from nestlings when they were 10 days old. Hemoglobin was measured from whole blood using a HemoCue® HB +201 portable analyzer and glucose was measured using a HemoCue® Glucose 201 portable analyzer.
Blood samples were placed on ice until they were centrifuged for 3 minutes at 10000 rpm to separate the plasma from the red blood cells. Plasma and red blood cells were then stored separately in a -80°C freezer. Fecal samples were placed on ice until stored in a -80°C freezer.
Samples were then transported on dry ice to the University of Connecticut. Enzyme-linked immunosorbent assays (ELISA) and Tridelta PHASE haptoglobin assay (TP-801) were then performed to quantify P. sialia-binding antibody (IgY) and haptoglobin (acute phase protein) levels, respectively, in the plasma of parasitized nestling. IgY antibody levels from unsupplemented nestlings were first published in (Grab et al. 2019) ; see this paper for the detailed protocol for the IgY ELISA.
Blood samples were placed on ice until they were centrifuged for 3 minutes at 10000 rpm to separate the plasma from the red blood cells. Plasma and red blood cells were then stored separately in a -80°C freezer. Fecal samples were placed on ice until stored in a -80°C freezer.
Samples were then transported on dry ice to the University of Connecticut. Enzyme-linked immunosorbent assays (ELISA) and Tridelta PHASE haptoglobin assay (TP-801) were then performed to quantify P. sialia-binding antibody (IgY) and haptoglobin (acute phase protein) levels, respectively, in the plasma of parasitized nestling. IgY antibody levels from unsupplemented nestlings were first published in (Grab et al. 2019) ; see this paper for the detailed protocol for the IgY ELISA.
7
Inducing Type 2 Diabetes in Rats
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Type 2 diabetes was induced by a combination of Nicotinamide (NA, Sigma-Aldrich, Germany) and Streptozotocin (STZ, Sigma-Aldrich, Germany) at a dose of 200 mg/kg and 60 mg/kg of body weight, respectively, via two intraperitoneal injections over 15 min (15). To verify the diabetes induction, fasting blood sugar (FBS) was tested from the dorsal vein of the rats using a glucometer (HemoCue Glucose 201+, Sweden) three days after the injection. FBS 250 mg/dl was considered as diabetic animal models (16).
8
Microvascular Blood Flow Dynamics
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Local blood flow was quantified by LDF using the Periflux 4001 Master/4002 Satellite system (Perimed, Järfälla, Sweden) and expressed in arbitrary units (AU). The LDF probe was attached to the cleaned skin with an adhesive strip. Simultaneously, skin temperature was traced by a Peritemp device (Perimed, Järfälla, Sweden), blood pressure was non-invasively measured at the digital artery of the right hand middle finger (Finapres, Ohmeda, Englewood, USA). Blood glucose was measured by HemoCue Glucose201+ (HemoCue AB, Ängelholm, Sweden) after the acclimatization period, 20 min after glucose/water load (i.e., before the LDF was assessed), and at the end of the protocol.
9
Microvascular Reactivity After Oral Glucose Load
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Sixteen young healthy subjects, nonsmokers (21.4 ± 1.3 years old, 11 males, 5 females who were all in the follicular phase of their menstrual cycles) were enrolled in this study after informed written consent had been obtained. The study was approved by the local ethics committee (no. 0120-175/2017/6) and followed the recommendations of the Helsinki declaration and subsequent amendments for studies conducted in human subjects [31 (link)].
Before designing the study protocol for the LDF assessment, a preliminary experimental set enrolling 12 healthy participants was carried out to determine the time point corresponding to the highest blood glucose concentration after an OGL, which would then be chosen as the appropriate time to assess microvascular reactivity. Blood glucose was assessed spectrophotometrically by a portable glucometer (HemoCue Glucose201+, HemoCue AB, Ängelholm, Sweden) before (baseline) and after a standard OGL (75 g of glucose dissolved in 250 mL water); capillary blood samples were taken every ten minutes during a two hours period. We found that the highest glucose concentration was achieved after 35 min (8.9 ± 1.2 mmol/L, compared to baseline: 4.9 ± 0.4 mmol/L); in all subjects, glucose remained elevated for at least 60 min after OGL.
Before designing the study protocol for the LDF assessment, a preliminary experimental set enrolling 12 healthy participants was carried out to determine the time point corresponding to the highest blood glucose concentration after an OGL, which would then be chosen as the appropriate time to assess microvascular reactivity. Blood glucose was assessed spectrophotometrically by a portable glucometer (HemoCue Glucose201+, HemoCue AB, Ängelholm, Sweden) before (baseline) and after a standard OGL (75 g of glucose dissolved in 250 mL water); capillary blood samples were taken every ten minutes during a two hours period. We found that the highest glucose concentration was achieved after 35 min (8.9 ± 1.2 mmol/L, compared to baseline: 4.9 ± 0.4 mmol/L); in all subjects, glucose remained elevated for at least 60 min after OGL.
10
Blood glucose and insulin measurement protocol
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Blood glucose was measured by following the method described by Brouns et al. (31) . Participants were encouraged to warm their hand before a finger-prick to increase blood flow. Blood was expressed by massaging the finger gently from the base toward the tip to minimise plasma dilution. An aliquot of 5 µl blood sample was collected using an Abbott SF ® single-use lancing device (Owen Mumford). Capillary blood glucose was measured using calibrated HemoCue Glucose 201+ analysers (HemoCue ® Ltd). The capillary blood samples (300 µl) were collected into K 2 EDTA tubes (Becton, Dickinson and Company) for insulin analysis. Blood samples were stored on ice and the plasma was separated by centrifugation (10 min, 1000 g, 4°C) (Sorvall™ ST 16 Centrifuge; Thermo Fisher Scientific Inc.). The plasma was stored in microcentrifuge tubes at -80°C for future analysis. Plasma insulin was determined on the immunoassay analyzer Cobas e 411 (Roche Diagnostics GmbH) (32, 33) .
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