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Despite comprising half the population, females are often left out of muscle research due to the impact of changing hormones during the menstrual cycle and when using oral contraceptives. This makes it hard to perform costly and invasive studies involving tracers to study muscle protein metabolism. Consequently, we lack a clear understanding of how these hormonal changes affect muscle growth.
There is a need for less invasive methods to study how sex hormones and oral contraceptives influence muscle protein metabolism. Ex vivo models, where serum from participants is applied to mouse muscle cell cultures, mimic the conditions of human muscle cells and can provide initial insights.
The aim of the study is to develop a non-invasive model using serum from both oral contraceptive users and non-users at various stages of their cycles, to understand if different cycle or pill stages affect how muscles process proteins.
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| Label | Type | Description | Intervention Names |
|---|---|---|---|
| Mid-Follicular Phase | Experimental | 7-11 days after onset of menses. |
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| Mid-Luteal Phase | Experimental | 5-9 days after ovulation (as confirmed with ovulation test kits). |
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| Active pill phase | Experimental | 10-20 days after starting new pill cycle. |
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| Withdrawal phase | Experimental | 48hrs after last pill (during placebo pill phase). |
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| Name | Type | Description | Arm Group Labels | Other Names |
|---|---|---|---|---|
| Protein tracer drink | Behavioral | Subjects will be fed a mixed-macronutrient beverage at rest (0.75g/kg lean body mass of carbohydrates; 0.125g/kg lean body mass of protein). Amino acid composition of the protein will be modelled off the composition of egg. Leucine content will be enriched to 5% with [13 Carbon(13C)]-leucine. |
| Measure | Description | Time Frame |
|---|---|---|
| Protein Synthesis (Murine Cell-Based Experiments, ex-vivo experiments) | Investigators will use human serum obtained from fasted and fed timepoints (-15, 20, 40 and 60 minutes following beverage consumption) to condition cell culture media (20% volume). To determine the effects of using fasted and/or fed 'human-conditioned' culture media on cell protein synthesis, puromycin incorporation (measure of protein synthesis) will be measured via western blot and expressed relative to a no-serum control. A two-way repeated measures ANOVA will be used to analyze outcomes with cycle stage and group (OC vs non-OC) used as factors | 60 minutes |
| Measure | Description | Time Frame |
|---|---|---|
| Whole-body protein synthesis | Investigators will measure the enrichment of [13 Carbon CO2 (13CO2)] in the breath by isotope ratio mass spectrometry (IRMS) in atom percent excess (APE). The measurement of carbon dioxide production (VCO2) and stable isotope tracer enrichment in the breath allows for the assessment of the rate at which amino acids are used for energy (i.e., oxidized), rather than for protein synthesis (i.e., retained in the body) by calculating the fraction of expired CO2 that contains 13C. Leucine retention (umol/kg) will then be calculated from the difference between the known amount of leucine provided (ingested) and leucine oxidation (as determined from 13CO2 breath enrichment). |
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Inclusion Criteria:
Exclusion Criteria:
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| Name | Role | Phone | Extension | |
|---|---|---|---|---|
| Daniel R Moore, PhD | Contact | 4169464088 | dr.moore@utoronto.ca | |
| Ines Kortebi, PhD Student | Contact | ines.kortebi@mail.utoronto.ca |
| Name | Affiliation | Role |
|---|---|---|
| Ines Kortebi, PhD Student | University of Toronto | Study Director |
| Cassidy Tinline-Goodfellow, PhD (C) | University of Toronto | Study Director |
| Jonathan Aguilera, PhD Student |
| Facility | Status | City | State | ZIP | Country | Contacts |
|---|---|---|---|---|---|---|
| Goldring Centre for High Performance Sport at the University of Toronto | Recruiting | Toronto | Ontario | M5S2C9 | Canada |
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This study uses a 4-way counterbalanced crossover design. A total 10 young women (Ages: 18-35 years old; 5 per OC and non-OC group) will be recruited and randomized to begin in either the mid-follicular (7-11 days after the onset of menses) or mid-luteal phase (5-9 days after ovulation) if part of non-OC group; and active pill (10-20 days within starting their new pill cycle) or withdrawal phase (at least 48hrs after their last pill) if part of the OC group. Participants of each group will take part in two metabolic trials, one in each phase of their menstrual cycle or pill cycle.
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| 6 hours |
| Urinary Measures (Muscle Protein Breakdown) | Investigators will measure urinary 3-methylhistidine (3MH) as an indirect marker of muscle protein breakdown over the course of the trial (6 hours) through pooled urine collection vs baseline urine. | 6 hours |
| University of Toronto |
| Study Director |