Clinical trial NCT03982589: Telomere Length in Relation to Acute Stress Response in Critical Care Patients
Official title: Telomere Length in Relation to Acute Stress Response in Critical Care Patients
Study type: OBSERVATIONAL
Overall status: COMPLETED
Conditions: Acute Disease
Telomere Shortening
Lead sponsor: Rabin Medical Center
Sponsor class: OTHER
First posted: 2019-06-11
Enrollment: 40
Sex: ALL
Minimum age (years): 18
Maximum age (years): 75
Standard ages: ADULT
OLDER_ADULT
Accepts healthy volunteers: no
Observational model: COHORT
Time perspective: PROSPECTIVE
Sampling method: NON_PROBABILITY_SAMPLE
Study population: adults (male and females) admitted to the ICU

Brief summary: the investigators studied the impact of severe stress (in this case any event or illness leading to a necessity of critical care) on telomere length.

Detailed description: Telomeres length analysis were determined from 2 blood samples, the initial sample drawn in the first 72 hours after hospitalization and the second after not \< 5 or \> 14 days. For patients discharged before day 5, a repeat sample was drawn and analyzed on the discharge day. The blood sample processing was as follows: 5 ml of blood was collected and the red blood cells (RBC) were lysed using the RBC lysis solution (Biological Industries, Beit Haemek, Israel). Isolation of genomic DNA was performed by using the DNA isolation kit for mammalian blood (Roche, Mannheim, Germany). Briefly, DNA was isolated by the salting out procedure, washed and precipitated by isopropanol. The DNA was resuspended in polymerase chain reaction (PCR) grade water. The DNA concentration was measured by using the NanoDrop device (Thermo Fisher, USA).

DNA samples were analyzed for telomere length according to the method of Cawthon (2009) \[20\] with slight modifications. Each DNA sample was analyzed by two sets of primers detailed below, one for telomere length analysis and one for a reference gene analysis (human hemoglobin). The primers were diluted to 100µM in PCR grade water and then to 10µM. DNA samples were diluted to 2.5 ng/µl in PCR grade water. The primers sequences are shown below:

telc: TGTTAGGTATCCCTATCCCTATCCCTATCCCTATCCCTAACA telg: ACACTAAGGTTTGGGTTTGGGTTTGGGTTTGGGTTAGTGT hbgd:_GCCCGGCCCGCCGCGCCCGTCCCGCCGGAGGAGAAGTCTGCCGTT hbgu: GGCGGCGGGCGGCGCGGGCTGGGCGGCTTCATCCACGTTCACCTTG

Reaction PCR were processed as follows:

50°C for 2 min, 95°C for 5 min, a single cycle of 94°C for 15 sec, 49°C for 15 sec; 40 cycles of: 94°C for 15sec, 62°C for 10 sec and a final stage of: 74°C for 15 sec. All reactions were performed using the Step One device (ABI, USA).

Eligibility criteria: Inclusion Criteria:

* • Patients hospitalized up to 72 hours prior to admission to the ICU

  * Predicted ICU stay is at least 5 days

Exclusion Criteria:

* • Pregnancy and lactation

Primary outcomes:
measure: telomere length difference
timeFrame: 7 days
Secondary outcomes:
measure: correlation between mortality and telomere length
timeFrame: 6 month
measure: correlation between telomere length change and leukocytes count change
timeFrame: 7 days
Other outcomes:
Locations:
city: Petah Tikva
country: Israel
facility: Rabin medical center
Study officials:
References: