Background
Material and methods
Ethics statement
The mother-child cohort in Crete, Rhea study
Definition of the outcomes
Metabolic syndrome variables
1H NMR spectroscopic analysis of urine
Sample handling and preparation
1H NMR experiments and data processing
Statistical analysis
Results
Descriptive statistics of the study population
SPB (n = 88) | IPB (n = 26) | FGR (n = 36) | SGA (n = 19) | Control (n = 275) | |||
---|---|---|---|---|---|---|---|
Categorical variables
| Maternal education | Low | 31 (35.6%)*** | 6 (23.1%) | 10 (27.8%)* | 4 (21.1%) | 37 (13.5%) |
Medium | 37 (42.5%) | 11 (42.3%) | 10 (27.8%) | 7 (36.8%) | 139 (50.5%) | ||
High | 19 (21.8%) | 9 (34.6%) | 16 (44.4%) | 8 (42.1%) | 99 (36.0%) | ||
Greek origin | 82 (93.2%) | 26 (100.0%) | 35 (97.2%) | 19 (100.0%) | 258 (94.2%) | ||
Multiparity | 58 (65.9%) | 19 (73.1%) | 20 (55.6%) | 12 (63.2%) | 187 (68.0%) | ||
Smoking during pregnancy | 19 (22.9%) | 6 (24.0%) | 10 (27.8%) | 8 (42.1%)* | 55 (20.4%) | ||
Pre-pregnancy BMI | Underweight | 6 (7.2%)*** | 1 (4.0%) | 1 (2.8%)* | 1 (5.3%) | 6 (2.2%) | |
Normal | 49 (59.0%) | 10 (40.0%) | 27 (75.0%) | 15 (78.9%) | 181 (66.8%) | ||
Overweight | 16 (19.3%) | 8 (32.0%) | 4 (11.1%) | 2 (10.5%) | 54 (19.9%) | ||
Obese | 12 (14.5%) | 6 (24.0%) | 4 (11.1%) | 1 (5.3%) | 30 (11.1%) | ||
FGR | 3 (4.0%) | 3 (14.3%) | 36 (100.0%)*** | 17 (94.4%)*** | 0 (0.0%) | ||
Continuous variables
| Maternal age, years | 29.0 (26.0 to 33.0) | 31.0 (27.2 to 36.0) | 30.0 (27.0 to 33.2) | 30.0 (28.0 to 33.0) | 31.0 (27.0 to 34.5) | |
Gestational age, weeks | 35.5 (35.0 to 36.0)*** | 36.0 (35.5 to 36.0)*** | 39.0 (37.5 to 40.0) | 38.0 (38.0 to 40.0) | 39.0 (38.0 to 39.0) | ||
Birth weight, g | 2715 (2430 to 2980)*** | 2800 (2570 to 2890)*** | 2610 (2482 to 2802)*** | 2550 (2182 to 2615)*** | 3250 (3010 to 3550) | ||
Cholesterol (n = 227) | 215.0 (189.5 to 237.0) | 222.0 (212.8 to 233.0)* | 205.0 (163.0 to 225.0) | 225.0 (198.0 to 234.0) | 202.0 (178.5 to 231.0) | ||
Triglycerides (n = 227) | 112.0 (86.5 to 134.5) | 149.0 (104.0 to 159.2) | 99.0 (89.0 to 119.0) | 95.0 (89.0 to 131.0) | 111.0 (85.5 to 138.0) | ||
Insulin (n = 369) | 6.3 (2.3 to 14.8) | 10.6 (5.0 to 17.5)* | 8.3 (3.2 to 26.6) | 5.1 (2.7 to 37.9) | 5.5 (2.0 to 15.9) | ||
LDL-C (n = 227) | 128.0 (101.5 to 138.5) | 130.0 (121.2 to 135.8) | 116.0 (90.0 to 142.0) | 142.0 (112.0 to 149.0) | 114.0 (98.5 to 142.0) | ||
HDL-C (n = 227) | 61.0 (52.0 to 71.0) | 70.5 (59.8 to 79.2) | 59.0 (49.0 to 69.0) | 63.0 (49.0 to 67.0) | 60.0 (49.0 to 68.5) | ||
Systolic BP (n = 338) | 107.7 (101.0 to 115.7) | 110.7 (105.7 to 117.3) | 105.0 (96.0 to 112.1) | 99.7 (94.2 to 110.2) | 106.3 (100.3 to 112.0) | ||
Diastolic BP (n = 338) | 69.7 (64.3 to 76.0) | 74.5 (69.6 to 79.3) | 69.3 (61.3 to 77.0) | 65.2 (58.2 to 77.3) | 69.7 (63.7 to 76.0) |
Metabolomic analysis
Selected resonance (δ), ppm
| All PB types (n = 114) | SPB (n = 88) | IPB (n = 26) | FGR (n = 36) | SGA (n = 19) | |||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Pvalue | q value | % diff |
Pvalue | q value | % diff |
Pvalue | q value | % diff |
Pvalue | q value | % diff |
Pvalue | q value | % diff | ||
Tyrosinea
| 6.87 (d), 7.18 (d) | 0.055 | 0.264 | −15% | 0.161 | 0.410 | −12% | 0.083 | 0.530 | −20% |
0.008
|
0.024
|
−24%
| 0.094 | 0.220 | −25% |
Steroid conjugate – 0.63 (s) | 0.63 (s) |
0.039
|
0.252
|
17%
|
0.045
|
0.218
|
19%
| 0.391 | 0.808 | 11% | 0.129 | 0.096 | 15% | 0.392 | 0.486 | 8% |
Leucine | 0.96 (t) | 0.364 | 0.578 | 2% | 0.227 | 0.463 | 2% | 0.778 | 0.893 | −2% | 0.443 | 0.226 | 1% |
0.026
|
0.093
|
11%
|
Lactate | 1.33 (d) | 0.342 | 0.562 | −5% | 0.259 | 0.482 | −5% | 0.988 | 0.914 | −4% |
0.025
|
0.051
|
−19%
|
0.009
|
0.081
|
−19%
|
Alanine | 1.48 (d) | 0.627 | 0.702 | −1% | 0.426 | 0.602 | −2% | 0.653 | 0.875 | 3% |
0.031
|
0.055
|
−10%
|
0.027
|
0.094
|
−9%
|
Lysine | 1.73 (m) | 0.056 | 0.264 | 1% |
0.016
|
0.214
|
2%
| 0.762 | 0.891 | −2% | 0.349 | 0.197 | −4% | 0.512 | 0.553 | 0% |
Acetate | 1.92 (s) | 0.423 | 0.613 | −2% | 0.885 | 0.758 | 0% | 0.106 | 0.564 | −7% |
0.003
|
0.018
|
−9%
|
0.017
|
0.081
|
−11%
|
N-acetyl glycoprotein fragments | 2.04 (s) | 0.390 | 0.594 | 3% | 0.783 | 0.735 | 0% |
0.009
|
0.231
|
9%
| 0.358 | 0.200 | −4% | 0.913 | 0.688 | −1% |
N-acetyl neuraminic acid | 2.06 (s) | 0.806 | 0.752 | 0% | 0.701 | 0.713 | 0% | 0.173 | 0.650 | 7% | 0.525 | 0.248 | 1% |
0.015
|
0.081
|
10%
|
Citrate | 2.55 (d) | 0.818 | 0.754 | −2% | 0.882 | 0.757 | −1% | 0.395 | 0.809 | −5% |
0.045
|
0.067
|
−9%
| 0.119 | 0.250 | −9% |
Trimethylamine | 2.87 (s) | 0.094 | 0.277 | −2% | 0.218 | 0.457 | −1% | 0.133 | 0.593 | −7% |
0.002
|
0.018
|
−18%
|
0.013
|
0.081
|
−17%
|
Trimethylamine-N-oxide | 3.27 (s) | 0.067 | 0.270 | −3% |
0.032
|
0.217
|
−3%
| 0.976 | 0.913 | 6% | 0.184 | 0.126 | −7% | 0.084 | 0.206 | −9% |
Glycine | 3.57 (d) | 0.103 | 0.279 | −5% |
0.049
|
0.218
|
−10%
| 0.920 | 0.908 | 8% |
0.019
|
0.044
|
−14%
|
0.008
|
0.081
|
−17%
|
Phenylacetylglutamine | 7.37 (d) | 0.071 | 0.271 | −9% | 0.356 | 0.558 | −3% |
0.015
|
0.231
|
−21%
| 0.871 | 0.339 | −1% | 0.428 | 0.508 | −12% |
N-methyl-2-pyridone-5-carboxamide | 8.33 (s) | 0.065 | 0.269 | 8% |
0.049
|
0.218
|
8%
| 0.676 | 0.879 | −3% | 0.764 | 0.311 | −7% | 0.545 | 0.568 | −10% |
Formate | 8.46 (s) |
0.004
|
0.105
|
−11%
|
0.009
|
0.214
|
−12%
| 0.115 | 0.574 | −8% |
0.007
|
0.024
|
−16%
| 0.410 | 0.498 | −2% |
Outcome | Metabolite | Q1 | Q2 | Q3 | Q4 |
Pvalue for trend |
---|---|---|---|---|---|---|
All PB types (n = 114) | Steroid conjugate: 0.63 (s) |
21%
|
22%
|
25%
|
32%
|
0.01*
|
Formate |
32%
|
25%
|
23%
|
20%
|
0.02*
| |
SPB (n = 88) | Steroid conjugate: 0.63 (s) |
20%
|
22%
|
24%
|
34%
|
0.01*
|
Lysine |
14%
|
30%
|
26%
|
31%
|
0.03*
| |
Trimethylamine-N-oxide | 26% | 28% | 28% | 17% | 0.10 | |
Glycine | 26% | 34% | 20% | 19% | 0.07 | |
N-methyl-2-pyridone-5-carboxamide |
19%
|
19%
|
30%
|
32%
|
0.03*
| |
Formate |
32%
|
25%
|
24%
|
19%
|
0.02
| |
IPB (n = 26) |
N-acetyl glycoprotein fragments |
8%
|
23%
|
19%
|
50%
|
0.01**
|
Phenylacetylglutamine |
42%
|
27%
|
12%
|
19%
|
0.03*
| |
FGR (n = 36) | Tyrosine |
33%
|
36%
|
17%
|
14%
|
0.03*
|
Lactate |
33%
|
33%
|
17%
|
17%
|
0.05*
| |
Alanine |
39%
|
31%
|
11%
|
19%
|
0.03*
| |
Acetate |
39%
|
31%
|
22%
|
8%
|
0.004**
| |
Citrate |
33%
|
39%
|
14%
|
14%
|
0.02*
| |
Trimethylamine |
50%
|
19%
|
17%
|
14%
|
0.002**
| |
Glycine |
33%
|
39%
|
11%
|
17%
|
0.03*
| |
Formate |
39%
|
28%
|
19%
|
14%
|
0.02*
| |
SGA (n = 19) | Leucine | 16% | 16% | 26% | 42% | 0.06 |
Lactate |
37%
|
37%
|
16%
|
11%
|
0.05*
| |
Alanine |
42%
|
32%
|
16%
|
11%
|
0.03*
| |
Acetate |
42%
|
21%
|
32%
|
5%
|
0.05*
| |
N-acetyl neuraminic acid |
21%
|
11%
|
16%
|
53%
|
0.04*
| |
Trimethylamine | 47% | 16% | 21% | 16% | 0.07 | |
Glycine |
42%
|
37%
|
11%
|
11%
|
0.02*
|
Outcomes | Metabolite | IQR | 95% CI |
Pvalue | |
---|---|---|---|---|---|
Min | Max | ||||
All PB types (n = 114)
| Steroid conjugate: 0.63 (s) | 1.90 | 0.99 | 3.69 | 0.054 |
Formate |
0.51
|
0.26
|
0.99
|
0.047
| |
SPB (n = 88)
| Steroid conjugate: 0.63 (s) | 1.99 | 0.94 | 4.32 | 0.076 |
Lysine |
2.79
|
1.20
|
6.98
|
0.021
| |
N-methyl-2-pyridone-5-carboxamide | 2.05 | 0.96 | 4.51 | 0.066 | |
Formate |
0.42
|
0.19
|
0.94
|
0.037
| |
IPB (n = 26)
|
N-acetyl glycoprotein fragments |
5.84
|
1.44
|
39.50
|
0.028
|
Phenylacetylglutamine | 0.37 | 0.09 | 1.28 | 0.131 | |
FGR (n = 36)
| Tyrosine |
0.27
|
0.08
|
0.81
|
0.025
|
Lactate | 0.37 | 0.12 | 1.04 | 0.069 | |
Alanine | 0.38 | 0.13 | 1.02 | 0.064 | |
Acetate |
0.18
|
0.04
|
0.60
|
0.011
| |
Citrate | 0.33 | 0.09 | 0.99 | 0.058 | |
Trimethylamine |
0.14
|
0.04
|
0.40
|
0.001
| |
Glycine | 0.36 | 0.11 | 1.02 | 0.062 | |
Formate |
0.24
|
0.07
|
0.71
|
0.014
| |
SGA (n = 19)
| Lactate | 0.20 | 0.03 | 0.89 | 0.055 |
Alanine | 0.19 | 0.03 | 0.88 | 0.055 | |
Acetate |
0.12
|
0.01
|
0.70
|
0.050
| |
N-acetyl neuraminic acid | 2.23 | 0.64 | 9.10 | 0.225 | |
Glycine | 0.19 | 0.03 | 0.88 | 0.052 |