Evaluation of a novel alternative protein source to stimulate post-exercise muscle protein synthesis

From p8 of C1 protocol document:

Sufficient dietary protein ingestion on a daily basis is needed to supply the
muscles with amino acids to enable the process of muscle protein synthesis.
However, the production of sufficient amounts of animal-based protein from
conventional sources to meet future global food demands represents a challenge.
Edible insects have been proposed as an alternative source of dietary protein
that can be produced on a viable and more sustainable commercial scale and, as
such, may contribute to ensuring global food security. Many edible insects
represent a rich source of protein, comparable to conventional meat and fish,
and provide EAA in amounts comparable to certain high quality protein sources.
However, there is currently limited data on the functional capacity of
insect-based protein sources.

From p11 of C1 protocol document:

1) To define the characteristics of mealworm protein ingestion on
protein/digestion absorption kinetics and both whole-body and myofibrillar
muscle protein synthesis after a single bout of resistance exercise in young
men and 2) compare the response of mealworm vs. milk protein ingestion on
protein/digestion absorption kinetics and both whole-body and myofibrillar
muscle protein synthesis after a single bout of resistance exercise in young
men.

From p12-13 of C1 protocol document:

2.1 Screening
When volunteers respond to the advertisement, we will contact them by
e-mail/phone and briefly explain the study. We will provide them with the
information brochure and the informed consent (which they will bring during the
screening). To assess whether volunteers are eligible to participate in this
study, we will invite them to the University for screening. Before we start the
screening, we will explain the entire experimental trial and answer any
potential questions. We will then ask them to read, fill out, and sign the
informed consent form. After signing the informed consent form, we will start
the screening by going through the medical questionnaire to assess their
general health, use of medication, and physical activity. Subsequently, we will
assess body composition by performing a dual-energy X-ray absorptiometry (DEXA)
scan and measure body height and body weight. DEXA is a simple and non-invasive
procedure, which will take place at the University. Subjects will be instructed
to lie down on a table and stay motionless for approximately 3 minutes during
which the body scan takes place. Performing the above mentioned tests allow us
to characterize the participants. In case of an unexpected medical finding, it
is our duty to inform the subjects. If a participant does not want to receive
this information, he cannot participate in this study.

Following the DEXA scan and anthropometric measurements, subjects will be
familiarized and tested for strength on the exercise machines. Subjects will be
instructed on proper single legged weight-lifting technique on each exercise
machine (leg-press and leg-extension) and complete a standardized testing
protocol to determine a measurement of maximal strength (1RM) for one leg on
each exercise machine. The testing protocol requires that the subjects complete
sets on each exercise machine increasing in weight until volitional fatigue
occurs, ideally occurring between 3-6 repetitions on the heaviest weight. The
attained strength data will be compared to previously published data and used
to calculate an estimation of 1RM. Following the determination of 1RM, subjects
will be scheduled for their experimental testing day.

2.2 Experimental trial
An outline of the study protocol is shown in Figure 1. At 8.00 am, following an
overnight fast, subjects will arrive at the laboratory by car or public
transportation. Subjects will rest in a supine position and a Teflon catheter
will be inserted into an antecubital vein for intravenous stable isotope
infusion. A second Teflon catheter will be inserted in a heated dorsal hand
vein of the contralateral arm and placed in a hot-box (60°C) for arterialized
blood sampling. Following basal blood collection (8 mL; t= -180 min), the
plasma phenylalanine and tyrosine pool will be primed with a single intravenous
dose of tracer and a continuous tracer infusion will commence. Arterialized
blood samples (8 mL) will then be drawn at t= -180, -120, -60 and 0. Starting
at -60 (min), subjects will perform unilateral resistance exercise. Immediately
after the exercise subjects will return to the resting supine position and
arterialized blood sample will be drawn. Afterwards, a muscle biopsy from both
legs will be collected (t= 0 min). Subjects will then receive their respective
nutritional treatment (t= 0). Arterialized blood samples (8 mL) will be
collected at t= 20, 40, 60, 90, 120, 150, 180, 240 and 300 min during the
post-exercise recovery period. At 120 min, again a muscle biopsy will be taken
from both legs but from a different incision. Finally, at 300 min the last
biopsies will be taken from both legs. In total, six muscle biopsies will be
taken through six separate incisions during the trial. The muscle biopsies
(immediately, 2 h, and 5 h after exercise) will allow us to measure the
temporal response of muscle protein synthesis of mealworm and milk protein
ingested after exercise. The muscle biopsies at 0, 2 and 5 hours post-exercise
will provide us with a better insight in the differences between *early* (0-2
h) and *late* (2-5 h) muscle protein synthesis response. The first blood sample
at t= -180 together with the first biopsy from the rested leg at t= 0 allow us
to calculate resting muscle protein synthesis rates prior to exercise and
protein ingestion.

From p17 of C1 protocol document:

Subjects will perform unilateral resistance exercise and consume either 30 g
mealworm or milk protein. In addition, continuous intravenous tracer infusions
will be applied, with plasma and muscle samples collected. The exercise session
consists of a 5 min unilateral warm-up on a cycle ergometer and 4 unilateral
sets on the leg-press and leg-extension machines. The workload is set at 80% of
the subjects* one-repetition maximum (8*10 repetitions) to stimulate muscle
hypertrophy. Resting periods of 2 min are allowed between sets and 2 min
between exercises.

From p31-32 of C1 protocol document:

The burden and risks associated with participation are medium. Insertion of the
catheters is comparable to a blood draw and could result in a small hematoma.
Muscle biopsies will be taken under local anesthesia by an experienced
physician, but may cause some minor discomfort for maximally up to 24 h after
completion. The discomfort is comparable to muscle soreness or the pain one has
after bumping into a table. We will take 13 blood samples (104 mL total) during
the experimental trial respectively. The total amount of blood we draw is less
than half the amount of a blood donation and will be completely restored in
approximately 1 month. For the experimental trial, participants have to be
fasted, so they are not allowed to eat and drink (except for water) from 22h00
the evening before. Also 3 days prior to the experimental trial participants
should not perform any type of intense physical exercise, and should not
consume alcohol. Thereby, participants will be asked to keep their diet as
constant as possible for 3 days and to fill out a dietary and activity record
for 3 days prior to the experimental trial.
The milk protein used in the beverages is similar to normal milk and also
commercially available as a food product, same as the mealworm protein.
Therefore, the test beverages do not form any health risks. The stable isotope
amino acids tracers applied in this experiment are not radioactive and are
completely safe. The production of the tracers for intravenous administration
will occur in a sterile environment according to GMP guidelines.
There is no risk associated with the DEXA scan. The radiation dose emitted
during a DEXA scan is 0.001 mSv. This is a very low exposure compared to the
total background radiation in the Netherlands, which is ~2.5 mSv/year. For
comparison, the radiation dose during a flight higher than 10 km is 0.005
mSV*h-1. Performing exercise will pose little risk as the possibility for
adverse health events will be evaluated during the screening and subjects will
be closely monitored throughout the exercise session.
There is no direct benefit for the participants, only their contribution to
scientific knowledge on insects as a sustainable alternative source of dietary
protein for human consumption.