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Descriptive statistics
The means and standard deviations for compassion, relevance, and interest in each video are presented in Table  Relevance(Range: 13.33–100)
PMC10061748
Engagement in videos among women
All descriptive statistics for ANOVAs and ANCOVA analyses for women examining the differences in engagement by video condition are displayed in Table  Note:
PMC10061748
Engagement in videos among men
As measures of pre-test muscularity, weight, and shape dissatisfaction were unrelated to interest in women, a univariate ANOVA was conducted to examine the differences between video conditions on interest. There was no significant effect of video condition on interest, As measures of pre-test weight satisfaction, shape...
PMC10061748
Discussion
SE, cognitive and affective, eating disorders, cognitive, affective, and behavioural
As measures of pre-test muscularity, weight, and shape dissatisfaction were unrelated to interest in men, a univariate ANOVA was conducted to examine the differences between video conditions on interest. There was no significant effect of video condition on interest, As measures of pre-test weight satisfaction, shape s...
PMC10061748
Strengths and Limitations
eating disorders
A strength of this study was its generalisability; a mixed-age, community sample of men and women was used, which is not common in evaluating social marketing or anti-stigma campaigns for body image and eating disorders [Further limitations were that engagement was examined at one point in time, which precluded examina...
PMC10061748
Acknowledgements
Thank you to The Mannequin Project at Big Picture Storytelling for video production and permission to use the videos in this research.
PMC10061748
Authors’ contributions
Jo Doley: conceptualisation, formal analysis, data curation, writing – original draft preparation, writing – review and editing. Siân McLean: conceptualisation, data curation, supervision, methodology, resources, investigation, writing – original draft preparation, writing – review and editing, project administration, ...
PMC10061748
Funding
This research was partially supported by funding from Australian Rotary Health to the second author.
PMC10061748
Data Availability
The datasets generated and/or analysed during the current study are not publicly available as this was not a condition outlined to our participants, but are available from the corresponding author on reasonable request.
PMC10061748
Declarations
PMC10061748
Ethics approval and consent to participate
This study was approved by the La Trobe University Human Research Ethics Committee, approval number HEC15-116. All methods were carried out in accordance with the National Statement on Ethical Conduct in Human Research, the Australian Code for the Responsible Conduct of Research and Commonwealth and state laws. All par...
PMC10061748
Consent for publication
Not applicable.
PMC10061748
Competing interests
The authors have no competing interests to declare.
PMC10061748
References
PMC10061748
Subject terms
hypoxia, fatigue, impairments in exercise performanceMoreover
HYPOXIA
Ten male cyclists were randomized into four experimental conditions in this randomized, cross-over, double-blind, and sham-controlled study to test the combined effect of acute dark chocolate (DC) ingestion and anodal concurrent dual-site transcranial direct current stimulation (a-tDCS) targeting M1 and left DLPFC on c...
PMC10542360
Results
PMC10542360
Effect of the demanding cognitive task
fatigue
Performing the incongruent only Stroop task for 30 min did not induce a state of mental fatigue as we found no significant main effect of time Reaction time changes in the four experimental conditions. Comparison of choice reaction time performance (
PMC10542360
Effect of DC and a-tDCS
Our results showed a significant condition x time interaction
PMC10542360
Effect of DC and a-tDCS
The concomitant use of DC and a-tDCSEndurance performance and physiological responses in the four experimental conditions. Time to exhaustion test (
PMC10542360
Effect of DC and a-tDCS
DC and/or a-tDCS
PMC10542360
Effect of DC and a-tDCS
RPE
DC and/or a-tDCSPsychophysiological responses to exercise in the four experimental conditions. Ratings of perceived exertion (RPE;
PMC10542360
Discussion
hypoxia, cognitive improvements
HYPOXIA, VASODILATION
The main results of the present study were that (1) ‘DC + a-tDCS’ resulted in longer TTE in hypoxia than ‘WC + a-tDCS’ and ‘WC + sh-tDCS’ conditions, (2) with greater EMG activity of the VM muscle, and (3) improved cognitive performance during and after TTE. These results suggest the possibility of a synergistic effect...
PMC10542360
Methods
PMC10542360
Participants
Ten endurance-trained male cyclists voluntarily participated in this randomized, sham-controlled, and double-blind trial. The general characteristics of the participants are presented in Table General characteristics of the participants (n = 10).
PMC10542360
General experimental procedure
hypoxia
HYPOXIA
The participants visited the laboratory on six different occasions at one-week intervals. In the first session, participants underwent an anthropometric assessment and were familiarized with the whole experimental procedure. An individual not involved in the research team randomized the order of four experimental condi...
PMC10542360
Maximal incremental test
hypoxia
HYPOXIC, HYPOXIA
In the second session, after 30 min being under hypoxia at rest, participants underwent the maximal incremental test (Astrand Test for Men) on a cycle ergometer (Cyclus 2, RBM Elektronik-Automation GmbH, Leipzig, Germany) to measure PPO under the hypoxic condition. The test was started with 100 watts and a cadence of 6...
PMC10542360
Dark chocolate supplementation
In the DC conditions (DC), 2 h before the experimental sessions participants consumed ~ 467 kcal of a typical commercial 70% cocoa product (Nestlé Noir 70%) containing the ingredients cocoa liquor, sugar, cocoa butter, milk fat, lecithin and vanilla
PMC10542360
Cognitive function—choice reaction time
hypoxia
HYPOXIA
The Visual Choice Reaction Time Apparatus (Model 63035A, Lafayette Instrument Company, Indiana, USA) with a four-choice compatible stimulus–response paradigm was used to measure cognitive performance. Three visual stimuli (lights turning on) were manually given to the participants, and they were instructed to respond a...
PMC10542360
MIVC
At the beginning of each session, participants performed 3–5 s knee extension MIVC three times with a 150-s rest in between on a custom-made chair with knee and hip fixed at 90° as recommended for VL, VM, and RF muscles MIVC test
PMC10542360
EMG
The surface EMG signals were collected strictly according to the recommended standards
PMC10542360
Hypoxic exposure
HYPOXIC
The hypoxic condition (O
PMC10542360
Prolonged cognitive effort
hypoxia
HYPOXIA
Participants performed a modified computerized version of CWST (100% incongruent) for 30 min under hypoxia. The incongruent CWST consisted of four-color words (yellow, blue, green, and red) in different font inks (yellow, blue, green, and red) that were randomly displayed on a computer screen until the participant ente...
PMC10542360
Transcranial direct current electrical stimulation (tDCS)
CORTEX
Two-channel battery-driven stimulators (NeuroStim 2, Medina Tebgostar, Tehran, Iran) were used to apply tDCS over the target brain areas during the 3rd to 6th experimental sessions (two devices were used). Four carbon electrodes covered by saline-soaked surface sponges (NaCl 140 mmol dissolved in Milli-Q water) were us...
PMC10542360
tDCS modeling
The brain current flow during tDCS was calculated using a finite element model (FEM) following the standard pipeline in SimNIBS 4.0.0
PMC10542360
tDCS-induced sensations and blinding assessment
Participants completed a questionnaire provided by Fertonani et al.
PMC10542360
Whole-body exhausting endurance task
hypoxia
HYPOXIA
Within the 3rd to 6th sessions, after 30 min performing CWST, followed by 20 min of tDCS, participants performed the TTE on a cycle ergometer (Cyclus 2, RBM Elektronik-Automation GmbH, Leipzig, Germany), all under hypoxia. Before TTE, participants warmed up cycling for 5 min at 45% PPO. After warming up, TTE started wi...
PMC10542360
HR and SpO
During the whole experimental session, HR was continuously monitored by the use of a chest strap (M430, Polar, Finland) connected to the cycle ergometer. SpO
PMC10542360
Ratings of perceived exertion
RPE, hypoxia
HYPOXIA
The 0–100 Borg scale was used to measure RPE. Participants received the instruction to report their RPE at the end of each stage and the point of exhaustion in the incremental test, and every 3 min and upon reaching the point of exhaustion in the TTE in hypoxia
PMC10542360
Affective responses and felt arousal
bipolar
The affective responses were reported using the Feeling Scale (FS), which is a bipolar scale comprising eleven items ranging from  − 5 (very bad) to + 5 (very good), validated by Hardy and Rejeski
PMC10542360
Statistical analyses
The normal distribution of each data set was evaluated by the Shapiro–Wilk normality test. Values are presented as means and standard deviation (SD) or median and interquartile range (IQR) as stated. The Friedman test was used to compare tDCS-induced sensations, end-of-study guess rate accuracy, and active stimulation ...
PMC10542360
Acknowledgements
The authors would like to thank Iman Talebi-Rasa and Matin Etemadi for their help in data acquisition. The authors also express their sincere gratitude to the participants for their commitment to the experimental procedure. This research was conducted in the Exercise Metabolism and Performance Lab (EMPL), Faculty of Sp...
PMC10542360
Author contributions
P.B., V.T., E.A., and D.M. conceptualized and designed the study. P.B., V.T., and E.A. conducted the experiments. P.B., V.T., E.A., and D.M. participated in the formal analysis. P.B. wrote the original draft of the manuscript. V.T., E.A., and D.M. reviewed and edited the manuscript. All authors read and approved the fi...
PMC10542360
Funding
This research was conducted in the Exercise Metabolism and Performance Lab (EMPL), Faculty of Sport Sciences, Razi University, Kermanshah, Iran.
PMC10542360
Data availability
The datasets used and/or analyzed during the current study are available from the corresponding author upon reasonable request.
PMC10542360
Competing interests
The authors declare no competing interests.
PMC10542360
References
PMC10542360
Keywords
CONTRACTION, CONTRACTIONS
Communicated by Francesco Lacquaniti.This study used transcranial magnetic stimulation (TMS) to determine if muscarinic receptor blockade affects muscle responses during voluntary contractions. Motor evoked potentials (MEPs) were recorded from biceps brachii in 10 subjects (age: 23 ± 2) during 10%, 25%, 50%, 75%, and 1...
PMC10224869
Introduction
gain of motor circuits, voluntary muscle contraction, muscle fatigue, voluntary muscle contractions
CONTRACTION, CORTEX
Four primary neuromodulator systems exist in the central nervous system (CNS): dopaminergic, adrenergic, serotonergic, and cholinergic. While there is a good understanding of how the first three systems regulate the gain of motor circuits in the CNS, our understanding of how the cholinergic system regulates the excitab...
PMC10224869
Methods
PMC10224869
Participants
Ten healthy individuals (mean age 23 ± 2) were involved in the study. Each participant attended two sessions spaced 10 days apart. Participants were screened using a modified medical history questionnaire that identified contraindications to promethazine, TMS, percutaneous electrical stimulation, and the exercise tasks...
PMC10224869
Compliance with ethical standards
Written and informed consent was obtained before the commencement of testing. This experiment was approved by the Griffith University Human Research Ethics Committee and conformed to the standards set by the Declaration of Helsinki.
PMC10224869
Experiment design and drug administration
This study was a human, double-blinded, placebo-controlled, crossover design. Participants were administered either a placebo or 25 mg promethazine capsule. Promethazine was chosen for its high potency and antagonistic affects upon mAChRs (Kubo et al.
PMC10224869
Force and EMG recording
Participants were seated with their elbow flexed and held at 90° in a custom designed force transducer (Fig. Participants were seated with their arm attached to a custom designed force transducer. Biceps brachii EMG was recorded from the right limb following the administration of promethazine or a placebo (
PMC10224869
Motor cortical stimulation
CORTEX
A circular coil with a 90 mm outside diameter was positioned over the vertex of the head and delivered single-pulse stimulations to the motor cortex via a MagStim 200
PMC10224869
Active motor threshold (AMT)
CONTRACTION
Each participant’s motor threshold was determined during light contraction of the biceps brachii. To ensure consistency between participants and between sessions, a horizontal cursor of 0.01% peak-to-peak Mmax was displayed on a monitor, and participants were required to track the cursor with their rectified biceps bra...
PMC10224869
TMS stimulus–response curves
Muscle responses were obtained from the lightly contracted biceps brachii from 90% AMT to 190% AMT in 10% AMT increments in a semi-randomised protocol. Six stimulations were delivered for each of the 11 stimulator intensities, where a 20 s rest was provided between each stimulation. One participant did not reach 190% A...
PMC10224869
Contraction protocol 1: unfatigued muscle
torques
CONTRACTION, CONTRACTIONS
Participants performed 5 maximal effort contractions, where the trial that produced the largest torque was deemed the participants maximal voluntary contraction (MVC). Cortical and brachial plexus stimulation was delivered during initial baseline contractions, with a minimum of 2 min rest occurring between each effort....
PMC10224869
Contraction protocol 2: fatigued muscle
fatigue
CONTRACTION, CONTRACTIONS
After the unfatigued protocol was completed, a second protocol was performed that involved a fatigue-inducing sustained maximal contraction. Each participant held a maximal contraction until their force had reduced to 60% of their initial unfatigued MVC. This ensured that each trial, and each participant, had the same ...
PMC10224869
Data analysis
EVENT, CONTRACTION
Data were analysed using built-in functions in Spike2 v7.02a software (Cambridge Electronic Design). MEP area and Mmax area were calculated from biceps brachii EMG following cortical and brachial plexus stimulations, respectively. MEP area and Mmax area was calculated from the first deflection from baseline created by ...
PMC10224869
Statistical analysis
All statistical analyses were performed in R, using RStudio (version 2022.02.0 + 443 "Prairie Trillium" release, Boston, MA) where significance level of Input–output curve for MEP area (
PMC10224869
Results
PMC10224869
Active motor threshold and TMS stimulus–response
Promethazine did not affect AMT as there were no drug-related differences in stimulator output when MEPs were first detected in the lightly contracting muscle (
PMC10224869
Maximal voluntary contraction torque
Promethazine did not affect maximal elbow flexion torque as there were no drug-related differences in MVC (
PMC10224869
MEP and silent period duration during brief voluntary contractions.
CONTRACTIONS
There was no main effect of drug on MEP area during brief unfatigued voluntary contractions (Protocol 1 MEP area (There was a main effect of drug identified for the TMS-evoked SP (
PMC10224869
MEP and silent period duration following fatiguing sustained contractions
CONTRACTIONS
There was no main effect of drug on MEP area following the maximal effort sustained contractions (Protocol 2 MEP area (Unlike the unfatigued contractions, there was no main effect of the drug detected for TMS-evoked SP duration following the sustained voluntary contractions (
PMC10224869
Discussion
voluntary muscle contractions, fatigue
CONTRACTION, CONTRACTIONS
The present study assessed the effects of a potent antimuscarinic drug on MEP area and TMS-evoked SP during voluntary contractions. To achieve this, a contraction protocol was used to assess cholinergic effects during brief contractions of 10%, 25%, 50%, 75%, and 100% MVC. This was followed by a second protocol that as...
PMC10224869
Silent period is affected by antimuscarinic activity during unfatigued voluntary contractions
voluntary muscle contractions
CONTRACTION, CONTRACTIONS
The current study revealed that SP duration shortened during voluntary muscle contractions, where the duration of TMS-evoked SP progressively reduced with an increase in contraction intensity. The relationship between SP and contraction intensity has been reported on a number of occasions, where many studies report tha...
PMC10224869
Antimuscarinic activity did not affect MEP area during unfatigued voluntary contractions
CONTRACTION
A hypothesis of the current study was that mAChRs would modulate MEPs. However, there were no antimuscarinic effects detected for MEP area measured during a full range of voluntary contraction intensities. While the TMS-evoked SP involved inhibitory processes that are mediated by GABA
PMC10224869
The cholinergic system did not modulate MEPs or SPs under conditions of fatigue
failure of the muscle, fatigue
CONTRACTIONS
Fatigue-inducing contractions cause a reduction in the maximal force generating capacity of the muscle, where the decline in force may be due to an inability of the nervous system to activate the muscle, or a failure of the muscle itself to contract. The current study normalised the amount of fatigue that was induced f...
PMC10224869
Considerations
Although we have attributed our findings for the motor system to anticholinergic effects, we must acknowledge that the medication used in this study also has strong antihistaminergic effects. We have previously used single choice, and multiple choice, reaction time paradigms to demonstrate that CNS acting antihistamine...
PMC10224869
Funding
Open Access funding enabled and organized by CAUL and its Member Institutions. The authors did not receive support from any organization for the submitted work. No funding was received to assist with the preparation of this manuscript. No funding was received for conducting this study. No funds, grants, or other suppor...
PMC10224869
Data availability
The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.
PMC10224869
Declarations
PMC10224869
Ethical approval
This experiment was approved by the Griffith University Human Research Ethics Committee and conformed to the standards set by the Declaration of Helsinki.
PMC10224869
References
PMC10224869
Background
advanced-stage solid, tumors
GSK3368715, a first-in-class, reversible inhibitor of type I protein methyltransferases (PRMTs) demonstrated anticancer activity in preclinical studies. This Phase 1 study (NCT03666988) evaluated safety, pharmacokinetics, pharmacodynamics, and preliminary efficacy of GSK3368715 in adults with advanced-stage solid tumor...
PMC10338470
Methods
TEEs
THROMBOEMBOLIC EVENT
In part 1, escalating doses of oral once-daily GSK3368715 (50, 100, and 200 mg) were evaluated. Enrollment was paused at 200 mg following a higher-than-expected incidence of thromboembolic events (TEEs) among the first 19 participants, resuming under a protocol amendment starting at 100 mg. Part 2 (to evaluate prelimin...
PMC10338470
Results
toxicities, tumor
PULMONARY EMBOLISM, TUMOR, DISEASE, BEST, EVENTS
Dose-limiting toxicities were reported in 3/12 (25%) patients at 200 mg. Nine of 31 (29%) patients across dose groups experienced 12 TEEs (8 grade 3 events and 1 grade 5 pulmonary embolism). Best response achieved was stable disease, occurring in 9/31 (29%) patients. Following single and repeat dosing, GSK3368715 maxim...
PMC10338470
Conclusion
Based on higher-than-expected incidence of TEEs, limited target engagement at lower doses, and lack of observed clinical efficacy, a risk/benefit analysis led to early study termination.
PMC10338470
Trial registration number
NCT03666988.
PMC10338470
Subject terms
PMC10338470
Background
tumor
TUMOR
Arginine methylation is an important posttranslational modification of proteins involved in diverse cellular processes such as gene regulation, ribonucleic acid (RNA) processing, mRNA splicing, deoxyribonucleic acid (DNA) repair, and signal transduction [GSK3368715 is a potent, reversible, S-adenosylmethionine (SAM)-un...
PMC10338470
Methods
PMC10338470
Study design
This was a Phase 1, open-label study consisting of a dose escalation, followed by a planned dose-expansion cohort of oral administration of GSK3368715 conducted between October 26, 2018 and March 4, 2021 (Fig. 
PMC10338470
Planned study design.
thrombosis, tumor, DLTs, toxicity, toxicities, TEEs, chemotherapy-associated venous thromboembolism, PK/PD, DLBCL, PD
THROMBOSIS, DISEASE PROGRESSION, TUMOR, THROMBOEMBOLIC EVENT, ADVERSE EVENTS, EVALUABLE, REGRESSION, DIFFUSE LARGE B-CELL LYMPHOMA
DLBCL diffuse large B-cell lymphoma, PD pharmacodynamic, PK pharmacokinetic, po orally, qd once daily, RP2D recommended Phase 2 dose. Part 1 of this study included a dose-escalation cohort to assess the incidence of dose-limiting toxicities (DLTs) and adverse events, and a PK/PD cohort to characterize the PK/PD profile...
PMC10338470
Study population
venous thromboembolism
Full inclusion/exclusion criteria for both parts 1 and 2 are included in Supplemental Table As mentioned above, following the study amendment, patients at high risk of venous thromboembolism as defined by either Khorana Score of ≥3, or prior medical history of venous thromboembolism, were ineligible.
PMC10338470
Study assessments
tumor, Tumor, cancer, PK/PD, partial loss, PD
TUMOR, TUMOR, ADVERSE EVENT, CANCER, BLOOD
Adverse events were coded using the standard MedDRA groupings and graded according to NCI-CTCAE version 5.0. Clinical chemistry, urinalysis, and coagulation tests were performed predose on days 1, 8, 15, 22, and weekly thereafter. Tumor imaging occurred every 8 weeks until week 33, and then every 16 weeks thereafter.In...
PMC10338470
Statistical analyses
toxicity, tumor, DLTs, PK/PD
ADVERSE EVENT, TUMOR, BEST
The Neuenschwander Continual Reassessment Method dose-escalation design has been previously described. Briefly, the dose level with the highest posterior probability of having a DLT rate within the target toxicity range (≥16% and <33%) was recommended for the next cohort. Additionally, following the protocol amendment,...
PMC10338470
Results
PMC10338470
Pharmacokinetics
Following single and repeated oral administration, GSK3368715 was rapidly absorbed, with maximum plasma concentration (Two metabolites (GSK3963583, GSK3983164) rapidly formed with time to
PMC10338470
Pharmacodynamics
SE
Reduction of levels of ADMA-R225-hnRNP-A1 in PBMCs was time dependent with a mean (SE) reduction of 54.7% (6.92%) in the GSK3368715 200-mg dose group on day 15. A mean (SE) day 15 reduction of 43.1% (5.81%) was observed at 100 mg (Fig. 
PMC10338470
Discussion
tumor, cancers, TEEs, cancer, pleomorphic adenoma, tumors, MTAP loss
TUMOR, CANCERS, CANCER, DISEASE, ADVANCED CANCER, PERIVASCULAR EPITHELIOID CELL TUMOR, DELETION, PLEOMORPHIC ADENOMA, TUMOR GROWTH, TUMORS, DISEASE CHARACTERISTIC
GSK3368715 is a first-in-class type I PRMT inhibitor that exhibited strong anticancer activity in preclinical studies. Despite this encouraging finding, and early evidence in the current study of target engagement in peripheral blood at doses of 200 mg, the study was paused due to concern over a higher-than-expected ra...
PMC10338470
Conclusions
TEEs, tumor, cancer
CANCER, TUMOR
Despite promising preclinical results and observed peripheral target engagement at higher doses, the incidence of TEEs, variable target engagement at the tumor level, and observed limited clinical efficacy led to early termination of this trial. It is not known whether the lack of clinical efficacy and elevated risk of...
PMC10338470
Supplementary information
The online version contains supplementary material available at 10.1038/s41416-023-02276-0.
PMC10338470
Acknowledgements
HORNER, BRUCE
All listed authors meet the criteria for authorship set forth by the International Committee for Medical Journal Editors. Editorial support (Allyson Lehrman, DPM, assembling tables and figures, collating author comments, copyediting, fact checking, and referencing) and graphic services were provided by AOIC, LLC and we...
PMC10338470
Author contributions
ABE-K, TN, IG-L, EC, JR, BT, PJO’D, AC, and ARAR contributed to the conception or design of the study. ABE-K, EC, JR, BT, PJO’D, AC, and ARAR contributed to the acquisition of the data. JC, TN, TC, NR, CR, PN, AT, IG-L, and AC data analysis or interpretation. ABE-K, JC, TN, TC, NR, CR, PN, AT, IG-L, EC, JR, BT, PJO’D, ...
PMC10338470
Funding
Authors who are or were employees of GSK contributed to the design of the study, analysis of the data, and in the decision to publish. Funding for this study (NCT03666988 available from
PMC10338470
Data availability
Within 6 months of this publication, anonymized individual participant data, the annotated case report form, protocol, reporting and analysis plan, dataset specifications, raw dataset, analysis-ready dataset, and clinical study report will be available for research proposals approved by an independent review committee....
PMC10338470
Competing interests
Cancer, Oncology/Cancer
ONCOLOGY, CANCER, FOUNDER, EMD
ABE-K reports honoraria from ABL bio, Agenus, AstraZeneca/MedImmune, Bayer, Bristol-Myers Squibb, CytomX Therapeutics, EISAI, EMD Serono, Exelixis, Gilead Sciences, Merck, Pieris Pharmaceuticals, QED Therapeutics, Roche/Genentech, SERVIER, and Tallac Therapeutics; consulting or advisory board roles with BL Bio, Agenus,...
PMC10338470
Ethics approval and consent to participate
Cancer
DEL, CANCER
The study was approved by the ethics committee at every participating institution [Melbourne Health Human Research Ethics Committee Parkville, Australia; University Health Network Research Ethics Board, Toronto, Canada; Comite Etico De Investigacion Clinica con medicamentos (CEIm) CEIC del Hospital Universitari Vall d’...
PMC10338470