Trial Review

The ANZCTR website will be unavailable from 1pm until 3pm (AEDT) on Wednesday the 30th of October for website maintenance. Please be sure to log out of the system in order to avoid any loss of data.

The safety and scientific validity of this study is the responsibility of the study sponsor and investigators. Listing a study does not mean it has been endorsed by the ANZCTR. Before participating in a study, talk to your health care provider and refer to this information for consumers
Trial registered on ANZCTR


Registration number
ACTRN12614001009606
Ethics application status
Approved
Date submitted
10/09/2014
Date registered
18/09/2014
Date last updated
18/09/2014
Type of registration
Retrospectively registered

Titles & IDs
Public title
A pilot study investigating the bio-availability of microRNAs from ingested fruit
Scientific title
In healthy males, does consuming fruit or other dietary sources of plant miRNAs, compared to a plant free diet, result in detectable levels of plant miRNAs in the systemic circulation.
Secondary ID [1] 285347 0
Nil
Universal Trial Number (UTN)
U1111-1154-9893
Trial acronym
Linked study record

Health condition
Health condition(s) or problem(s) studied:
This study is characterising the bio-availability of plant microRNAs from common dietary sources (fruits and grains) in normal healthy male volunteers. 293029 0
Condition category
Condition code
Diet and Nutrition 293299 293299 0 0
Other diet and nutrition disorders

Intervention/exposure
Study type
Interventional
Description of intervention(s) / exposure
This study is characterising the bio-availability of plant microRNAs following ingestion of a single 300 g serving of either kiwifruit (Actinidia chinensis var. Sungold)), apple (Malus domestica var.'Scifresh'), cooked rice (Oryza sativa L. ssp. indica) verses a control (water) in healthy male volunteers with a minimum of 7 days washout between treatment arms of the study. Small RNAs will be isolated from blood samples collected at 0, 1, 3, 6, and 24 h after the treatment meal intervention (0900 h, Day 1) and sequenced using next generation sequencing. Food diaries will be used by participants to record all food consumed 24 h prior to and during each treatment arm of the study. In addition, participants are to be maintained on a standardised plant-free diet during each treatment arm of the study starting at 1800 h on the day prior (Day -1) to the treatment meal intervention, through to the completion of the 24 h sample (0900 h, Day 2), so as to minimise contamination from other plant sources. Standardised plant-free meals will be provided to the participants for the prior evening meal (1800 h, Day -1) and the subsequent mid morning snack, lunch, afternoon snack and evening meal on the treatment day (Day 1). Participants will return the following morning (0900 h Day 2) in a fasted state to provide the final 24 h blood sample.
Intervention code [1] 290263 0
Other interventions
Comparator / control treatment
Control treatment is consuming equivalent weight (300 g) of water
Control group
Placebo

Outcomes
Primary outcome [1] 293137 0
The identification of microRNAs in plasma from specific dietary plant sources following ingestion using next generation sequencing.
Timepoint [1] 293137 0
at baseline (time 0) and at 1, 3, 6, and 24 hours after consumption of plant dietary sources of microRNA
Secondary outcome [1] 310425 0
Next Generation Sequencing (HiSeq 2000) techniques will be used to characterise the bio-availability of specific plant microRNAs. miRNAs in the treatment foods will be quantified and then compared with the levels observed in human plasma following consumption. Plasma samples will be collected at 0, 1, 3, 6 and 24 h after consumption of the treatment food and total small RNAs isolated using a Qiagen miRNeasy serum/plasma kit. Barcoded libraries of isolated small RNAs will be generated and sent to Macrogen for Next Generation Sequencing (HiSeq 2000). A range of bioinformatic tools will be used to identify miRNAs present in the samples.
Timepoint [1] 310425 0
To gain a temporal profile of the absorption of plant miRNAs, plasma samples will be collected at 0, 1, 3, 6 and 24 h after consumption of the treatment foods

Eligibility
Key inclusion criteria
Male, aged 18-55 years with a BMI between 19-29 and healthy as ascertained by self-report
Minimum age
18 Years
Maximum age
55 Years
Sex
Males
Can healthy volunteers participate?
Yes
Key exclusion criteria
Obese (BMI>29).
History of gastrointestinal disorders (Ulcerative colitis, Crohn’s Disease or Irritable Bowel Syndrome) or conditions effecting the gastrointestinal tract.
Taken antibiotics in the month prior to when the study commences.
Any medical conditions or medications.
Are on blood thinning medication (eg Aspirin).
Smoker or ex-smoker who quit within the last 6 months.
Hypersensitivities or allergies to any foods or ingredients included in the study.
Unwilling/unable to comply with study protocol.
Dislike and/or unwilling to consume items listed as study foods.
Participating in another clinical intervention trial.

Study design
Purpose of the study
Treatment
Allocation to intervention
Randomised controlled trial
Procedure for enrolling a subject and allocating the treatment (allocation concealment procedures)
Eight healthy male participants will be recruited into the study via advertisement in the local media, electronic advertisements and posters in the Auckland area. Each participant will receive all treatments in a randomized order according to a Latin square design with a minimum of 1 week washout between treatments. Allocation is not concealed
Methods used to generate the sequence in which subjects will be randomised (sequence generation)
Randomisation will be carried out using a 4 x 4 Latin Square balanced for order of presentation and carry-over effects (Williams et al., 1949, Wakeling and MacFie 1995).The design was generated in R 3.0.2 using the Williams function in the crossdes package.
Masking / blinding
Open (masking not used)
Who is / are masked / blinded?



Intervention assignment
Crossover
Other design features
The study utilises a randomised, controlled, cross-over treatment design with 4 treatment arms. The four treatment arms comprise a 300 g serving of Kiwifruit, apple, cooked rice or water for breakfast.
Phase
Not Applicable
Type of endpoint/s
Bio-availability
Statistical methods / analysis
A power analysis was performed to provide estimates of variance components using data from the published literature (Zhang et al., 2012). Based on this study, the mean and standard deviation of the total number of reads of plant miRNA per 1,000,000 mammalian reads, based on 8 samples, were 14,063 and 8,882 respectively. The 8 samples were each made up of pools of 10 individuals and it is assumed that the counts for each pool are made up as the sum of the counts for the each individual at 1/10th the volume of blood. This implies that the mean and variance of the pooled samples are an unbiased estimate of the individuals mean and variance had they been sampled at the full volume needed for individual readings, assuming that the counts are distributed in a Poisson like process.
An estimate of the within subject variation based on this total variation can be calculated using the intra-class correlation coefficient (ICC). An estimate of the ICC was not found in the literature for this type of measure, in this case it is assumed that the variation between subjects is 3 times greater than the variance within a subject i.e. ICC=0.75, and therefore the estimate of within subject standard deviation =4,041.
Based on Zhang et al., 2012, the rice fed mice had an increase from baseline to 6 hours in miRNA of approximately 40%. Assuming that the estimated mean in human is of 14,063 reads per 1,000,000 is equivalent to the 6 hours time point in mice, it would imply that the baseline mean read would be approximately 14,063/1.4=10,045. This would be an expected difference from baseline of 4,000 reads per 1,000,000.
Using the estimate of within subject standard deviation of 4,041 and a specified 4 treatment crossover design with 5 time points for each treatment, a sample of 8 subjects gives a power of 80% to detect a difference in counts of 4,000 amongst the time points using Tukey’s HSD to account for multiple testing. All power calculations were conducted using Russ Lenth’s power and sample size java applets

Recruitment
Recruitment status
Active, not recruiting
Date of first participant enrolment
Anticipated
29/08/2014
Actual
29/08/2014
Date of last participant enrolment
Anticipated
19/09/2014
Actual
8/09/2014
Date of last data collection
Anticipated
Actual
Sample size
Target
8
Accrual to date
Final
Recruitment outside Australia
Country [1] 6347 0
New Zealand
State/province [1] 6347 0
Auckland

Funding & Sponsors
Funding source category [1] 289938 0
Other
Name [1] 289938 0
The New Zealand Institute for Plant & Food Research Limited
Country [1] 289938 0
New Zealand
Primary sponsor type
Other
Name
The New Zealand Institute for Plant & Food Research Limited
Address
Plant & Food Research
120 Mt Albert Road,
Sandringham,
Auckland, 1025,
New Zealand
Country
New Zealand
Secondary sponsor category [1] 288627 0
None
Name [1] 288627 0
Address [1] 288627 0
Country [1] 288627 0

Ethics approval
Ethics application status
Approved
Ethics committee name [1] 291653 0
Northern A Health and Disability Ethics Committee
Ethics committee address [1] 291653 0
Ethics committee country [1] 291653 0
New Zealand
Date submitted for ethics approval [1] 291653 0
22/05/2014
Approval date [1] 291653 0
17/07/2014
Ethics approval number [1] 291653 0
14/NTA/72

Summary
Brief summary
Trial website
Trial related presentations / publications
Public notes

Contacts
Principal investigator
Name 51350 0
Dr John Ingram
Address 51350 0
Plant & Food Research
120 Mt Albert Road,
Sandringham,
Auckland, 1025,
New Zealand
Country 51350 0
New Zealand
Phone 51350 0
+6499257119
Fax 51350 0
Email 51350 0
[email protected]
Contact person for public queries
Name 51351 0
John Ingram
Address 51351 0
Plant & Food Research
120 Mt Albert Road,
Sandringham,
Auckland, 1025,
New Zealand
Country 51351 0
New Zealand
Phone 51351 0
+6499257119
Fax 51351 0
Email 51351 0
[email protected]
Contact person for scientific queries
Name 51352 0
John Ingram
Address 51352 0
Plant & Food Research
120 Mt Albert Road,
Sandringham,
Auckland, 1025,
New Zealand
Country 51352 0
New Zealand
Phone 51352 0
+6499257119
Fax 51352 0
Email 51352 0
[email protected]

No information has been provided regarding IPD availability


What supporting documents are/will be available?

No Supporting Document Provided



Results publications and other study-related documents

Documents added manually
No documents have been uploaded by study researchers.

Documents added automatically
No additional documents have been identified.