Background
Materials & methods
Study design, subjects & ethics information
Definition of sepsis
Definition of AKI
Determination of vancomycin and gentamicin serum concentrations
Data and sample collection
Determination of NGAL, IL-6, PCT, and Screa
Determination and isolation of miRNAs
miRNA isolation
RTqPCR screening
Sample number/days 1, 4, or 7 and treatment | Total RNA (ng/μL) | Initial concentration to RT (ng/μL) | Dilution factor (×) |
---|---|---|---|
1/1 gentamicin | 173.3 | 5 | 35 |
1/4 | 235.9 | 10 | 24 |
1/7 | 459.3 | 10 | 46 |
2/1 gentamicin | 78.4 | 5 | 16 |
2/4 | 72.3 | 5 | 14 |
2/7 | 72.4 | 5 | 14 |
3/1 vancomycin | 166.3 | 5 | 33 |
3/4 | 167.6 | 5 | 34 |
3/7 | 230.3 | 10 | 23 |
4/1 vancomycin | 170.3 | 5 | 34 |
4/4 | 134.1 | 5 | 27 |
4/7 | 175 | 5 | 35 |
5/1 other ATB | 203.2 | 10 | 20 |
5/4 | 1084.5 | 10 | 108 |
5/7 | 1459.5 | 10 | 146 |
6/1 other ATB | 135.1 | 5 | 27 |
6/4 | 152.6 | 5 | 31 |
6/7 | 109.8 | 5 | 22 |
7/1 gentamicin | 146.5 | 5 | 29 |
7/4 | 263.3 | 10 | 26 |
7/7 | 435.8 | 10 | 44 |
8/1 other ATB | 192.8 | 10 | 19 |
8/4 | 222.8 | 10 | 22 |
8/7 | 360 | 10 | 36 |
hsa-miRNA | 133a-3p | 15a-5p | 16-5p | 4270 | 218–1-3p | 320b | 203a-3p | 138-3p | 489-3p | 301a-5p | 423-5p | 4321 | 192-5p |
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
¢Ct | 35.2 | 18.8 | 34.6 | 30.8 | 33.6 | 19.1 | 32.4 | 36.2 | 36.1 | 29.7 | 18.3 | 39.1 | 19.9 |
max Cq | 35.8 | 22.3 | 37.1 | 31.5 | 35.7 | 20.8 | 34.1 | 38.9 | 37.6 | 31.3 | 19.6 | 40.4 | 22.8 |
min Cq | 33.8 | 14.6 | 31.2 | 29.1 | 31.9 | 17.0 | 30.4 | 33.6 | 33.6 | 27.9 | 17.1 | 36.7 | 18.4 |
Gene | Expression viability value |
---|---|
hsa-miR-155-5p | 0.073 |
hsa-miR-320a | 0.077 |
hsa-miR-486-5p | 0.088 |
cel-miR-39-3p | 0.081 |
hsa-miR-223-5p | 0.099 |
hsa-miR-142-3p | 0.111 |
miREA
Statistical analysis
Results
Clinical, demographic and biochemical characteristics of patients
Vancomycin group | Gentamicin group | oATB group | P | |
---|---|---|---|---|
N (%) | 7 (15) | 20 (43) | 19 (41) | |
Age (years) | 65 (47–76) | 59 (47–72) | 65 (63–68) | 0.785 |
Male n (%) | 4 (57) | 13 (65) | 13 (68) | 0.866 |
BMI (kg/m2) | 26.2 (23.3–27.8) | 28.4 (24.0–32.5) | 26.3 (24.7–30.9) | 0.620 |
SOFA score (%) | 10 (7–14) + 1 QuickSOFA | 8 (5–12) | 7 (3–10) | 0.406 |
AKI stage at baseline/end of study, all n (%) | ||||
1 | 1 (14) / 1 (14) | 5 (25) / 3 (15) | 4 (21) / 3 (16) | 0.836/0.995 |
2 | N | 5 (25) / 1 (20) | 6 (32) / 0 | 0.243/0.515 |
3 | 2 (29) / 2 (29) | 2 (10) /5 (5) | 6 (32) / 9 (47) | 0.236/0.318 |
No AKI | 4 (57) / 4 (57) | 8 (40) / 11 (55) | 3 (15) / 7 (37) | 0.088/0.455 |
RRT initiation, n (%) | 2 (2) | 5 (25) | 7 (37) | 0.719 |
Use of vasopressors, n (%) | 6 (86) | 15 (75) | 11 (58) | 0.307 |
Type of vasopressor, n (%) | ||||
Norepinephrine | 6 (100) | 14 (93) | 8 (73) | 0.172 |
Dobutamine | N | N | 1 (9) | 0.484 |
Dopamine | N | N | 1 (9) | 0.484 |
Norepinephrine + Dobutamine | N | 1 (7) | 1 (9) | 0.416 |
Dose of vasopressors (μg/kg/min) | ||||
Norepinephrine | 0.18 (0.14–0.23) | 0.21 (0.17–0.29) | 0.32 (0.22–0.45) | 0.104 |
Dobutamine | N | N | 1.79 (1.19–2.38) | 0.333 |
Dopamine | N | N | 24.41 (19.41–29.41) | 0.333 |
Norepinephrine / Dobutamine | N / N | 0.14 (0.001–0.28) / 1.04 (0.69–1.39) | 0.24 (0.002–0.48) / 11.63 (0.16–23.10) | 0.467/0.600 |
Duration of vasopressors therapy (days) | 10 (6–12) | 6 (2–9) | 6 (3–13) | 0.256 |
ICU hospitalized patients, n (%) | 6 (86) | 18 (90) | 15 (79) | 0.629 |
ICU length of stay (days) | 18 (11–38) | 13 (9–23) | 12 (10–22) | 0.585 |
Intermediate care hospitalized patients, n (%) | 1 (14) | 2 (10) | 4 (21) | 0.629 |
Intermediate care length of stay (days) | 12 | 9 (8–10) | 16 (10–24) | 0.438 |
Total length of hospitalization (days) | 31 (12–52) | 25 (15–35) | 17 (12–24) | 0.162 |
Diagnoses, all n (%) | ||||
Urogenital infection | 2 (29) | 4 (20) | 5 (26) | 0.856 |
Pneumonia | ||||
- Nosocomial | 1 (14) | 2 (10) | 2 (11) | 0.950 |
- Community acquired | N | N | 2 (11) | 0.226 |
Sepsis in severe trauma | 1 (14) | 4 (20) | N | 0.127 |
Peritonitis | 1 (14) | 6 (30) | 1 (5) | 0.122 |
Mediastinitis | 1 (14) | 1 (5) | N | 0.280 |
Liver abscess | 1 (14) | N | N | 0.058 |
Acute cholecystitis | N | N | 1 (5) | 0.484 |
Acute pancreatitis | N | N | 1 (5) | 0.484 |
Infectious gastroenteritis | N | N | 1 (5) | 0.484 |
Abscess or phlegmon | N | 2 (10) | 3 (16) | 0.511 |
Erysipelas | N | N | 1 (5) | 0.484 |
Sepsis of unknown origin | N | 1 (5) | 2 (11) | 0.587 |
Clinical outcome survived/died, n (%) | 6 (86) / 1 (14) | 18 (90) / 2 (10) | 15 (79) / 4 (21) | 0.629 |
Parameter | Day | Vancomycin | IQR | Gentamicin | IQR | Other ATBs | IQR | P |
---|---|---|---|---|---|---|---|---|
NGAL (μg/L) | 1 | 240 | 120–1150 | 290 | 136–937 | 468 | 210–775 | 0.558 |
NGAL (μg/L) | 4 | 170 | 100–520 | 217 | 105–351 | 438 | 292–488 | 0.105 |
NGAL (μg/L) | 7 | 130 | 75–430 | 157 | 125–476 | 358 | 200–583 | 0.172 |
Screa (μmol/L) | 1 | 71 | 59–165 | 116 | 87–166 | 156 | 131–239 | 0.063 |
Screa (μmol/L) | 4 | 59 | 45–139 | 85 | 68–98 | 129 | 75–226 | 0.064 |
Screa (μmol/L) | 7 | 62 | 42–225 | 78 | 57–108 | 101 | 66–160 | 0.378 |
IL-6 (ng/L) | 1 | 51 | 31–109 | 310 | 61–5500 | 176 | 73–1073 | 0.107 |
IL-6 (ng/L) | 4 | 34 | 13–109 | 47 | 19–185 | 55 | 25–108 | 0.598 |
IL-6 (ng/L) | 7 | 23 | 11–92 | 27 | 12–152 | 66 | 20–289 | 0.551 |
PCT (μg/L) | 1 | 0.5 | 0.2–9.8 | 7.9 | 0.7–22.7 | 2.5 | 1.5–5.9 | 0.162 |
PCT (μg/L) | 4 | 0.3 | 0.1–2.7 | 1.5 | 0.3–1.8 | 1.5 | 0.5–5.1 | 0.297 |
PCT (μg/L) | 7 | 0.2 | 0.0–3.0 | 0.7 | 0.1–1.8 | 0.7 | 0.2–1.3 | 0.530 |
Selected miRNAs expression
miRNAs expression and relationship with biochemical parameters
Group | Parameter | Spearman r | hsa-miR-155-5p | hsa- miR-192-5p | hsa-miR-423-5p |
---|---|---|---|---|---|
Day 4 | |||||
Vancomycin | Screa | rs | 0.786 | – | – |
P | 0.048 | – | – | ||
NGAL | rs | 0.821 | – | – | |
P | 0.034 | – | – | ||
oATB | IL-6 | rs | – | 0.642 | – |
P | – | 0.003 | – | ||
all patients | NGAL | rs | 0.351 | 0.339 | 0.419 |
P | 0.017 | 0.021 | 0.004 | ||
IL-6 | rs | 0.328 | – | – | |
P | 0.026 | – | – | ||
Day 7 | |||||
Vancomycin | Screa | rs | – | 0.821 | – |
P | – | 0.034 | – | ||
NGAL | rs | – | 0.929 | – | |
P | – | 0.007 | – | ||
PCT | rs | – | 0.847 | – | |
P | – | 0.025 | – | ||
oATB | PCT | rs | – | 0.558 | 0.643 |
P | – | 0.016 | 0.004 | ||
IL-6 | rs | – | 0.598 | 0.719 | |
P | – | 0.009 | < 0.001 | ||
all patients | PCT | rs | 0.413 | 0.413 | 0.550 |
P | 0.005 | 0.005 | < 0.001 | ||
IL-6 | rs | 0.357 | – | 0.491 | |
P | 0.017 | – | < 0.001 | ||
NGAL | rs | 0.523 | – | 0,421 | |
P | < 0.001 | – | 0.004 | ||
Screa | rs | 0.419 | – | 0.313 | |
P | 0.004 | – | 0.036 |
Group | miRNA | No AKI | AKI | |||
---|---|---|---|---|---|---|
Median | IQR | Median | IQR | P | ||
Vancomycin | hsa-miR-15a-5p (amol/L) | 132.7 | 105.9–228.2 | 228.4 | 144.9–368.3 | 0.229 |
hsa-miR-155-5p (amol/L) | 4.3 | 3.7–4.7 | 7.4 | 3.5–26.6 | 0.400 | |
hsa-miR-192-5p (amol/L) | 21.4 | 10.4–68.6 | 96.8 | 69.9–101.8 | 0.114 | |
hsa-miR-423-5p (amol/L) | 640.4 | 407.5–895.0 | 1004.0 | 768.5–3313.0 | 0.229 | |
Gentamicin | hsa-miR-15a-5p (amol/L) | 274.7 | 125.5–467.0 | 109.6 | 75.1–182.6 | 0.013 |
hsa-miR-155-5p (amol/L) | 4.5 | 3.0–5.8 | 5.5 | 4.2–6.5 | 0.182 | |
hsa-miR-192-5p (amol/L) | 33.4 | 25.1–62.7 | 30.4 | 10.9–63.9 | 0.824 | |
hsa-miR-423-5p (amol/L) | 1024.0 | 517.2–1156.0 | 1016.0 | 634.9–1651 | 0.552 | |
oATB | hsa-miR-15a-5p (amol/L) | 171.2 | 137.4–292.0 | 277.8 | 222.0–432.6 | 0.299 |
hsa-miR-155-5p (amol/L) | 4.9 | 3.7–6.9 | 6.9 | 4.4–10.0 | 0.270 | |
hsa-miR-192-5p (amol/L) | 32.8 | 26.4–85.3 | 81.9 | 34.6–125.6 | 0.227 | |
hsa-miR-423-5p (amol/L) | 668.7 | 513.3–973.1 | 812.0 | 493.3–2358.0 | 0.592 |