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16 protocols using bms213hs

1

Rheumatoid Arthritis Disease Activity Profiles

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The clinical data included age, sex, disease duration, 66/68-joint count, patient’s pain and global assessment (0~100 scale), physician’s global assessment (0~100 scale), Health Assessment Questionnaire Disability Index (HAQ-DI, 0~3), DAS28, SDAI and Clinical Disease Activity Index (CDAI). DAS28 scores were calculated by using the 66/68 joint count. Therefore, an involvement of over 28 joints was considered as the involvement of 28 joints. Patients were divided into two groups by disease activity at 24 weeks: high disease activity group (HDA at 24 weeks; DAS28>5.1) or remission, low and moderate disease activity group (non HDA at 24 weeks; DAS28≤5.1).
Laboratory data included rheumatoid factor (U/mL), ESR (mm/h), CRP (mg/dL), IL-6 (pg/mL), and soluble IL-6 receptor (sIL-6R) (ng/mL). The levels of IL‐6 were measured by high‐sensitivity Enzyme-Linked Immunosorbent Assay (ELISA) (BMS213HS; Bender MedSystems, Vienna, Austria) and sIL-6R by ELISA (BMS214; Bender MedSystems) according to the manufacturer’s instructions. Serum was collected every 4 weeks.
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2

Biomarker Quantification in Sleep Study

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Blood samples were taken at the end of the ten-minute supine rest after the sleep study. Samples were subjected to a routine laboratory procedure to have the serum separated. ET-1, CRP, IL-6, TNF-α, and VEGF were measured with Enzyme-Linked Immunosorbent Assay kits (ET-1, ADI-900-202A, Enzo Life Sciences Inc., New York, USA; CRP, 88-7502, eBioscience, Inc., California, USA; IL-6, BMS213HS, Bender MedSystems GmbH, Vienna, Austria; TNF-α, BMS223HS, Bender MedSystems GmbH, Vienna, Austria; VEGF, BMS277/2, Bender MedSystems GmbH, Vienna, Austria). The metabolites of NO were measured by the Griess reaction using prepared kits (ADI-917-020, Enzo Life Sciences Inc., New York, USA) before which samples were ultrafiltered through a 10,000 MWCO filter (Amicon® Ultra-15 10K, Merck Millipore Ltd., County Cork, Ireland) to remove protein as recommended.
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3

Biomarker Profiling of Inflammatory and Metabolic Factors

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The plasma concentration of TNFα, interleukin (IL)-6, IL-10, leptin, adiponectin, angiotensinogen, and irisin was measured using an enzyme-linked immunosorbent assay kit (eBioscience BMS223HS, BMS213HS, BMS215HS, Thermo Fisher Scientific, Waltham, MA, USA; Biovendor RD191001100, Brno, Czech Republic; Assaypro EA3501-1, St Charles, MO, USA; IBL International 27412, Hamburg, Germany; Biovendor RAG018R, Brno, Czech Republic). To determine malondialdehyde (MDA) levels, plasma samples were measured using the thiobarbituric acid-reactive substance (TBARS) method [17 (link)]. Briefly, we mixed samples with 20% acetic acid (pH 3.5), 0.8% thiobarbituric acid, and 8.1% sodium dodecyl sulfate and incubated the mixture at 95 °C for 1 h. After the mixture had cooled down to room temperature, trichloroacetic acid was added and detected spectrophotometrically at 532 nm.
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4

Quantification of Serum IL-6 Levels

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IL-6 was measured in serum samples of patients using a high-sensitivity IL-6 ELISA kit according to the manufacturer’s instructions (Thermo Fisher Scientific Cat# BMS213HS, RRID: AB_2575453, Thermo Fisher, Waltham, USA). Lower limit of detection (LLOD) according to the manufacturer 0.08 pg/ml.
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5

Quantifying IL-6 and IL-8 Levels

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Medium from cells treated with or without doxorubicin were collected. 50 μL samples were analyzed in IL-6 and IL-8 ELISA plates (Thermo Scientific, Cat # BMS213HS and KHC0081) and quantified according to standard curves.
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6

Quantification of Secreted Inflammatory Factors

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The quantification of the secreted levels of IL-1α (BMS243-2 Thermo Fisher Scientific, Waltham, MA, USA), PGE2 (EHPGE2, Thermo Fisher Scientific), IL-6 (BMS213HS, Thermo Fisher Scientific), MMP-1 (RAB0361, Sigma Aldrich), and TIMP-1(RAB0466, Sigma Aldrich) was performed using commercially available ELISA kits according to the manufacturer’s instructions.
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7

Quantifying Serum Inflammatory Markers

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IL6 and TNFα concentrations were measured in frozen serum using enzyme-linked immunosorbent assay (ELISA) systems. We used an aliquot for this purpose. In particular, TNFα and IL6 protein levels were estimated with a high sensitivity ELISA kit (#BMS223HS and #BMS213HS, Human IL-6 High Sensitivity ELISA, Thermo Fisher Scientific, Waltham, MA, United States), according to the manufacturers’ instructions. Serum TNFα and IL6 levels were expressed as pg/mL.
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8

Quantifying Inflammatory Cytokines in CM

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The collected conditioned media (CM) samples (Microglia preparation section) were tested for TNFα (Thermo Fisher Scientific, KHC3011), IL1β, IL 6 (Thermo Fisher Scientific, BMS213HS) and IFNγ were assessed using their respective ELISA kits, following the manufacturer's instructions. A Synergy 2 ELISA plate reader (BioTek, Winooski, VT, USA) was used to quantify ELISA signals. according to the manufacturer's instructions.
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9

Inflammatory Biomarkers in Response to Study Bread

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Inflammatory biomarkers, interleukin 8 (IL-8), interleukin 6 (IL-6) and lipopolysaccharide binding protein (LBP), were measured at baseline and after the treatment period with study breads. Serum IL-6 (BMS213HS, ThermoFisher Scientific, Waltham, MA USA), IL-8 (HS800, R&D Systems, Abingdon, UK) and LBP (HK315-01, LBP; Hycult Biotech, Uden, The Netherlands) levels were quantified using sensitive enzyme-linked immunosorbent assays (ELISA).
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10

Serum Cytokine Levels in Natalizumab Treatment

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Blood samples were taken on day 0 before natalizumab administration and 7 days afterwards (day 7). Blood was collected into serum tubes and transferred to the laboratory for storage within two hours. Serum samples were tested for concentrations of IL-6, IFN-γ, TNF-α. Ultrasensitive ELISA kits BMS213HS, BMS228HS and BMS223HS (Thermo Fisher) were used for quantitative detection of serum IL-6, IFN-γ and TNF-α respectively on both time points, following manufacturer's instructions. The detection limit that was used for statistical analysis was determined as OD blanco + SD x2, which is < 0.14 pg/ml for IFN-γ, < 0.16 pg/ml for IL-6 and < 1.25 pg/ml for TNF-α). Serum concentrations below the proposed detection limit were replaced by the arbitrary value determined as (OD blanco + SDx2)/2.
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