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Differential abundance
Differential abundance of VPs among VLP samples on day 2 was determined with ANCOM-BC v1.2.2
PMC10497675
Phage-host interactions
To determine the dynamics of phage-bacterium interaction across the entire population, the change in relative abundance between days 0 and 2, 2 and 28 and 0 and 28 were determined for all VPs with a host and all MAGs with a known phage. The resulting values were then averaged for both VPs and MAGs at the species level,...
PMC10497675
Patient and public involvement statement
Diabetes II, Diabetes
DIABETES
Patients were involved in the assessment of the grant proposals for this study by the Dutch Diabetes Research Foundation (Diabetes II Breakthrough grant (459001008) and Senior Fellowship (2019.82.004)). Moreover, the patient panel advised on the patient burden of the clinical study. In addition, patients were involved ...
PMC10497675
Ethics approval and informed consent statement
This study involves human participants and was approved by the Medical Research Ethics Committee Academic Medical Center Amsterdam (METC 2018_231). Both participants and faeces donors gave informed consent to participate in the study before taking part. The study was registered at the Dutch National Trial Register (NTR...
PMC10497675
Reporting summary
Further information on research design is available in the 
PMC10497675
Supplementary information
Supplementary InformationPeer Review FileDescription of Additional Supplementary FilesSupplementary Dataset 1Reporting Summary
PMC10497675
Supplementary information
The online version contains supplementary material available at 10.1038/s41467-023-41329-z.
PMC10497675
Acknowledgements
Diabetes II, Diabetes
RECRUITMENT, DIABETES
We want to thank Aline Fenneman, Klaartje de Bruin and Melanie Benard for their help with stool donor recruitment and screening. In addition, we thank Veera Houttu, Ulrika Boulund, Kim Dzobo and Torsten Scheithauer for performing the randomisation to guarantee blinding of the investigator and volunteer. We would also l...
PMC10497675
Author contributions
H.H. and M.N. conceived the research idea and designed the study; K.W. performed the clinical study; K.W. and T.P.M.S. processed the samples in the laboratory; P.A.dJ., K.W. and I.A. performed data analysis; K.W. and P.A.dJ. wrote the first draft of the manuscript; All authors contributed to manuscript revision, read a...
PMC10497675
Peer review
PMC10497675
Data availability
The sequencing data generated in this study have been deposited in the European Nucleotide Archive database under accession code:
PMC10497675
Code availability
The following programmes were used for data collection: Castor EDC (v2019.2.0-2020.2.25), EPIC (versions August 2019-November 2020), Voedingscentrum Eetmeter (v2019-2020), Freestyle Libre 1 software. The following software and codes were used for data analysis: R v4.2.1, CGDA v0.8.2, fastp v0.23.2, bbmap v38.90, metaSP...
PMC10497675
Competing interests
FOUNDER
M.N. is founder and scientific advisor of Caelus Health, however none of this bears any relevance to the content of the current paper. K.W., P.A.dJ., T.P.M.S., I.A., E.M.K. and H.H. report no conflict of interest.
PMC10497675
References
PMC10497675
1. Introduction
eating behaviors, weight loss
Multi-ingredient thermogenic supplements can acutely increase resting energy expenditure (REE) and subjective energy. However, less is understood about the effects of chronic consumption on body composition, metabolism, and subjective variables such as mood, sleep quality, and eating behaviors. Fifty-two healthy, exerc...
PMC10674526
2. Materials and Methods
PMC10674526
2.1. Participants
Generally healthy, non-smoking, exercise-trained adults between 18 and 40 years of age who had maintained a relatively stable body weight (i.e., no more than a gain or loss of approximately 4.5 kg based on self-report [
PMC10674526
2.2. Ethical Approval
This study was conducted according to the guidelines established by the Declaration of Helsinki and all procedures involving human subjects were approved by the Texas Tech University Institutional Review Board (IRB2022-993). Written informed consent was obtained from all subjects. Plans for data collection were also re...
PMC10674526
2.3. Pre-Assessment Standardization
Data collection consisted of an initial visit to collect baseline measures and a follow-up visit four weeks later for post-intervention assessments [Upon arrival at the laboratory during the initial visit, participants were screened to confirm eligibility, the investigation was verbally explained to participants, and w...
PMC10674526
2.4. Body Composition and Anthropometric Assessments
LUNG
All devices were calibrated and used in accordance with manufacturer instructions and using methods previously described in detail elsewhere [Next, a dual-energy X-ray absorptiometry (DXA; iDXA, General Electric, Boston, MA, USA with enCORE software version 16.10.151, 16 [SP 1]) scan was conducted to provide an estimat...
PMC10674526
2.5. Psychometric Questionnaires and Dietary Intake Assessment
depression, eating behavior, mood disturbance
Validated questionnaires were used to elicit data related to participants’ physical activity, mood, sleep habits, and eating behavior at both timepoints. These questionnaires included the International Physical Activity Questionnaire (IPAQ) short form [The MFQ assesses mood disturbance over the past two weeks, with a m...
PMC10674526
2.6. Resting Metabolic Rate
Resting Metabolic Rate (RMR) was assessed via indirect calorimetry using a metabolic cart (TrueOne 2400, Parvo Medics, Salt Lake City, UT, USA) with gas and flow calibration performed each day in accordance with manufacturer instructions. The testing protocol followed current best practice guidelines for the assessment...
PMC10674526
2.7. Participant Allocation
RECRUITMENT
After baseline testing was complete, participants were stratified by sex (male or female), training status (resistance-trained, endurance-trained, or concurrently trained), and body fat percentage (greater or less than 20% for males, and greater or less than 25% for females) and were allocated to an intervention using ...
PMC10674526
2.8. Intervention
BLIND
Participants allocated to CON were instructed to limit their caffeine intake to no more than approximately 100 mg per day and to refrain from the use of any performance-enhancing or thermogenic supplement for the duration of the four-week study. If participants were already using supplements for general health support ...
PMC10674526
2.9. Statistical Analysis
trunk FM, FM
In the primary analysis, changes in outcome variables were evaluated using two-way mixed ANOVA with repeated measures, with group specified as a between-subjects factor and time specified as a within-subjects factor. Normality was evaluated by inspection of quantile–quantile plots, data were checked for extreme outlier...
PMC10674526
Sample Size Determination
A priori, a sample size of 50 was selected, with a goal of 20 per treatment group (PRO, PRO + FB) and 10 in the CON group. This sample size was informed by a power analysis using the partial eta-squared effect size for the BFP group × time interaction reported in a previous trial examining the effects of chronic consum...
PMC10674526
3. Results
PMC10674526
3.1. Participant Characteristics
IRREGULAR MENSTRUAL CYCLE
Of the 52 total participants (26 males and 26 females), 28 (54%) reported being concurrently trained (i.e., engaging regularly in both endurance, resistance, and/or concurrent training sessions); 20 (38%) were exclusively resistance-trained; and 4 (8%) were exclusively endurance-trained. Across the entire sample, 37 pa...
PMC10674526
3.2. Participant Compliance
Compliance with completion of daily online surveys was 96.8 ± 7.2%, 91.9 ± 11.0%, and 93.5 ± 13.4% in CON, PRO, and PRO + FB, respectively. From the completed surveys, PRO participants reported following their normal nutritional habits 99.4% of the time, following their normal exercise habits 98.4% of the time, consumi...
PMC10674526
3.3. Energy Intake, Expenditure, and Caffeine Consumption
±
Across the entire sample, at baseline, participants reported consuming 2191 ± 870 kcal, 120 ± 62 g protein, 90 ± 48 g fat, and 227 ± 86 g carbohydrate per day as assessed via the ASA24. At the second visit, participants reported consuming 1997 ± 741 kcal, 128 ± 57 g protein, 81 ± 39 g fat, and 192 ± 84 g carbohydrate. ...
PMC10674526
3.4. Body Composition
No significant group, time, or group × time effects were observed for body composition variables (
PMC10674526
3.5. Body Mass and Anthropometrics
No significant group, time, or group × time effects were observed for body mass, waist circumference, and waist-to-hip ratio (
PMC10674526
3.6. Metabolism and Hemodynamics
A statistically significant group × time interaction was observed for REE values (No significant group, time, or group × time effects were observed for RQ, heart rate, or blood pressure (
PMC10674526
3.7. Sleep Quality, Eating Behavior, and Mood
At baseline, the entire sample of participants demonstrated a sleep quality score of 4.9 ± 2.2 points (range: 1–10), closely centered around the diagnostic cutoff for “poor” sleep of 5 points or greater [
PMC10674526
3.8. Side Effects
ADVERSE EVENTS
No serious adverse events were reported. From daily surveys asking “Are you experiencing any side effects you believe could be related to the dietary supplements you are consuming?”, participants in the PRO condition reported suppressed appetite (
PMC10674526
3.9. Blinding Efficacy
At the end of the study, 9 out of 19 (47%) participants in PRO + FB were able to successfully guess that they had been provided with the active supplement (9 guessed the active supplement, 6 the placebo, and 4 answered, “I don’t know”). Overall, 13 out of 20 (65%) participants in PRO were able to successfully guess tha...
PMC10674526
4. Discussion
PMC10674526
4.1. Body Composition, Body Mass, and Anthropometrics
satiety, weight loss
In the current investigation, four weeks of daily thermogenic supplementation and whey protein ingestion (PRO + FB) did not result in appreciable changes in body mass, body composition, or other anthropometric variables when compared to no supplementation (CON) or to whey protein ingestion alone (PRO). It is important ...
PMC10674526
4.2. Resting Energy Expenditure and Hemodynamic Variables
weight loss
A statistically significant group × time interaction was observed for REE, with values decreasing by approximately 95 ± 107 kcal/day in CON but more closely maintained at −52 ± 128 kcal/day in PRO and +16 ± 133 kcal/day in PRO + FB. A significant group effect was also observed in the one-way ANOVA on REE change values,...
PMC10674526
4.3. Subjective Variables
eating behaviors, mood disturbance, fullness, ’ appetite
No group × time effects were observed for changes in food and eating behaviors, sleep quality, or mood disturbance as assessed via our series of validated questionnaires. Across the entire sample, mood disturbance was reduced (−1.5; Notably, use of the thermogenic supplement did not appear to result in decreased sleep ...
PMC10674526
4.4. Study Strengths and Limitations
The present investigation meaningfully contributes to the current literature in several ways. It includes a mixed-sex cohort of both resistance- and endurance-trained individuals, which is reflective of the populations typically consuming thermogenic supplements [One limitation of the current study is that a group cons...
PMC10674526
4.5. Directions for Future Research
Given the relatively ad libitum nature of the intervention which required no conscious efforts to restrict energy intake or increase physical activity, it is worth considering that these gradual changes may amount to clinically meaningful effects over a longer study duration. Longer-term investigations in active and tr...
PMC10674526
Supplementary Materials
The following supporting information can be downloaded at: Click here for additional data file.
PMC10674526
Author Contributions
Conceptualization, M.R.S., C.R., L.W.T. and G.M.T.; Data curation, M.R.S., C.R., S.J.W., E.T., B.D., A.B., C.L. and J.R.; Formal analysis, M.R.S. and G.M.T.; Funding acquisition, L.W.T. and G.M.T.; Investigation, M.R.S., C.R., S.J.W., E.T., B.D., A.B., C.L. and J.R.; Methodology, M.R.S., C.R. and G.M.T.; Project admini...
PMC10674526
Institutional Review Board Statement
The study was conducted according to the guidelines of the Declaration of Helsinki and approved by the Institutional Review Board of Texas Tech University (protocol code 2022-933, date of approval: 2 November 2022).
PMC10674526
Informed Consent Statement
Informed consent was obtained from all subjects involved in the study.
PMC10674526
Data Availability Statement
Data may be available from the corresponding author upon reasonable request and pending relevant approvals.
PMC10674526
Conflicts of Interest
G.M.T. has received support for his research laboratory through funding or in-kind gifts from nutrition and sports nutrition companies. G.M.T. is the Owner of Tinsley Consulting LLC, which provides paid consulting services to dietary supplement manufacturers. EHP Labs, the sponsor of the present study, is not a past or...
PMC10674526
References
eating behavior
Changes in selected body composition values over the four-week intervention in each group. (Changes in selected body mass and selected anthropometric values over the four-week intervention in each group. (Changes in REE in kcal/day over the four-week intervention in each group.Changes in selected hemodynamic values ove...
PMC10674526
ABSTRACT
PMC10013518
Introduction
Objective structured clinical examination (OSCE) results could be affected by low homogeneity of examiners, non-retrospectiveness of test results, and examiner-cohort effect. In China, many students participate in medical qualification examinations, and this issue is particularly significant. This study aimed to develo...
PMC10013518
Methods
The subjects of this study were clinical students one year after graduation participating in the clinical skills portion of the National Medical Licensing Examination. The participants were from four cities in Jiangsu province. Participants were randomly allocated to on-site and video rating groups to evaluate the rati...
PMC10013518
Results
The reliability of recording equipment and evaluability of video recording were high. Evaluation consistency between experts and examiners was acceptable, and there was no difference in evaluation results (P = 0.61). There was good consistency between video and on-site rating; however, a difference between the two rati...
PMC10013518
Conclusions
Video-based rating could be reliable and offer advantages over on-site rating. The video recording, video-based rating method could provide greater content validity based on its traceability and the ability to view details. Video recording, video-based rating offers a promising mthod for improving the effectiveness and...
PMC10013518
KEYWORDS
PMC10013518
Introduction
The assessment of clinical competence within medical education primarily occurs through written examinations designed to evaluate medical knowledge and performance assessments, such as the objective structured clinical examination (OSCE) to evaluate clinical skills [Studies have demonstrated multiple threats to score r...
PMC10013518
Materials and methods
PMC10013518
Participants
In 2020, 4,714 students participating in the clinical skills portion of the National Medical Licensing Examination from four cities in the Jiangsu province (Nanjing, Suzhou, Nantong, and Xuzhou) were selected. All procedures performed in this study were approved by the ethics committee of Nanjing Medical University and...
PMC10013518
Examination and evaluation methods
abscess
ABSCESS
The clinical skills portion of the National Medical Licensing Examination consisted of three assessment components, with the first station evaluating clinical thinking. The test questions were delivered by a computer. The test duration of the first station was 40 minutes. The second station evaluated physical examinati...
PMC10013518
Video collection
The audio and video recording equipment set consisted of a six-camera surrounding unit, a three-camera independent pickup, and a recording and broadcasting workstation to ensure the critical operations were not obscured. During the exam, the audio and video recording equipment documented the complete process, which was...
PMC10013518
Examiners and scoring
All examiners were experienced clinicians. To improve the homogeneity level of examiners and the efficiency of scoring, the video-based score adopted a classified scoring method, and each examiner only evaluated one type of test. After intensive training, classified training, and test evaluation, the examiners assessed...
PMC10013518
Reliability of equipment and evaluability of video recording
We counted the number of video answer files that should be received and the number of video answer files actually received in the four bases. Equipment reliability was measured as the number of video answer files actually received divided by the number of video answer files receivable. The examiners evaluated the quali...
PMC10013518
Statistical analysis
Measurement data were expressed as the mean ± standard error of the mean (SEM). The non-parametric Wilcoxon rank-sum test evaluated the difference in scores between the two groups with and without video recording in the second and third stations and the difference in video scores between experts and examiners in the se...
PMC10013518
Results
PMC10013518
Reliability and evaluability of video recording
In 2020, the expected number of videos received by candidates from Nanjing, Suzhou, Nantong, and Xuzhou was 1909, 1008, 887, and 910, respectively. The actual number of videos received was 1909, 1008, 887, and 910, respectively, and the missing rate was 0, indicating that the video recording equipment was reliable.Ther...
PMC10013518
Comparison between examiner and expert scoring
A comparative analysis of the differences between the examiner and expert video scoring results revealed that the average score of examiners was 14.45, and the SD was 2.95. The average score of experts was 14.82, and the SD was 2.60. There was no statistical difference between the results of the examiner and experts fo...
PMC10013518
Consistency evaluation of examiner and expert assessment
Based on 9 sample video observations and scoring, the consistency of the average scores of the 12 examiners and expert scores at the second and third stations was evaluated. The results showed that the average scores of the examiners at the two stations and the expert scores were highly consistent. The ICC was 0.94 (Re...
PMC10013518
Comparison analysis of the difference between on-site and video assessment
In clinical skill test, the examiner used a structured score sheet to assess. The results of the second station showed that the average score of video rating was 14.45 (SD = 3.03), which was lower than the on-site rating score (mean = 14.61, SD = 3.06, Scores for on-site and video assessment (Mean (SD)).
PMC10013518
The impact of video recording on student scores
The difference between the on-site rating results of the students with video recording and the on-site scoring results of all students was analyzed. The results showed that there were differences in the scores between the second station and the third station. The scores of all students group were 15.09 (SD = 3.37) and ...
PMC10013518
Discussion
The clinical skill test is an integral part of the physician qualification exam in China. At present, the clinical skill test mainly takes the form of on-site examiner scoring. However, the traditional examiner grading model has certain shortcomings. The test process cannot be reproduced, and test scores cannot be trac...
PMC10013518
Limitations
Although our study showed that video recording and video-based rating had significant strengths in OSCEs, they nonetheless have some limitations. We conducted a statistical analysis to determine whether video recording had an impact on the scores. Although we did not find a significant effect on the scores, further res...
PMC10013518
Future study
Future studies should address the limitations described. The video recording process needs further optimization to ensure the clarity and evaluability of the recording details, providing high-quality video information to the examiner. Studies should explore the perceptions of examiners and participants on the use of vi...
PMC10013518
Conclusions
The traditional scoring model is not sufficiently objective and efficient. This study revealed that the video rating mode could eliminate the examiner’s intervention with the students on-site and improve the objectivity and fairness of examinations. Moreover, the method of categorized scoring could reduce examiners’ wo...
PMC10013518
Acknowledgments
The authors would like to thank all physiotherapy students who participated in this study.
PMC10013518
Disclosure statement
No potential conflict of interest was reported by the authors.
PMC10013518
Data availability statement
The data that support the findings of this study are available from the corresponding author, Hua Huang, upon reasonable request.
PMC10013518
Ethical approval
All data were collected and analyzed solely for evaluation and quality improvement. All procedures performed in this study were approved by the ethics committee of Nanjing Medical University.
PMC10013518
References
PMC10013518
1. Introduction
depression, anxiety
According to animal studies, saffron and its main volatile compound safranal may reduce biological and behavioral signs of acute stress. However, little is known about its impact in humans. This study investigated the acute effect of a saffron extract and safranal on the biological and psychological stress responses in...
PMC10346743
2. Materials and Methods
PMC10346743
2.1. Participants
Twenty healthy men, aged 18–25 years, with a body mass index between 18.5 and 25.0 kg/m
PMC10346743
2.2. Tested Products
Participants were randomly allocated to receive a single dose of 30 mg of a full-spectrum standardized saffron extract (Safr’Inside
PMC10346743
2.3. Study Design and Protocol
This study adopted a randomized, double-blind, placebo-controlled, cross-over (3 arms) design. Each participant went through one screening visit (V0) followed by three testing visits (V1, V2, and V3), during which they consumed either the saffron extract, the safranal or the placebo capsule. Following criteria checks f...
PMC10346743
2.4. Maastricht Acute Stress Test
MAST
The MAST is a validated experimental stress procedure that has been widely used as a safe and reliable measure to activate both acute physiological and subjective measures of the stress response in healthy adults [
PMC10346743
2.5. Salivary Cortisol and Cortisone
Salivary samples were obtained using a Salivette kit (Sarstedt, Nümbrecht, Germany) to measure cortisol and cortisone concentrations. Once collected, samples were stored at 4 °C before being analyzed. Salivary cortisol and cortisone were measured by LC-MS/MS (Prominence liquid chromatography system; Shimadzu, Nakagyo, ...
PMC10346743
2.6. Statistical Analyses
anxiety
As no clinical study has investigated the effect of saffron extract in response to an acute laboratory stressor on salivary cortisol response, the sample size calculation was based on the effect size of a lemon balm food supplement on salivary cortisol in healthy adults 1 h post-treatment [Demographic and lifestyle cha...
PMC10346743
3. Results
PMC10346743
3.1. Inclusion and Population Characteristics
The study population involved 19 healthy males with a mean age of 22.4 ± 2.0 years. All participants reported being healthy, not consuming any food supplements, medications, or illicit drugs that would interfere with the test product. All baseline values were within the physiological range (No statistically significant...
PMC10346743
3.2. Validation of the MAST
anxiety
The analyses of the VAS revealed that the participants, irrespective of treatments, experienced a peak of subjective anxiety and stress 15 min after the MAST (
PMC10346743
3.3. Self-Perceived Anxiety and Stress
anxiety
With regard to anxiety VAS, there was a significant difference in VAS anxiety response (AUCWith regard to stress VAS, there was a significant difference in VAS stress response (AUC
PMC10346743
3.4. Salivary Cortisol and Cortisone Concentration
With regard to salivary cortisol, there was a significant treatment effect regarding the time to reach maximal salivary cortisol concentration (T
PMC10346743
4. Discussion
depression, anxiety
The current study is the first double-blind, randomized, crossover study that explored the effect of a single dose of a proprietary saffron extract (Safr’InsideIrrespective of the treatment, the participants found the MAST to indeed be stressful, as suggested by the increase in perceived anxiety and stress 15 min after...
PMC10346743
5. Conclusions
This study reports that a single dose of Safr’Inside
PMC10346743
Supplementary Materials
The following supporting information can be downloaded at: Click here for additional data file.
PMC10346743
Author Contributions
Conceptualization, P.P. and J.B.; methodology, P.P., J.B. and J.-B.C.; formal analysis, C.P.; investigation, P.P.; data curation, J.B. and J.-B.C.; project administration, P.P.; writing—original draft preparation, C.P., N.C. and L.C.; writing—review and editing, C.P.; visualization, C.P. and L.P.; supervision, P.P.; fu...
PMC10346743
Institutional Review Board Statement
The study was conducted in accordance with the Declaration of Helsinki and the French Public Health Code. The study protocol was approved by the Ethics committee for people’s protection (CPP EST IV, 28/09/2019; ID-RCB: 2018-A02178-47). The trial was registered at ClinicalTrials.gov (NCT04523870).
PMC10346743
Informed Consent Statement
Informed consent was obtained from all subjects involved in the study.
PMC10346743
Data Availability Statement
The data presented in this study are available on request from the corresponding author.
PMC10346743
Conflicts of Interest
C.P, L.P. and D.G. work for Activ’Inside and provided the tested products.
PMC10346743
References
anxiety
Consolidated Standards of Reporting Trials (CONSORT) flowchart diagram. S1: saffron extract; S2: safranal; P: placebo.Effect of the Maastricht acute stress test (MAST) on self-perceived anxiety (Mean and SEM of self-perceived anxiety (Mean and SEM of Tmax for salivary cortisol and cortisone by treatment. Placebo (in bl...
PMC10346743