Background
Methods
The NeuroFAST cohort: study design and sample collection
Collection of clinical, behavioral, and nutritional data
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Bulimic Investigatory Test Edinburgh (BITE): a 33-item self-report measure designed to assess symptomology of bulimia or binge eating, consisting of two subscales: the Symptom Scale, measuring the degree of symptoms, and the Severity Scale, providing an index of the severity of symptoms [26]. A Symptom Scale score of 20 or more indicates the presence of binge eating symptoms; a Severity Scale score of 10 or more indicates a high degree of severity. High scores on both scales indicate a high probability that a participant will fulfill the criteria for an eating disorder diagnosis. BITE is considered a valid and reliable questionnaire [27] and is widely used to assess binge eating symptomology and to screen for bulimic symptoms and their severity;
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Three-factor eating questionnaire (TFEQ): a self-report questionnaire, containing 18 items, widely used in eating behavior research in both OB and NW subjects [28]. Participants must rate each item using a 4-point Likert scale in which 1 is definitively true and 4 is definitively false. TFEQ was designed to assess three cognitive and behavioral domains of eating (factors): cognitive restraint (CR), uncontrolled eating (UE), and emotional eating (EE). The CR scale contains six items and refers to the conscious restriction of food intake to control body weight or promote weight loss. The UE scale contains nine items and refers to the tendency to eat more than usual because of a loss of control over intake. The EE scale contains three items and refers to overeating during dysphoric mood state. The TFEQ-18R was derived from a previous 51-item and 21-item version. The current version has shown a robust factor structure, good validity and internal reliability, and no significant ceiling or floor effects [29];
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Yale Food Addiction Scale (YFAS): a questionnaire developed by Gearhardt and colleagues to operationalize FA, including 25 items to assess signs of substance-dependence symptoms (e.g., tolerance, withdrawal, loss of control) in eating behavior over the past 12 months [11]. YFAS provides two scoring options: a symptom count version and a diagnostic version. To receive a diagnosis of FA, it is necessary to report experiencing three or more symptoms and clinically significant impairment or distress. Moreover, based on symptoms count, Gearhardt and colleagues suggested to dichotomize participants in high and low food addicted groups [12]. Consequently, participants with 3 or more symptoms were considered as highly food addicted while low food addicted participants scored two or fewer symptoms [23]. Accordingly, for the present study, OB participants were stratified into three groups: high addictive eating behavior, including participants with three or more symptoms and FA diagnosis (O_DHA); high addictive eating behavior, including participants with three or more symptoms but no FA diagnosis (O_HA); low addictive eating behavior, including participants with two or less symptoms (O_LA). The recently released YFAS 2.0 version [30] was not used in the present study because it was not available at the time of recruitment;
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Perceived Stress Scale (PSS): a 10-item, self-report questionnaire, assessing the individual experience of perceived stress over the past month [31, 32]. Participants must rate the frequency with which they have perceived situations as stressful using a 5-point Likert scale in which 0 is never and 4 is very often. Scores ranging from 27 to 40 are considered high perceived stress. PSS has been shown to be a reliable and valid measure of psychological stress.
Microbial DNA extraction
16S rRNA gene sequencing and bioinformatics
Co-abundance analysis
Shotgun metagenomic DNA sequencing and data analysis
RNA isolation, sequencing, and bioinformatics
Lipidomics analysis
Statistical analysis
Results
Study cohort description
All | NW | OB | |||||||
N | Median (p25-p75) | N | Median (p25-p75) | N | Median (p25-p75) | p | |||
Anthropometric, biochemical, and hormonal parameters | |||||||||
Age (years) | 100 | 34 (26–43) | 37 | 33 (27–41) | 63 | 38 (27–45) | 0.240 | ||
Body weight (kg) | 100 | 75.3 (62.1–84.0) | 37 | 58.0 (53.8–64.1) | 63 | 81.0 (76.0–90.0) | < 0.001 | ||
Body mass index, BMI (kg/m2) | 100 | 28.1 (22.8–32.8) | 37 | 21.6 (20.1–23.1) | 63 | 30.9 (28.3–33.9) | < 0.001 | ||
Systolic blood pressure (mmHg) | 99 | 120.0 (110.0–130.0) | 37 | 110.0 (110.0–120.0) | 62 | 120.0 (110.0–130.0) | 0.001 | ||
Diastolic blood pressure (mmHg) | 99 | 70.0 (70.0–80.0) | 37 | 70.0 (65.0–73.0) | 62 | 80.0 (70.0–80.0) | < 0.001 | ||
Waist circumference (cm) | 100 | 95.0 (84.0–103.0) | 37 | 80.0 (73.5–86.5) | 63 | 101.0 (95.0–107.0) | < 0.001 | ||
Hip circumference (cm) | 88 | 106.0 (97.3–115.0) | 37 | 96.0 (91.5–100.0) | 51 | 115.0 (107.0–121.0) | < 0.001 | ||
Waist-to-hip ratio | 88 | 0.86 (0.81–0.91) | 37 | 0.82 (0.79–0.88) | 51 | 0.89 (0.83–0.94) | < 0.001 | ||
Total cholesterol (mg/dl) | 100 | 180.0 (162.0–204.0) | 37 | 169.0 (160.0–192.0) | 63 | 191.0 (162.0–211.0) | 0.044 | ||
HDL-cholesterol (mg/dl) | 100 | 54.5 (46.3–62.8) | 37 | 59.0 (52.5–65.0) | 63 | 52.0 (44.0–59.0) | 0.003 | ||
Triglycerides (mg/dl) | 100 | 71.0 (53.3–99.8) | 37 | 57.0 (45.5–71.0) | 63 | 87.0 (59.0–110.0) | < 0.001 | ||
LDL-cholesterol (mg/dl) | 97 | 110.0 (96.5–130.5) | 37 | 105.0 (95.0–116.5) | 60 | 114.0 (97.3–140.8) | 0.141 | ||
Glycemia (mg/dl) | 100 | 86.0 (80.0–90.0) | 37 | 81.0 (79.0–85.5) | 63 | 89.0 (82.0–93.0) | < 0.001 | ||
Glycemia-AUC (mg/dl) | 98 | 12,930 (10,901–14,632) | 37 | 11,130 (10,275–12,735) | 61 | 14,085 (12,495–16,050) | < 0.001 | ||
Insulin (μU/ml) | 100 | 6.8 (4.2–12.0) | 37 | 3.7 (3.0–5.0) | 63 | 10.1 (6.0–14.0) | < 0.001 | ||
Insulin-AUC (μU/ml) | 97 | 5110 (3500–8017) | 37 | 3412 (2472–4326) | 60 | 7163 (4750–9966) | < 0.001 | ||
Glycated hemoglobin (%) | 100 | 5.2 (4.9–5.4) | 37 | 5.0 (4.8–5.2) | 63 | 5.3 (5.1–5.4) | < 0.001 | ||
Creatinine (mg/dl) | 45 | 0.73 (0.63–0.83) | 3 | 0.67 (0.58–0.73)* | 42 | 0.74 (0.64–0.84) | 0.300 | ||
Alanine transaminase, ALT (mg/dl) | 46 | 17.5 (12.0–24.0) | 3 | 7.0 (6.0–24.0)* | 43 | 18.0 (13.0–24.0) | 0.163 | ||
Aspartate transaminase, AST (mg/dl) | 46 | 16.0 (14.0–20.0) | 3 | 14.0 (13.0–23.0)* | 43 | 16.0 (14.0–20.0) | 0.643 | ||
Uric acid (mg/dl) | 43 | 4.3 (3.7–4.8) | 3 | 3.4 (2.8–3.9)* | 40 | 4.3 (3.9–4.9) | 0.020 | ||
Erythrosedimentation rate (mm/h) | 44 | 11.5 (6.0–19.8) | 3 | 2.0 (2.0–4.0)* | 41 | 13.0 (7.0–20.0) | 0.002 | ||
C-reactive protein (mg/dl) | 43 | 0.3 (0.1–0.5) | 3 | 0.03 (0.03–0.07)* | 40 | 0.3 (0.1–0.5) | 0.004 | ||
Thyroid stimulating hormone, TSH (mUI/ml) | 48 | 1.56 (1.30–2.11) | 4 | 1.88 (1.48–2.94) | 44 | 1.53 (1.11–2.11) | 0.395 | ||
Adrenocorticotropic hormone, ACTH (pg/ml) | 45 | 17.0 (12.0–23.0) | 3 | 11.0 (11.0–17.0)* | 42 | 17.5 (12.8–23.0) | 0.122 | ||
HOMA-IR | 100 | 1.43 (0.83–2.66) | 37 | 0.74 (0.61–0.99) | 63 | 2.16 (1.34–3.25) | < 0.001 | ||
Matsuda Index | 92 | 5.75 (3.62–9.47) | 32 | 10.3 (7.88–15.08) | 60 | 4.05 (2.83–5.83) | < 0.001 | ||
N | Mean ± sd | N | Mean ± sd | N | Mean ± sd | p | |||
Psychometric parameters | |||||||||
BITE severity score | 88 | 4.48 ± 4.51 | 33 | 1.75 ± 1.27 | 55 | 6.11 ± 4.96 | < 0.001 | ||
BITE symptom score | 88 | 6.53 ± 5.85 | 33 | 5.39 ± 3.43 | 55 | 7.21 ± 6.85 | 0.158 | ||
TFEQ UE | 90 | 16.94 ± 5.39 | 33 | 15.97 ± 4.93 | 57 | 17.51 ± 5.61 | 0.193 | ||
TFEQ CR | 90 | 14.35 ± 4.29 | 33 | 12.24 ± 2.90 | 57 | 15.58 ± 4.50 | < 0.001 | ||
TFEQ EE | 90 | 7.56 ± 2.99 | 33 | 5.73 ± 2.22 | 57 | 8.65 ± 2.87 | < 0.001 | ||
YFAS symptoms | 100 | 2.28 ± 1.55 | 37 | 1.08 ± 0.54 | 63 | 2.98 ± 1.52 | < 0.001 | ||
YFAS diagnosed addiction (a) | 14 | 14% | 37 | 0 | 63 | 14 | < 0.001 | ||
PSS | 96 | 15.53 ± 6.39 | 35 | 14.2 ± 5.19 | 61 | 16.29 ± 6.91 | 0.123 |
Gut microbiota profiling and cluster identification
Associations between the microbiota clusters and clinical and behavioral measures in normal-weight and overweight/obese women
Parameter | PCo1 | PCo2 | PCo3 | ||||||
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RC range | RC sd | p | RC range | RC sd | p | RC range | RC sd | p | |
Unweighted UniFrac PCoA for all women | |||||||||
BITE symptoms | 0.15971 | 0.02158 | 0.8 | -1.15304 | -0.28826 | 0.008 | -0.44090 | -0.12247 | 0.2 |
TFEQ UE | 0.12822 | 0.01732 | 0.9 | -1.11807 | -0.27951 | 0.005 | -0.50707 | -0.14085 | 0.2 |
C1 | C2 | C3 | C4 | p | |||||
N | Median (p25-p75) | N | Median (p25-p75) | N | Median (p25-p75) | N | Median (p25-p75) | ||
Anthropometric, biochemical, and hormonal parameters | |||||||||
Age (years) | 31 | 34 (26–42) | 33 | 32 (26–44) | 22 | 40 (30–45) | 14 | 32 (27–41) | 0.959 |
Body weight (kg) | 31 | 70.5 (58.0–80.0) | 33 | 77.8 (65.5–89.5) | 22 | 76.5 (57.5–85.5) | 14 | 76.5 (59.6–89.4) | 0.190 |
Body mass index, BMI (kg/m2) | 31 | 25.6 (21.2–30.5) | 33 | 29.7 (24.4–33.0) | 22 | 28.2 (21.8–31.8) | 14 | 28.6 (22.6–32.9) | 0.160 |
Systolic blood pressure (mmHg) | 31 | 115.0 (110.0–125.0) | 32 | 120.0 (111.3–130.0) | 22 | 112.5 (108.8–130.0) | 14 | 120.0 (110.0–130.0) | 0.287 |
Diastolic blood pressure (mmHg) | 31 | 70.0 (70.0–80.0) | 32 | 77.5 (70.0–80.0) | 22 | 70.0 (65.0–75.0) | 14 | 80.0 (70.0–81.3) | 0.150 |
Waist circumference (cm) | 31 | 88.0 (77.0–106.0) | 33 | 96.0 (89.0–102.5) | 22 | 95.5 (76.8–104.0) | 14 | 94.0 (83.0–101.3) | 0.258 |
Hip circumference (cm) | 28 | 105.0 (94.5–112.0) | 27 | 109.0 (101.0–117.0) | 20 | 103.0 (95.0–115.0) | 13 | 106.0 (97.0–120.0) | 0.274 |
Waist-to-hip ratio | 28 | 0.83 (0.79–0.92) | 27 | 0.87 (0.83–0.91) | 20 | 0.84 (0.81–0.90) | 13 | 0.88 (0.79–0.92) | 0.701 |
Total cholesterol (mg/dl) | 31 | 175.0 (164.0–202.0) | 33 | 170.0 (155.5–211.5) | 22 | 193.0 (168.3–208.8) | 14 | 185.5 (161.8–194.0) | 0.718 |
HDL-cholesterol (mg/dl) | 31 | 58.0 (47.0–64.0) | 33 | 53.0 (47.5–59.0) | 22 | 56.5 (45.5–59.8) | 14 | 56.5 (46.0–68.5) | 0.148 |
Triglycerides (mg/dl) | 31 | 69.0 (51.0–106.0) | 33 | 85.0 (60.5–106.0) | 22 | 70.0 (56.5–98.8) | 14 | 63.0 (47.5–91.5) | 0.077(a) |
LDL-cholesterol (mg/dl) | 30 | 105.5 (96.8–125.0) | 33 | 110.0 (91.0–131.5) | 20 | 115.0 (100.0–143.8) | 14 | 107.0 (92.8–126.0) | 0.940 |
Glycemia (mg/dl) | 31 | 85.0 (80.0–89.0) | 33 | 86.0 (81.0–91.0) | 22 | 88.0 (84.8–90.8) | 14 | 81.0 (78.5–90.3) | 0.600 |
Glycemia-AUC (mg/dl) | 31 | 12,630 (10,725–15,000) | 31 | 13,095 (10,515–14,715) | 22 | 13,148 (11,351–14,554) | 14 | 12,998 (10,890–14,738) | 0.911 |
Insulin (μU/ml) | 31 | 5.9 (3.2–13.0) | 33 | 9.0 (4.8–12.0) | 22 | 6.0 (4.1–13.0) | 14 | 6.0 (3.9–10.6) | 0.383 |
Insulin-AUC (μU/ml) | 31 | 4709 (2670–7095) | 31 | 7155 (4430–9725) | 21 | 4016 (3460–7379) | 14 | 5264 (3735–7728) | 0.054(b) |
Glycated hemoglobin (%) | 31 | 5.2 (4.9–5.4) | 33 | 5.3 (5.0–5.4) | 22 | 5.1 (4.9–5.4) | 14 | 5.2 (4.7–5.4) | 0.371 |
Creatinine (mg/dl) | 10 | 0.72 (0.63–0.85) | 20 | 0.75 (0.60–0.82) | 9 | 0.77 (0.63–0.90) | 6 | 0.71 (0.67–0.77) | 0.917 |
Alanine transaminase, ALT (mg/dl) | 10 | 19.0 (14.3–24.0) | 21 | 18.0 (12.0–24.5) | 9 | 17.0 (13.5–28.5) | 6 | 14.0 (9.8–17.0) | 0.180 |
Aspartate transaminase, AST (mg/dl) | 10 | 17.0 (14.8–22.3) | 21 | 16.0 (13.0–20.0) | 9 | 20.0 (15.0–23.5) | 6 | 15.0 (12.3–17.0) | 0.427 |
Uric acid (mg/dl) | 10 | 4.4 (3.7–4.8) | 21 | 4.4 (4.0–5.1) | 7 | 3.9 (3.6–4.3) | 5 | 3.5 (3.4–3.9) | 0.023(a) |
C-reactive protein | 9 | 0.4 (0.1–0.5) | 21 | 0.3 (0.1–0.5) | 8 | 0.2 (0.1–0.4) | 5 | 0.8 (0.5–0.23) | 0.253 |
Thyroid stimulating hormone, TSH (mUI/ml) | 10 | 1.43 (0.37–2.11) | 20 | 1.85 (1.51–2.38) | 10 | 1.48 (1.22–1.78) | 7 | 0.84 (0.80–1.59) | 0.059(a) |
Adrenocorticotropic hormone, ACTH (pg/ml) | 10 | 21.0 (11.0–29.3) | 21 | 17.0 (12.5–23.0) | 8 | 16.5 (13.0–23.8) | 6 | 14.5 (11.5–22.0) | 0.771 |
HOMA-IR | 31 | 1.24 (0.66–2.60) | 33 | 1.91 (0.97–2.71) | 22 | 1.37 (0.83–2.96) | 14 | 1.17 (0.77–2.37) | 0.466 |
MATSUDA Index | 29 | 6.90 (3.55–10.10) | 30 | 5.15 (2.90–7.93) | 20 | 7.05 (3.83–10.35) | 13 | 5.20 (3.90–9.45) | 0.442 |
N | Mean ± sd | N | Mean ± sd | N | Mean ± sd | N | Mean ± sd | ||
Psychometric parameters | |||||||||
BITE severity score | 30 | 4.73 ± 4.56 | 30 | 4.87 ± 5.24 | 19 | 4.31 ± 4.16 | 9 | 2.66 ± 1.73 | 0.124 |
BITE symptom score | 30 | 6.23 ± 5.63 | 30 | 5.40 ± 5.66 | 19 | 7.26 ± 6.64 | 9 | 9.77 ± 4.87 | 0.108 |
TFEQ UE | 29 | 17.10 ± 5.55 | 30 | 16.40 ± 5.87 | 19 | 16.95 ± 4.73 | 12 | 17.82 ± 5.21 | 0.254 |
TFEQ CR | 29 | 15.31 ± 5.30 | 30 | 14.36 ± 3.79 | 19 | 13.16 ± 2.77 | 12 | 13.92 ± 4.66 | 0.448 |
TFEQ EE | 29 | 7.06 ± 1.49 | 30 | 7.80 ± 2.98 | 19 | 7.42 ± 3.48 | 12 | 8.50 ± 2.35 | 0.198 |
YFAS symptoms | 31 | 1.93 ± 1.41 | 33 | 2.36 ± 1.56 | 22 | 2.41 ± 1.47 | 14 | 2.64 ± 1.95 | 0.968 |
YFAS diagnosed addiction | 2 | 6.4% | 6 | 18.2% | 4 | 18.2% | 2 | 14.3% | 0.242 |
PSS | 31 | 14.77 ± 6.27 | 32 | 16.06 ± 6.74 | 21 | 14.81 ± 6.88 | 12 | 17.33 ± 4.94 | 0.380 |