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The purpose of this investigation is to test the hypothesis that in humans, eating a relatively big amount of chocolate at the wrong time (bedtime) may disrupt our circadian system (change the circadian phase), while taking this same amount of chocolate in the morning (wake up condition) may synchronize it. Other related factors may be also affected such as total body weight and body fat, dietary habits (total energy intake and macronutrient distribution), the timing of food intake and of sleep, daily rhythms of TAP, microflora composition and postprandial glycemia.
Recent studies suggest that not only "what" the people eat, but also "when" the people eat may have a significant role in obesity treatment and in the regulation of the circadian system. Thus, the hypothesis of this study is eating a relatively big amount of chocolate at the wrong time, bedtime may affect:
Other related factors may be also affected such as total body weight and body fat, dietary habits (total energy intake and macronutrient distribution), the timing of food intake and of sleep, daily rhythms of TAP, microflora composition and postprandial glycemia.
19 women (postmenopausal) following the habitual dietary habits of participants (ad libitum) will have 30% of the habitual total daily calories in chocolate of participants (Nestle, "chocolate with milk") during two consecutive weeks each under three conditions: eating chocolate within 1 hour of habitual wake-time, eating chocolate within 1 hours of habitual bedtime, or eating no chocolate. No other chocolate (i.e., none at all in control and in the washout weeks).
The protocol will be a randomized, cross-over design, with a 1-week washout between each condition.
During the 14 days in each condition, the participants will record sleep and activity schedules by dairy, food intake and food timing by phone application, daily rhythms of wrist temperature, activity and position (TAP).
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| Label | Type | Description | Intervention Names |
|---|---|---|---|
| Morning Chocolate, then Evening/Night Chocolate and Control Chocolate | Experimental | First condition is eating 100gr chocolate within one hour of habitual waketime. Second condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Third condition is eating no milk chocolate or other chocolate. |
|
| Morning Chocolate, then Control Chocolate and Evening/Night Chocolate | Experimental | First condition is eating 100gr chocolate within one hour of habitual waketime. Second condition is eating no milk chocolate or other chocolate. Third condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. |
|
| Evening/Night Chocolate, then Morning Chocolate and Control Chocolate | Experimental | First condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Second condition is eating 100gr chocolate within one hour of habitual waketime. Third condition is eating no milk chocolate or other chocolate. |
|
| Evening/Night Chocolate, then Control Chocolate and Morning Chocolate | Experimental | First condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Second condition is eating no milk chocolate or other chocolate. Third condition is eating 100gr chocolate within one hour of habitual waketime. |
| Name | Type | Description | Arm Group Labels | Other Names |
|---|---|---|---|---|
| Chocolate 100gr | Behavioral | Evening/Night Chocolate: Eating 100gr of milk chocolate within 1 hours of habitual bedtime Control Chocolate: Eating no milk chocolate or other chocolate Chocolate Morning: Eating chocolate within one hour of habitual waketime. |
| Measure | Description | Time Frame |
|---|---|---|
| Resting Energy Expenditure (Indirect Calorimetry) | The investigators will measure by indirect calorimetry the resting energy expenditure after each condition (no chocolate, Morning Chocolate, Night Chocolate) Changes in Resting Energy Expenditure (Indirect Calorimetry) were determined by changes in resting energy expenditure occurred in the participants between baseline and after 2 weeks in each of the three different experimental conditions. Oxygen (O2) consumption (mL/min) and carbon dioxide (CO2) production (mL/min) were measured. The Respiratory Quotient (RQ) was calculated out of O2 consumption (mL/min) and CO2 production (mL/min). Energy expenditure values (kcal/day) were calculated according to the Weir equation: Metabolic rate (kcal per day) = 1440 (3.9 VO2 + 1.1 VCO2)* | Baseline and after 2 weeks in each condition |
| Total Body Weight | The investigators will measure total body weight after each condition (no chocolate, Morning Chocolate, Night Chocolate) | Baseline and after 2 weeks in each condition |
| Measure | Description | Time Frame |
|---|---|---|
| Fragmentation of Wrist Temperature (WT) Daily Rhythm | The investigators will measure the fragmentation parameter derived from wrist temperature after each condition (no chocolate, Morning Chocolate, Night Chocolate). Intradaily variability (IV) characterizes the rhythm fragmentation. Changes in daily rhythm of wrist temperature fragmentation were measured by changes in the fragmentation parameter derived from wrist temperature along the two weeks of intervention in each of the three conditions. The participants wore a wristwatch during the last 7 days of each condition on the non-dominant wrist to record body temperature and activity. Wrist temperature parameters were measured to represent the wrist body temperature pattern, which provides an objectively measure of sleep and siesta (together with activity rhythm). A low fragmentation involves a low intraday rhythmicity of the circadian rhythm. Its values oscillated between 0, when the wave was perfectly sinusoidal, and 2, when the wave was as Gaussian noise. |
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Exclusion Criteria:
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| PubMed Identifier | Type | Citation | Retractions |
|---|---|---|---|
| 34164846 | Derived | Hernandez-Gonzalez T, Gonzalez-Barrio R, Escobar C, Madrid JA, Periago MJ, Collado MC, Scheer FAJL, Garaulet M. Timing of chocolate intake affects hunger, substrate oxidation, and microbiota: A randomized controlled trial. FASEB J. 2021 Jul;35(7):e21649. doi: 10.1096/fj.202002770RR. |
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| ID | Title | Description |
|---|---|---|
| FG000 | Morning Chocolate, Then Evening/Night Chocolate and Control Chocolate | First condition is eating 100gr chocolate within one hour of habitual waketime. Second condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime Third condition is eating no milk chocolate or other chocolate |
| FG001 | Morning Chocolate, Then Control Chocolate and Evening/Night Chocolate | First condition is eating 100gr chocolate within one hour of habitual waketime. Second condition is eating no milk chocolate or other chocolate Third condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime |
| FG002 | Evening/Night Chocolate, Then Morning Chocolate and Control Chocolate | First condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime Second condition is eating 100gr chocolate within one hour of habitual waketime. Third condition is eating no milk chocolate or other chocolate |
| FG003 | Evening/Night Chocolate, Then Control Chocolate and Morning Chocolate | First condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime Second condition is eating no milk chocolate or other chocolate Third condition is eating 100gr chocolate within one hour of habitual waketime. |
| FG004 | Control Chocolate, Then Morning Chocolate and Evening/Night Chocolate | First condition is eating no milk chocolate or other chocolate Second condition is eating 100gr chocolate within one hour of habitual waketime. Third condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime |
| FG005 | Control Chocolate, Then Evening/Night Chocolate and Morning Chocolate | First condition is eating no milk chocolate or other chocolate Second condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime Third condition is eating 100gr chocolate within one hour of habitual waketime. |
| Title | Milestones | Reasons Not Completed | ||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| First Intervention (2 Weeks) |
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| Washsout (1 Week) |
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| Second Intervention (2 Weeks) |
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| Washsout (1 Week) |
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| Third Intervention (2 Weeks) |
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| ID | Title | Description |
|---|---|---|
| BG000 | Morning Chocolate, Then Evening/Night Chocolate and Control Chocolate | First condition is eating 100gr chocolate within one hour of habitual waketime. Second condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime Third condition is eating no milk chocolate or other chocolate. |
| BG001 |
| Units | Counts |
|---|---|
| Participants |
|
| Title | Description | Population Description | Parameter Type | Dispersion Type | Unit of Measure | Calculate Percentage | Denominator Units Selected | Denominators | Classes |
|---|---|---|---|---|---|---|---|---|---|
| Age, Customized | Mean |
| Type | Title | Description | Population Description | Reporting Status | Anticipated Posting Date | Parameter Type | Dispersion Type | Unit of Measure | Calculate Percentage | Time Frame | Units Analyzed | Denominator Units Selected | Arm/Group Information | Denominators | Classes | Analyses | |||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Primary | Resting Energy Expenditure (Indirect Calorimetry) | The investigators will measure by indirect calorimetry the resting energy expenditure after each condition (no chocolate, Morning Chocolate, Night Chocolate) Changes in Resting Energy Expenditure (Indirect Calorimetry) were determined by changes in resting energy expenditure occurred in the participants between baseline and after 2 weeks in each of the three different experimental conditions. Oxygen (O2) consumption (mL/min) and carbon dioxide (CO2) production (mL/min) were measured. The Respiratory Quotient (RQ) was calculated out of O2 consumption (mL/min) and CO2 production (mL/min). Energy expenditure values (kcal/day) were calculated according to the Weir equation: Metabolic rate (kcal per day) = 1440 (3.9 VO2 + 1.1 VCO2)* | Posted | Mean | Standard Deviation | kcal/day | Baseline and after 2 weeks in each condition |
|
Adverse events were collected for each arm in the three conditions (Morning, Evening/Night, Control) during two weeks each, along with washout periods of one week between conditions. In total, adverse events were reported for up to eight weeks.
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| ID | Title | Description | Deaths (Affected) | Deaths (At Risk) | Serious Events (Affected) | Serious Events (At Risk) | Other Events (Affected) | Other Events (At Risk) |
|---|---|---|---|---|---|---|---|---|
| EG000 | Morning Chocolate First, Second Evening/Night Chocolate and Control Chocolate: Morning Condition | The intervention is eating chocolate within one hour of habitual waketime. Evening/Night Chocolate and Control Chocolate: Behavioral: Evening/Night Chocolate Eating 100gr of milk chocolate within (1 hours of habitual bedtime) Behavioral: Control Chocolate Eating no milk chocolate or other chocolate |
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| Title | Organization | Phone | Extension | |
|---|---|---|---|---|
| Prof. Marta Garaulet Aza | Universidad de Murcia | +34868889478 | garaulet@um.es |
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| Control Chocolate, then Morning Chocolate and Evening/Night Chocolate | Experimental | First condition is eating no milk chocolate or other chocolate. Second condition is eating 100gr chocolate within one hour of habitual waketime. Third condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. |
|
| Control Chocolate, then Evening/Night Chocolate and Morning Chocolate | Experimental | First condition is eating no milk chocolate or other chocolate. Second condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Third condition is eating 100gr chocolate within one hour of habitual waketime. |
|
| Baseline and after 2 weeks in each condition |
| Regularity of Wrist Temperature (WT) Daily Rhythm | The investigators will measure the regularity parameter derived from wrist temperature after each condition (no chocolate, Morning Chocolate, Night Chocolate). RegularIty is measured as interdaily stability (IS), that means rhythm stability over different days; it varied between 0 for Gaussian noise to 1 for perfect stability, where the rhythm repeated itself exactly day after day. Changes in daily rhythm of wrist temperature regularity were measured by changes in the regularity parameter derived from wrist temperature along the two weeks of intervention in each of the three conditions. A regular rhythm means recurring at uniform intervals. The participants wore a wristwatch during the last 7 days of each condition on the non-dominant wrist to record body temperature and activity. Wrist temperature parameters were measured to represent the wrist body temperature pattern, which provides an objectively measure of sleep and siesta (together with activity rhythm). | Baseline and after 2 weeks in each condition |
| Amplitude of Wrist Temperature (WT) Daily Rhythm | The investigators will measure the amplitude parameter derived from wrist temperature after each condition (no chocolate, Morning Chocolate, Night Chocolate). In the cosinor analysis to characterize the WT rhythm, we calculate amplitude (difference between the maximum [or minimum] value of the cosine function and mesor) Changes in daily rhythm of wrist temperature amplitude were measured by changes in the amplitude parameter derived from wrist temperature along the two weeks of intervention in each of the three conditions. The participants wore a wristwatch during the last 7 days of each condition on the non-dominant wrist to record body temperature and activity. Wrist temperature parameters were measured to represent the wrist body temperature pattern, which provides an objectively measure of sleep and siesta (together with activity rhythm). A healthy circadian rhythm is considered as the one with high amplitude. | Baseline and after 2 weeks in each condition |
| Changes in Microbiota Diversity (Inverse Simpson Index) | Microbiota diversity changes were measured using Inverse Simpson Index, accounting for both species richness (number of species) and evenness (distribution of species). A higher Inverse Simpson Index indicates greater diversity because it reduces the impact of dominant species and gives more weight to rarer species. Unlike the regular Simpson Index (which decreases as diversity increases), the inverse form is more intuitive: higher values mean a more diverse microbiota. This index is widely used in microbiome studies to compare microbial diversity across different samples, such as in health versus disease states. | Baseline and after 2 weeks in each condition |
| Concentration of Total Short-chain Fatty Acids (SCFAs) | Short-chain fatty acids (SCFAs) in fecal samples were analyzed using gas-liquid chromatography (GLC) with a GC-FID system. Samples were mixed with NaOH, lyophilized, and homogenized with formic acid, methanol, and an internal standard (2-ethyl butyric acid). After ultrasonic treatment and centrifugation, the supernatant was filtered and injected into an Agilent 7890A GC system equipped with a Nukol capillary column. Helium was used as the carrier gas, and the FID temperature was set at 220°C. SCFAs were identified by comparing retention times with authentic standards, and quantification was performed using calibration curves with a high correlation (R² = 0.99). Total average values of SCFA (acetate, propionate, other minorities) in each condition were calculated. | After 2 weeks in each condition |
| Energy Intake | The investigators will measure changes in total energy intake measured as kilocalories between baseline and after 2 weeks in each condition (no chocolate, Morning Chocolate, Night Chocolate) At baseline and during the 2 weeks of each condition, time-stamped photographs were captured with a cell phone app. Total energy intake during the 14 days of each condition was analyzed with the nutritional evaluation software (Grunumur 2.0 8). | Baseline and after 2 weeks in each condition |
| Macronutrient Composition | The investigators will measure macronutrient composition measured as and grams per day between baseline and after 2 weeks in each condition (no chocolate, Morning Chocolate, Night Chocolate). At baseline and during the 2 weeks of each condition, time-stamped photographs were captured with a cell phone app. Macronutrient composition during the 14 days of each condition was analyzed with the nutritional evaluation software (Grunumur 2.0 8). | Baseline and after 2 weeks in each condition |
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| NOT COMPLETED |
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| COMPLETED |
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| NOT COMPLETED |
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| COMPLETED |
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| NOT COMPLETED |
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| COMPLETED |
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| Morning Chocolate, Then Control Chocolate and Evening/Night Chocolate |
First condition is eating 100gr chocolate within one hour of habitual waketime. Second condition is eating no milk chocolate or other chocolate Third condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. |
| BG002 | Evening/Night Chocolate, Then Morning Chocolate and Control Chocolate | First condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime Second condition is eating 100gr chocolate within one hour of habitual waketime. |
| BG003 | Evening/Night Chocolate, Then Control Chocolate and Morning Chocolate | First condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime Second condition is eating no milk chocolate or other chocolate Third condition is eating 100gr chocolate within one hour of habitual waketime. |
| BG004 | Control Chocolate, Then Morning Chocolate and Evening/Night Chocolate | First condition is eating no milk chocolate or other chocolate Second condition is eating 100gr chocolate within one hour of habitual waketime. |
| BG005 | Control Chocolate, Then Evening/Night Chocolate and Morning Chocolate | First condition is eating no milk chocolate or other chocolate Second condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime Third condition is eating 100gr chocolate within one hour of habitual waketime. |
| BG006 | Total | Total of all reporting groups |
| years |
|
| Sex: Female, Male | Count of Participants | Participants | No |
|
First condition is eating 100gr chocolate within one hour of habitual waketime. Second condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime.
Third condition is eating no milk chocolate or other chocolate.
| OG001 | Morning Chocolate, Then Control Chocolate and Evening/Night Chocolate | First condition is eating 100gr chocolate within one hour of habitual waketime. Second condition is eating no milk chocolate or other chocolate. Third condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. |
| OG002 | Evening/Night Chocolate, Then Morning Chocolate and Control Chocolate | First condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Second condition is eating 100gr chocolate within one hour of habitual waketime. Third condition is eating no milk chocolate or other chocolate |
| OG003 | Evening/Night Chocolate, Then Control Chocolate and Morning Chocolate | First condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Second condition is eating no milk chocolate or other chocolate. Third condition is eating 100gr chocolate within one hour of habitual waketime. |
| OG004 | Control Chocolate, Then Morning Chocolate and Evening/Night Chocolate | First condition is eating no milk chocolate or other chocolate. Second condition is eating 100gr chocolate within one hour of habitual waketime. Third condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime |
| OG005 | Control Chocolate, Then Evening/Night Chocolate and Morning Chocolate | First condition is eating no milk chocolate or other chocolate. Second condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Third condition is eating 100gr chocolate within one hour of habitual waketime. |
|
|
| Primary | Total Body Weight | The investigators will measure total body weight after each condition (no chocolate, Morning Chocolate, Night Chocolate) | Posted | Mean | Standard Deviation | kilograms (kg) | Baseline and after 2 weeks in each condition |
|
|
|
| Secondary | Fragmentation of Wrist Temperature (WT) Daily Rhythm | The investigators will measure the fragmentation parameter derived from wrist temperature after each condition (no chocolate, Morning Chocolate, Night Chocolate). Intradaily variability (IV) characterizes the rhythm fragmentation. Changes in daily rhythm of wrist temperature fragmentation were measured by changes in the fragmentation parameter derived from wrist temperature along the two weeks of intervention in each of the three conditions. The participants wore a wristwatch during the last 7 days of each condition on the non-dominant wrist to record body temperature and activity. Wrist temperature parameters were measured to represent the wrist body temperature pattern, which provides an objectively measure of sleep and siesta (together with activity rhythm). A low fragmentation involves a low intraday rhythmicity of the circadian rhythm. Its values oscillated between 0, when the wave was perfectly sinusoidal, and 2, when the wave was as Gaussian noise. | Posted | Mean | Standard Deviation | arbitrary units | Baseline and after 2 weeks in each condition |
|
|
|
| Secondary | Regularity of Wrist Temperature (WT) Daily Rhythm | The investigators will measure the regularity parameter derived from wrist temperature after each condition (no chocolate, Morning Chocolate, Night Chocolate). RegularIty is measured as interdaily stability (IS), that means rhythm stability over different days; it varied between 0 for Gaussian noise to 1 for perfect stability, where the rhythm repeated itself exactly day after day. Changes in daily rhythm of wrist temperature regularity were measured by changes in the regularity parameter derived from wrist temperature along the two weeks of intervention in each of the three conditions. A regular rhythm means recurring at uniform intervals. The participants wore a wristwatch during the last 7 days of each condition on the non-dominant wrist to record body temperature and activity. Wrist temperature parameters were measured to represent the wrist body temperature pattern, which provides an objectively measure of sleep and siesta (together with activity rhythm). | Posted | Mean | Standard Deviation | arbitrary units | Baseline and after 2 weeks in each condition |
|
|
|
| Secondary | Amplitude of Wrist Temperature (WT) Daily Rhythm | The investigators will measure the amplitude parameter derived from wrist temperature after each condition (no chocolate, Morning Chocolate, Night Chocolate). In the cosinor analysis to characterize the WT rhythm, we calculate amplitude (difference between the maximum [or minimum] value of the cosine function and mesor) Changes in daily rhythm of wrist temperature amplitude were measured by changes in the amplitude parameter derived from wrist temperature along the two weeks of intervention in each of the three conditions. The participants wore a wristwatch during the last 7 days of each condition on the non-dominant wrist to record body temperature and activity. Wrist temperature parameters were measured to represent the wrist body temperature pattern, which provides an objectively measure of sleep and siesta (together with activity rhythm). A healthy circadian rhythm is considered as the one with high amplitude. | Posted | Mean | Standard Deviation | ºC | Baseline and after 2 weeks in each condition |
|
|
|
| Secondary | Changes in Microbiota Diversity (Inverse Simpson Index) | Microbiota diversity changes were measured using Inverse Simpson Index, accounting for both species richness (number of species) and evenness (distribution of species). A higher Inverse Simpson Index indicates greater diversity because it reduces the impact of dominant species and gives more weight to rarer species. Unlike the regular Simpson Index (which decreases as diversity increases), the inverse form is more intuitive: higher values mean a more diverse microbiota. This index is widely used in microbiome studies to compare microbial diversity across different samples, such as in health versus disease states. | Posted | Mean | Standard Deviation | Index | Baseline and after 2 weeks in each condition |
|
|
|
| Secondary | Concentration of Total Short-chain Fatty Acids (SCFAs) | Short-chain fatty acids (SCFAs) in fecal samples were analyzed using gas-liquid chromatography (GLC) with a GC-FID system. Samples were mixed with NaOH, lyophilized, and homogenized with formic acid, methanol, and an internal standard (2-ethyl butyric acid). After ultrasonic treatment and centrifugation, the supernatant was filtered and injected into an Agilent 7890A GC system equipped with a Nukol capillary column. Helium was used as the carrier gas, and the FID temperature was set at 220°C. SCFAs were identified by comparing retention times with authentic standards, and quantification was performed using calibration curves with a high correlation (R² = 0.99). Total average values of SCFA (acetate, propionate, other minorities) in each condition were calculated. | Posted | Mean | Standard Deviation | mmol/mg feces | After 2 weeks in each condition |
|
|
|
| Secondary | Energy Intake | The investigators will measure changes in total energy intake measured as kilocalories between baseline and after 2 weeks in each condition (no chocolate, Morning Chocolate, Night Chocolate) At baseline and during the 2 weeks of each condition, time-stamped photographs were captured with a cell phone app. Total energy intake during the 14 days of each condition was analyzed with the nutritional evaluation software (Grunumur 2.0 8). | Posted | Mean | Standard Deviation | kcal | Baseline and after 2 weeks in each condition |
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|
| Secondary | Macronutrient Composition | The investigators will measure macronutrient composition measured as and grams per day between baseline and after 2 weeks in each condition (no chocolate, Morning Chocolate, Night Chocolate). At baseline and during the 2 weeks of each condition, time-stamped photographs were captured with a cell phone app. Macronutrient composition during the 14 days of each condition was analyzed with the nutritional evaluation software (Grunumur 2.0 8). | Posted | Mean | Standard Deviation | grams/day | Baseline and after 2 weeks in each condition |
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| 3 |
| 0 |
| 3 |
| 0 |
| 3 |
| EG001 | Morning Chocolate First, Second Control Chocolate and Evening/Night Chocolate: Morning Condition | The intervention is eating chocolate within one hour of habitual waketime. Control Chocolate and Evening/Night Chocolate: Behavioral: Control Chocolate Eating no milk chocolate or other chocolate Behavioral: Evening/Night Chocolate Eating 100gr of milk chocolate (within 1 hours of habitual bedtime) | 0 | 3 | 0 | 3 | 0 | 3 |
| EG002 | Evening/Night Chocolate First, Second Morning Chocolate and Control Chocolate: Morning Condition | The intervention is eating chocolate within one hour of habitual waketime. Morning Chocolate and Control Chocolate: Behavioral: Morning Chocolate Eating 100gr of milk chocolate (within 1 hours of habitual waketime) Behavioral: Control Chocolate Eating no milk chocolate or other chocolate | 0 | 5 | 0 | 5 | 0 | 5 |
| EG003 | Evening/Night Chocolate First, Second Control Chocolate and Morning Chocolate: Morning Condition | The intervention is eating chocolate within one hour of habitual waketime. Control Chocolate and Morning Chocolate: Behavioral: Control Chocolate Eating no milk chocolate or other chocolate. Behavioral: Morning Chocolate Eating 100gr of milk chocolate (within 1 hours of habitual waketime) | 0 | 2 | 0 | 2 | 0 | 2 |
| EG004 | Control Chocolate First, Second Morning Chocolate and Evening/Night Chocolate: Morning Condition | The intervention is eating chocolate within one hour of habitual waketime. Morning Chocolate and Evening/Night Chocolate: Behavioral: Morning Chocolate Eating 100gr of milk chocolate (within 1 hours of habitual waketime) Behavioral: Evening/Night Chocolate Eating 100gr of milk chocolate (within 1 hours of habitual bedtime) | 0 | 3 | 0 | 3 | 0 | 3 |
| EG005 | Control Chocolate First, Second Evening/Night Chocolate and Morning Chocolate: Morning Condition | The intervention is eating chocolate within one hour of habitual waketime. Evening/Night Chocolate and Morning Chocolate: Behavioral: Evening/Night Chocolate Eating 100gr of milk chocolate (within 1 hours of habitual bedtime) Behavioral: Morning Chocolate Eating 100gr of milk chocolate (within 1 hours of habitual waketime) | 0 | 3 | 0 | 3 | 0 | 3 |
| EG006 | Morning Chocolate First, Second Evening/Night Chocolate and Control Chocolate: Evening Condition | The intervention is eating chocolate within one hour of habitual waketime. Evening/Night Chocolate and Control Chocolate: Behavioral: Evening/Night Chocolate Eating 100gr of milk chocolate within (1 hours of habitual bedtime) Behavioral: Control Chocolate Eating no milk chocolate or other chocolate | 0 | 3 | 0 | 3 | 0 | 3 |
| EG007 | Morning Chocolate First, Second Control Chocolate and Evening/Night Chocolate: Evening Condition | The intervention is eating chocolate within one hour of habitual waketime. Control Chocolate and Evening/Night Chocolate: Behavioral: Control Chocolate Eating no milk chocolate or other chocolate Behavioral: Evening/Night Chocolate Eating 100gr of milk chocolate (within 1 hours of habitual bedtime) | 0 | 3 | 0 | 3 | 0 | 3 |
| EG008 | Evening/Night Chocolate First, Second Morning Chocolate and Control Chocolate: Evening Condition | The intervention is eating chocolate within one hour of habitual waketime. Morning Chocolate and Control Chocolate: Behavioral: Morning Chocolate Eating 100gr of milk chocolate (within 1 hours of habitual waketime) Behavioral: Control Chocolate Eating no milk chocolate or other chocolate | 0 | 5 | 0 | 5 | 0 | 5 |
| EG009 | Evening/Night Chocolate First, Second Control Chocolate and Morning Chocolate: Evening Condition | The intervention is eating chocolate within one hour of habitual waketime. Control Chocolate and Morning Chocolate: Behavioral: Control Chocolate Eating no milk chocolate or other chocolate. Behavioral: Morning Chocolate Eating 100gr of milk chocolate (within 1 hours of habitual waketime) | 0 | 2 | 0 | 2 | 0 | 2 |
| EG010 | Control Chocolate First, Then Morning Chocolate and Evening/Night Chocolate: Evening Condition | The intervention is eating chocolate within one hour of habitual waketime. Morning Chocolate and Evening/Night Chocolate: Behavioral: Morning Chocolate Eating 100gr of milk chocolate (within 1 hours of habitual waketime) Behavioral: Evening/Night Chocolate Eating 100gr of milk chocolate (within 1 hours of habitual bedtime) | 0 | 3 | 0 | 3 | 0 | 3 |
| EG011 | Control Chocolate First, Second Evening/Night Chocolate and Morning Chocolate: Evening Condition | The intervention is eating chocolate within one hour of habitual waketime. Evening/Night Chocolate and Morning Chocolate: Behavioral: Evening/Night Chocolate Eating 100gr of milk chocolate (within 1 hours of habitual bedtime) Behavioral: Morning Chocolate Eating 100gr of milk chocolate (within 1 hours of habitual waketime) | 0 | 3 | 0 | 3 | 0 | 3 |
| EG012 | Morning Chocolate First, Second Evening/Night Chocolate and Control Chocolate: Control Condition | The intervention is eating chocolate within one hour of habitual waketime. Evening/Night Chocolate and Control Chocolate: Behavioral: Evening/Night Chocolate Eating 100gr of milk chocolate within (1 hours of habitual bedtime) Behavioral: Control Chocolate Eating no milk chocolate or other chocolate | 0 | 3 | 0 | 3 | 0 | 3 |
| EG013 | Morning Chocolate First, Second Control Chocolate and Evening/Night Chocolate: Control Condition | The intervention is eating chocolate within one hour of habitual waketime. Control Chocolate and Evening/Night Chocolate: Behavioral: Control Chocolate Eating no milk chocolate or other chocolate Behavioral: Evening/Night Chocolate Eating 100gr of milk chocolate (within 1 hours of habitual bedtime) | 0 | 3 | 0 | 3 | 0 | 3 |
| EG014 | Evening/Night Chocolate First, Second Morning Chocolate and Control Chocolate: Control Condition | The intervention is eating chocolate within one hour of habitual waketime. Morning Chocolate and Control Chocolate: Behavioral: Morning Chocolate Eating 100gr of milk chocolate (within 1 hours of habitual waketime) Behavioral: Control Chocolate Eating no milk chocolate or other chocolate | 0 | 5 | 0 | 5 | 0 | 5 |
| EG015 | Evening/Night Chocolate First, Second Control Chocolate and Morning Chocolate: Control Condition | The intervention is eating chocolate within one hour of habitual waketime. Control Chocolate and Morning Chocolate: Behavioral: Control Chocolate Eating no milk chocolate or other chocolate. Behavioral: Morning Chocolate Eating 100gr of milk chocolate (within 1 hours of habitual waketime) | 0 | 2 | 0 | 2 | 0 | 2 |
| EG016 | Control Chocolate First, Second Morning Chocolate and Evening/Night Chocolate: Control Condition | The intervention is eating chocolate within one hour of habitual waketime. Morning Chocolate and Evening/Night Chocolate: Behavioral: Morning Chocolate Eating 100gr of milk chocolate (within 1 hours of habitual waketime) Behavioral: Evening/Night Chocolate Eating 100gr of milk chocolate (within 1 hours of habitual bedtime) | 0 | 3 | 0 | 3 | 0 | 3 |
| EG017 | Control Chocolate First, Second Evening/Night Chocolate and Morning Chocolate: Control Condition | The intervention is eating chocolate within one hour of habitual waketime. Evening/Night Chocolate and Morning Chocolate: Behavioral: Evening/Night Chocolate Eating 100gr of milk chocolate (within 1 hours of habitual bedtime) Behavioral: Morning Chocolate Eating 100gr of milk chocolate (within 1 hours of habitual waketime) | 0 | 3 | 0 | 3 | 0 | 3 |
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| Weight at Morning condition |
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| Weight at Night condition |
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| Weight at Control condition |
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| Fragmentation of WT rhythm at Morning condition |
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| Fragmentation of WT rhythm at Night condition |
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| Fragmentation of WT rhythm at Control condition |
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| Regularity of WT daily rhythm at Morning condition |
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| Regularity of WT daily rhythm at Night condition |
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| Regularity of WT daily rhythm at Control condition |
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| Amplitude of WT daily rhythm at Morning condition |
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| Amplitude of WT daily rhythm at Night condition |
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| Amplitude of WT daily rhythm at Control condition |
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| Inverse Simpson Index Control condition |
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| Inverse Simpson Index Morning condition |
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| Evening condition |
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| Total SCFA Morning condition |
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| Total SCFA Evening condition |
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| Energy intake at Morning condition |
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| Energy intake at Night condition |
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| Energy intake at Control condition |
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| Fat at Baseline |
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| Protein at Baseline |
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| Carbohydrates at Morning condition |
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| Fat at Morning condition |
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| Protein at Morning condition |
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| Carbohydrates at Night condition |
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| Fat at Night condition |
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| Protein at Night condition |
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| Carbohydrates at Control condition |
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| Fat at Control condition |
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| Protein at Control condition |
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