Clinical Trial Details
— Status: Completed
Administrative data
| NCT number |
NCT05752968 |
| Other study ID # |
111064 |
| Secondary ID |
|
| Status |
Completed |
| Phase |
|
| First received |
|
| Last updated |
|
| Start date |
May 3, 2023 |
| Est. completion date |
December 31, 2023 |
Study information
| Verified date |
February 2024 |
| Source |
Tungs' Taichung Metroharbour Hospital |
| Contact |
n/a |
| Is FDA regulated |
No |
| Health authority |
|
| Study type |
Observational
|
Clinical Trial Summary
Background
Peripheral arterial disease (PAD) and its relevant complications are more common in
hemodialysis (HD) patients. The potential association regarding chronic kidney disease
dysbiosis, inflammation and metabolic endotoxinemia in HD patients is unknown. A
cross-sectional study will be carried out the evaluate the possible association endotoxin
core antibody with asymptomatic PAD in a cohort of HD patients.
Methods
This cohort study enrolled 500 HD patients treated at a single center in Taichung city.
Fasting blood samples will be collected to determine biochemical data Endotoxin core antibody
levels and other related biomarkers. By the automatic oscillometric method, the
ankle-brachial index (ABI) was measured. Low ABI was defined as any value < 0.9.
Description:
Study Design and Patients This is a cross-sectional study involving hemodialysis patients
from the hemodialysis unit of Tungs' Taichung MetroHarbor Hospital, Taiwan. A total of 500
patients over 20 years of age who receive hemodialysis with ultrapure dialysate three times
weekly for at least 1 month are eligible for enrollment. We will exclude patients dialyzed
with a catheter; who had an acute infection requiring hospitalization; and who had underlying
malignancy or autoimmune diseases. In addition, patients who refuse to participate will be
excluded. Information on participant demographics and comorbidities will be obtained from
interviews and medical record reviews at the time of enrollment. Diabetes mellitus is defined
by self-reported history or use of antidiabetic medications. Hypertension was defined as
systolic blood pressure ≥140 mmHg, diastolic ≥90 mmHg, or use of antihypertensive agents.
Pre-exisiting CVD is defined as coronary artery disease, as documented by coronary
angiography or a history of myocardial infarction, class III or IV congestive heart failure,
or stroke. This study will comply with the Declaration of Helsinki and will apply for
approval from the Institutional Review Board of Tungs' Taichung MetroHarbor Hospital. All
participants will give their written informed consent.
Laboratory Measurements Predialysis blood samples are obtained on a mid-week day. Within 30
min after sampling, the remaining blood will be centrifuged at 3,000 g for 10 min,
immediately aliquoted and frozen at-70 °C until further analysis. The total serum cholesterol
is measured through the reaction of cholesterol esterase/cholesterol oxidase/ peroxidase,
using Hitachi 747 (Hitachi, Bohemia, NY, USA). HDL cholesterol is quantified after
precipitation with polyethylene glycol at room temperature. Serum glucose concentrations are
measured by the glucose oxidase method. Low-density lipoprotein cholesterol is calculated
using the Friedewald formula. Total serum triglycerides are measured through the reaction of
glycerol/ phosphate/oxidase and peroxidase. The serum levels of high-sensitivity C-reactive
protein (hsCRP) are measured using a Behring Nephelometer II (Dade Behring, Tokyo, Japan).
Serum levels of IL-6 and sCD14 are measured by a commercially available enzyme-linked
immunosorbent assay (Quantikine HS Immunoassay kit, R&D System, Minneapolis, MN, USA). The
LBP will be determined from serum samples and controls using standardized enzyme-linked
immunosorbent assay methods (HK315-02, HyCult Biotech Inc., Uden, The Netherlands), and serum
from 20 normal control subjects are used for interassay variation. Serum endotoxin core
antibody will be tested using a commercially available EndoCab IgG ELISA assay (Hycult
Biotech Inc., The Netherlands). In addition, serum LCN-2 (Lipocalin-2) and YKL-40
concentrations will be determined using enzyme linked immunosorbent assays
Endotoxin Measurement Endotoxin levels are measured using a Charles River Laboratories (CRL)
Kinetic Chromogenic assay, Endochrome-K. A more detailed description of the methodology used
has been described elsewhere [37]. Samples are thawed and diluted to 1 : 20 with a
bio-dispersing agent (CRL, BD100) and heat to 75°C for 15 minutes. A standard curve (0.005
EU/mL and 5 EU/mL) is prepared using control standard endotoxin supplied by CRL. If endotoxin
is detected, the samples are then reconstituted with an endotoxin-specific buffer (Charles
River Endosafe BG-120) to avoid any β-glucan-related false positive reaction. A positive
product control is included for each sample to eliminate the possibility of enhancement or
inhibition of endotoxin measurement. The absorbance at 405 nm is measured at 30 second
intervals for a total of 3600 seconds using a Biotek ELx808 plate reader. The assay reactions
are held at 37°C throughout the assay period. Charles River Endosafe, EndoScan V software is
used to generate a standard curve and analyze data using an onset time of 0.1 absorbance
unit.
