Blood samples were obtained at each visit and shipped to a central laboratory for HbA1c assay (Tosoh A1c 2.2 Plus Glycohemoglobin Analyzer™, Tosoh Medics, South San Francisco, CA), reference range, 4%–6%. Simultaneous samples were processed with the DCA-2000 (Siemens Healthcare Diagnostics, Deerfield, IL) on site. These results were used to impute replacement values if samples were lost or damaged (1.2% of values).16 Data on demographic and disease-related characteristics were obtained from the electronic medical record and from parent report. Family income was reported by parents and categorized as <$50,000 (low), $50,000 to <$100,000 (middle), and ≥$100,000 (high).
Dca 2000
Manufactured by Siemens
Sourced in United States, United Kingdom
The DCA 2000 is a laboratory equipment product from Siemens. It is designed to perform specific functions within a laboratory setting. Without making any interpretations or extrapolations, the core function of the DCA 2000 is to provide accurate and reliable measurements for laboratory analysis.
Automatically generated - may contain errors
10 protocols using dca 2000
1
HbA1c Assay and Demographic Data Collection
2
Glycemic Control and Hemostasis Evaluation
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Blood samples were obtained between 8 and 10 AM by venipuncture without a tourniquet after overnight fasting. Platelet count was assessed using anticoagulated blood collected with a 10% ethylenediamine tetraacetic acid additive using a blood cell counter (Sysmex KX 21, Kobe, Japan). Platelet-poor plasma was collected with 3.2% (0.109 M) sodium citrate at a 1:9 ratio and centrifuged at 3,000 rpm for 15 minutes. PT, aPTT, and Fg tests were immediately carried out using an ACL 300 coagulation analyzer (Diamond Diagnostics, Holliston, MA, USA). The PAI-1 and vWF:Ag plasma levels were measured by enzyme-linked immunosorbent assay (Asserachrom, Diagnostica Stago, Asnières-sur-Seine, France). Glycemic control was evaluated using hemoglobin A1c (HbA1c; DCA 2000, Siemens, USA), fasting blood glucose (enzymatic method, Wiener Lab, Rosario, Argentina), and capillary blood glucose levels. According to the recommendation by the ADA, HbA1c < 8% value was considered to represent good glycemic control (GGC), while HbA1c ≥ 8% indicated poor glycemic control (PGC) [13 (link)].
3
Analyzing Metabolic Markers in Cohort
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HbA1c was analysed locally at each centre, using DCCT aligned methods (HbA1c Variant analyser [Bio-Rad Laboratories, Hercules, CA, USA], Adams Arkray [Kyoto, Japan], Vantage analyser [Siemens Diagnostics, Camberley, UK] or DCA 2000, [Siemens Diagnostics, Tarrytown, NY, USA]). Lipid profile measurements (cholesterol, HDL-cholesterol, LDL-cholesterol, triacylglycerol) were measured using routine laboratory methods.
The standard deviation score (SDS) was calculated for height, weight and BMI according to the least-mean squares method [17 (link)]. Blood pressure was measured using an Omran M6 blood pressure monitor (Hoofddorp, the Netherlands) and/or Dinamap monitor (Tampa, FL, USA) using an appropriate sized cuff. Age- and sex-related percentiles and SDS for systolic blood pressure (SBP) and diastolic blood pressure (DBP) were calculated according to published standards [18 (link)].
This study was approved by the Human Research Ethics Committees of each participating centre. Informed consent was obtained from participants and their families.
The standard deviation score (SDS) was calculated for height, weight and BMI according to the least-mean squares method [17 (link)]. Blood pressure was measured using an Omran M6 blood pressure monitor (Hoofddorp, the Netherlands) and/or Dinamap monitor (Tampa, FL, USA) using an appropriate sized cuff. Age- and sex-related percentiles and SDS for systolic blood pressure (SBP) and diastolic blood pressure (DBP) were calculated according to published standards [18 (link)].
This study was approved by the Human Research Ethics Committees of each participating centre. Informed consent was obtained from participants and their families.
4
Glycemic Control Measurement Protocol
5
Longitudinal Metabolic and Immunological Profiling
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Physical examination was performed and metabolic and immunological samples obtained at baseline; weeks 1, 2, 4, 6, 8 and 10; and months 3, 6, 9, 12, 18, and 24 from the time of enrollment. The following were performed at each visit. Glycated hemoglobin (HbA1c) was measured via DCA2000 or DCA Vantage Analyzer (Siemens Healthcare Diagnostics, Malvern, PA). MMTT (Boost, 6 mL/kg, maximum 360 mL) was administered, serum C-peptide was measured at the Northwest Lipid Research Laboratories, and 2-h C-peptide area under the curve (AUC) was calculated. Autoantibodies against glutamic acid decarboxylase, insulinoma-associated protein 2, insulin, and zinc transporter 8 were measured via radioimmunoassay at the Barbara Davis Center (13 (link)–15 (link)). Flow cytometric analysis of peripheral blood immune cell subsets was performed as described below. Adverse events (AEs) were recorded in accordance with Common Terminology Criteria for Adverse Events classifications (version 4.0; http://evs.nci.nih.gov/ftp1/CTCAE/CTCAE_4.03_2010-06-14_QuickReference_5x7.pdf ).
6
Standardized Risk Assessment for T2DM
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According to a standard operational policy (SOP), all recruited individuals attended a standardised health assessment appointment with an occupational health nurse which lasted 30-40 min. During the session, demographic (date of birth, gender and postcode of residence) and anthropometric (body mass, height and waist circumference) data were collected; systolic and diastolic blood pressure, smoking status, lifestyle choices (physical activity and dietary information) and family and medical histories were all recorded. In addition, a capillary blood sample was obtained and analysed immediately for HbA 1c (DCA 2000; Siemens Healthcare Diagnostics Ltd, Frimley, UK). The full details of the risk assessment have been published in detail elsewhere. 12 The risk of developing T2DM was predicted by the QDiabetes, 13 Leicester Risk Assessment, 14 FINDRISC 15 and Cambridge Risk Score 16 models.
7
Measuring HbA1c and C-peptide in diabetes
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HbA 1c levels were measured by the DCA 2000 (Siemens Health Care) point-ofservice method at all of the centers. Random C-peptide concentration was obtained #28 days of diagnosis and was available in a subset of the study population (n = 631), some of whom had participated in an ancillary study of the PDC Registry (16) . That study measured Cpeptide by a two-site immunoenzymatic assay using a Tosoh 2000 auto-analyzer (TOSOH, Biosciences, South San Francisco, CA) at the Northwest Lipid Research Laboratory (University of Washington, Seattle, WA). Otherwise, C-peptide levels were obtained from the local site databases.
8
Glycemic Control and Variability in Diabetes
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Statistical analyses were performed using SAS software version 9.2. The first 12 h of CGM data were removed from the analyses since the accuracy of CGM data is often poor initially. If more than 20% of the data were missing on any given day, the data for that day were also excluded. Measures of glycemic control included HbA1c (DCA2000, Siemens, Bayer Corporation, Elkhart, IN), the overall mean of glucose values, percentage of values above 140 or 200 mg/dL, as well as area under the curve (AUC) of glucose calculated by the trapezoidal rule. Primary variables to characterize glycemic variability included glucose range, the overall SD, and the coefficient of variation (CV). A two-tailed P value with an α-level for significance was set at 0.05. We used Wilcoxon–Mann–Whitney tests to compare CGM statistics between groups. We calculated the individual means first and then compared the mean of the individual means between the groups. Additionally, we calculated the AUC of the receiver operating characteristic curve for association with the development of diabetes among Ab+ subjects.
9
Comprehensive Health Assessments in Occupational Settings
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During normal working hours, individuals attended the occupational health department for an appointment lasting 60 min. During the visit, demographic (date of birth, gender, postcode of residence) and anthropometric (body mass, height, waist circumference) data were collected. Systolic and diastolic blood pressure, heart rate and rhythm (to screen for cardiac arrhythmias), smoking status, dietary habits (fruit and vegetable consumption), family and medical histories were recorded and blood samples obtained. Blood samples were collected via capillary puncture and analysed for total, high-density lipoprotein (HDL) and low-density lipoprotein (LDL) cholesterol; triglycerides; blood glucose (Cholestech LDX; Alere Inc., Orlando, FL, USA); and HbA 1c (DCA 2000; Siemens Healthcare Diagnostics Ltd, Frimley, UK). In addition, current physical activity levels were assessed by the General Practice Physical Activity Questionnaire (GPPAQ). 14
10
Urinary Albumin Quantification
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Urinary albumin levels were measured by DCA 2000 (Siemens).
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