We collected resting saliva samples (i.e., without stimulation) in the morning (9–12 a.m.), after two hours at least of fasting, from all CF patients and controls. Resting saliva (about 1–3 mL) was collected in sterile plastic tubes in ice and then centrifuged for 30 min at 14,000 g to remove bacteria/cellular debris. In order to prevent in vitro auto-oxidation processes [28 (link)], the supernatants were spiked with butylated hydroxytoluene at the final concentration of 1 mg/mL and stored at −80 °C until analysis. Blood samples were also collected after an overnight fast from patients with CF. Serum was separated from blood cells and analyzed for serum total cholesterol and triglycerides determinations by an automated biochemistry analyzer (Architect ci16200 Integrated System, Abbott Diagnostics, Rome, Italy), as previously described [29 (link)].
Architect ci16200 integrated system
Manufactured by Abbott
Sourced in United States, Italy
The ARCHITECT ci16200 Integrated System is a fully automated clinical chemistry and immunoassay analyzer designed for high-volume laboratories. It offers a comprehensive menu of diagnostic tests and is capable of performing a wide range of analyses on various sample types. The system is equipped with advanced technology to ensure accurate and reliable results.
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Cobas e610, by Roche (1 mentions)
Glu reagent kit, by Roche (1 mentions)
Modular p, by Roche (1 mentions)
Model hem 752 fuzzy, by Omron (1 mentions)
Advia centaur xp, by Siemens (1 mentions)
20 protocols using architect ci16200 integrated system
1
Saliva and Serum Analysis in Cystic Fibrosis
2
Serum Vitamin Analysis in ETI Therapy
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Blood samples were collected from fasted patients before ETI treatment (baseline) and after 3 and 12 months of treatment. Serum samples were separated from blood cells and then analyzed for biochemical parameters by an automated biochemistry analyzer (Architect CI-16200 Integrated System, Abbott Diagnostics, Rome, Italy), as previously described [24 (link),25 (link)]. All procedures were performed under internal and external laboratory quality control assurance. Serum vitamins A and E were analyzed by an isocratic high-performance liquid chromatography (HPLC) method (Vitamin A/E by HPLC assay, Bio-Rad Laboratories, Segrate, Italy) with an HPLC-UV system (Agilent 1260 Infinity Quaternary LC coupled with BIO-RAD UV-1806 detector, Bio-Rad Laboratories, Segrate, Italy).
3
Serum Biomarker and Flow Cytometry Analysis
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Serum samples were separated from blood cells after the collection in tubes without anticoagulant and stored at − 80 °C until analysis. Serum IL-6, MPO, MMP3 and MMP9 were analyzed by Human Magnetic Luminex Assay on Biorad Bio-Plex 100 system (Labospace s.r.l., Milan, Italy). Serum IL-17A was measured using specific human ELISA Max™ Set Deluxe kit (BioLegend, Inc., San Diego, USA), in accordance with the manufacturer's instructions. The serum hs-CRP was determined by a commercial kit (Abbott Diagnostics, Rome, Italy) and an automated biochemistry analyzer (Architect ci 16200 Integrated System, Abbott Diagnostics, Rome, Italy).
For cytometric analysis, the whole blood samples were collected in tubes containing EDTA and then analyzed by Facs Canto II (Becton Dickinson Italia, Milan, Italy). Lymphocytes, neutrophils, and monocytes were firstly separated on the basis of forward scatter and sideward scatter characteristics. In addition, the cells were gated with CD45 and sideward scatter21 (link).
For cytometric analysis, the whole blood samples were collected in tubes containing EDTA and then analyzed by Facs Canto II (Becton Dickinson Italia, Milan, Italy). Lymphocytes, neutrophils, and monocytes were firstly separated on the basis of forward scatter and sideward scatter characteristics. In addition, the cells were gated with CD45 and sideward scatter21 (link).
4
CRP and Salivary Cytokine Analysis
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Serum CRP was measured by a commercial kit (Abbott Diagnostics, Rome, Italy) with an automated biochemistry analyzer (Architect ci 16,200 Integrated System, Abbott Diagnostics, Rome, Italy). Salivary cytokines were analyzed using human IL-6, IL-8, and TNFα ELISA Max™ Set Deluxe kits (BioLegend, Inc., San Diego, CA, USA), as previously described20 .
5
Anthropometric and Metabolic Profiling
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We recorded height, weight, and BMI. Anthropometric measurements were performed in triplicate according to Cameron [20 ]. Body weight, expressed in kilograms, was measured at fasting state in the morning with a mechanical balance (± 0.1 kg, SECA 700, Hamburg DE). BMI was calculated as body weight divided by height squared (kg/m2) with categories in accordance with the WHO guidelines. Blood samples from all participants were collected in the morning after an overnight fast at the time of sampling for routine purposes. Serum samples were separated and analyzed as follows: serum albumin was measured by colorimetric method (Albumin BCG assay; Abbott Diagnostics, Rome, Italy); glucose, total cholesterol, and triglyceride levels were analyzed using specific enzymatic assays; and HDL and LDL cholesterol levels were measured using Ultra HDL methods and Multigent Direct LDL, respectively (Abbott Diagnostics, Rome, Italy). All assays were performed using an automated biochemistry analyzer (Architect ci16200 Integrated System; Abbott Diagnostics, Rome, Italy).
6
Salivary Biomarker Analysis Methods
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Salivary concentrations of chloride and potassium were measured using Integrated Chip Technology (Abbott Diagnostics, Rome, Italy). All other assays were performed using reagent kits provided by Abbott Diagnostics with an automated biochemistry analyzer (Architect ci16200 Integrated System, Abbott Diagnostics, Rome, Italy). In particular, salivary calcium and phosphate were measured by colorimetric methods. Salivary lactate dehydrogenase (LDH) was analyzed by the specific enzymatic assay. Total proteins were measured by the urine/CSF protein method.
7
Biochemical Markers in Fasting Serum
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Blood samples (n = 214) were collected from the participants after an overnight fast, and serum samples were separated. Serum albumin, insulin, C-reactive protein, glucose, total cholesterol, HDL and LDL cholesterol, and triglycerides were measured as previously described [6 (link)]; all assays were performed with an automated biochemistry analyser (Architect ci16200 Integrated System, Abbott Diagnostics, Rome, Italy). The HOMA index was calculated as fasting glucose × fasting insulin/22.5 [23 (link)]. Concentrations of total adiponectin in serum were measured in triplicate by an enzyme-linked immunosorbent assay (ELISA) as previously described [25 (link)]. A Sandwich-ELISA assay was used to measure leptin concentration in serum samples (Elabscience, Houston, Texas, USA).
8
Serum Metabolic Profile in Cystic Fibrosis
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Plasma and serum samples were collected from fasted patients. For homo-deltaF508 patients we collected the samples before and after lumacaftor/ivacaftor treatment. The period of treatment (from 2017 to 2019) ranged from 9 to 29 months (mean (SD): 15 (6) months). Serum samples were separated from blood cells and then analyzed for biochemical parameters by an automated biochemistry analyzer (Architect ci 16,200 Integrated System, Abbott Diagnostics, Rome, Italy) as previously described [ 17 Elce A. Nigro
9
Comprehensive Metabolic Profiling of Adults
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All study visits took place in the morning; at these visits, all subjects provided information on their medical history and lifestyle (such as dietary habits, smoking status, alcohol consumption, and exercise) through a questionnaire administered by trained interviewers. Height and weight were measured with an electronic scale and a wall-mounted stadiometer. The body mass index (BMI) was calculated using weight (kg) divided by the square of height (m2). Systolic blood pressure (SBP) and diastolic blood pressure (DBP) were measured twice in the right arm in a sitting position after a 15-minute rest, and the two replicate measurements were averaged. Blood samples were collected from the subjects after an overnight fast. Laboratory measurements of alanine aminotransferase (ALT), aspartate aminotransferase (AST), fasting plasma glucose (FPG), triglycerides (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), hepatitis B surface antigen and anti-hepatitis C virus antibodies, were performed using an ARCHITECT ci16200 Integrated System (Abbott, Illinois, USA).
10
Chronic Kidney Disease and Metabolic Syndrome
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eGFR < 60 mL/min/1.73 m2 was considered as CKD based on the Kidney Disease Outcomes Quality Initiative provided by the USA National Kidney Foundation18 . Serum creatinine levels were assayed with the picric acid method using the automatic analyzer (ARCHITECT ci16200 Integrated System; Abbott Laboratories, Abbott Park, Illinois, USA) at the standardized laboratory of Ruijin Hospital in Shanghai, China.
According to the Chinese Diabetes Society criteria in 200419 , MetS was diagnosed when at least three of the following disorders occurred: (i) overweight and/or obesity – BMI ≥ 25.0 kg/m2; (ii) hyperglycemia – fasting plasma glucose ≥6.1 mmol/L and/or 2‐h plasma glucose ≥7.8 mmol/L or diagnosed as type 2 diabetes mellitus before or received medicine; (iii) hypertension – systolic/diastolic blood pressure ≥140/90 mmHg or diagnosed hypertensive before and received medicine; and (iv) dyslipidemia – triglyceride level ≥1.7 mmol/L and/or high‐density lipoprotein cholesterol level <0.9 mmol/L (men) or <1.0 mmol/L (women).
According to the Chinese Diabetes Society criteria in 2004
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