A Systems Biology Approach to the Interaction Between Vitamin D Supplementation and Sunlight Exposure in Brazilian Women Living in Opposite Latitudes (The D-SOL Study).
Phase: NA
Conditions: Vitamin D Deficiency
Sponsor: University of Surrey

 
Detailed Protocol D-SOL Study 
 
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 Detailed Protocol 
 
 
Protocol Title:  
A systems biology approach to the interaction between vitamin D supplementation and sunlight 
exposure in Brazilian women living in opposite latitudes (The D-SOL Study). 
 
 
Principal Investigator:  
Name: Miss Marcela Moraes Mendes (PhD researcher) 
Address: Department of Nutritional Sciences, Faculty of Health & Medical Sciences, University of 
Surrey, Guildford, Surrey GU2 7XH 
Telephone: 01483 689222 
Email: m.moraesmendes@surrey.ac.uk 
 
PhD Supervisor: 
Name: Professor Susan Lanham-New 
Address: Department of Nutritional Sciences, Faculty of Health & Medical Sciences, University of 
Surrey, Guildford, Surrey GU2 7XH 
Telephone: 01483 6896476 
Email: s.lanham-new@surrey.ac.uk 
 
Co-Supervisors: 
 
Name: Dr Kathryn Hart (Co-supervisor) 
Address: Department of Nutritional Sciences, Faculty of Health & Medical Sciences, University of 
Surrey, Guildford, Surrey GU2 7XH 
Telephone: 01483 68 6438 
Email: k.hart@surrey.ac.uk 
 
Name: Dr Laura Tripkovic (Co-supervisor) 
Address: Department of Nutritional Sciences, Faculty of Health & Medical Sciences, University of 
Surrey, Guildford, Surrey GU2 7XH 
Telephone: 01483 686429 
Email:  laura.tripkovic@surrey.ac.uk 
 
Name: Dr Patricia Borges Botelho (Collaborative supervisor) 
Address: Faculty of Nutrition, Federal University of Goiás, Goiânia, Goiás, Brazil 74605-080 
Telephone: +55 (62) 3209-6270 
Email: patriciaborges.nutri@gmail.com 
 
 
 
 
 
 
 
 
 
 
 

 
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1.0 Background & Rationale 
 
Vitamin D is derived from two sources: the skin (endogenous) and the diet (exogenous).  It is generally 
believed that the major source of vitamin D is the exposure of the skin to UVB-rays contained in sunlight. 
However, the full characterization of the contribution of diet and sunlight to optimal vitamin D status 
remains unknown. Furthermore, the response of individuals to vitamin D supplementation is known to 
be variable in all population groups since several personal and environmental characteristics may affect 
the conversion of pre-vitamin D by sunlight, such as latitude, available UVB radiation throughout the 
year, skin pigmentation, age, individual behavior, diet and genetic factors. We urgently need to define 
more closely the interaction between nutrition and genetics in response to vitamin D supplementation.  
There is a considerable lack of data in both Brazil and UK populations. (Adams and Hewison, 2010; 
(Lanham-New et al, 2012) 
 
Based on the important contribution of sunlight exposure to the production and maintenance of serum 
25OHD levels, it would be reasonable to believe that vitamin D deficiency may be a problem restricted 
to countries situated at higher latitudes. However, several studies from sunny countries have shown 
that vitamin D deficiency is a more common phenomenon, despite the abundance of sunlight in these 
locations making it a global health problem. (Lips et al, 2001; Bandeira et al, 2010). Even though Brazil 
is geographically located such that there is potentially an abundance of sunlight and therefore of vitamin 
D available throughout the year, there is still evidence of sub-optimal levels of vitamin D in the 
population. A cross-sectional study conducted in Rio de Janeiro, (latitude 22 ° S), has shown a high 
incidence of inadequate serum 25 (OH) D (68.3 %) in healthy adults (Russo et al ,2009). Much of the 
UV in sunlight is absorbed by clouds, ozone and other forms of atmospheric pollution. In higher latitudes 
as the distance from the equator increases, zenith angle is increased during winter months and 
consequently, the UV radiation reaching the earth’s surface is reduced, and therefore the cutaneous 
production of vitamin D3 is also reduced. (Lips et al 2006; Wacker and Holick, 2013). Thus, the 
cutaneous synthesis of vitamin D is expected to be greater in low- latitude regions due to greater 
exposure to UVB radiation. In the UK, there is no UV radiation of the appropriate wavelength (280mm-
310mm) from the end of October to the end of March and for the remaining months of the year, 60% of 
the effective UV radiation occurs between 11.00am and 3.00pm. Moreover, studies have shown the 
influence of skin pigmentation in reducing the UVB-mediated synthesis of vitamin D. Greater 
quantification of the effect of sunlight exposure on vitamin D status is urgently required. Therefore, we 
wish to investigate the Brazilian population, suspecting that we could be underestimating the level of 
insufficiency or deficiency of vitamin D in this population, due to the false impression that the level of 
solar radiation would suffice for people living in sunny countries. 
 
Recent studies have suggested that genetic variation could be a reasonable explanation to the 
considerable differences on vitamin D levels among population independently of latitude and sunlight 
exposure. (Valdivielso 2004) Heritability of vitamin D levels has been reported to range from 28.8% to 
68.9% in an adolescent twin study and up to 80% in a genome-wide linkage scan. (Uinterlinden 2004, 
Santos 2012). Identifying genetic variants in the form of single nucleotide polymorphisms (SNPs) has 
been one of the key methods for identifying genetic variants associated with vitamin D levels. The data 
would be helpful for identifying those who may be at risk of deficiency and targeting treatment to reduce 
the impact of vitamin D deficiency. The study will undertake a Genome-wide analysis of leukocyte gene 
expression and its correlation with differences in vitamin D status. The Vitamin D Receptor (VDR) is a 
nuclear transcription factor which controls expression of a wide range of genes, including several that 
encode cytochrome P450 enzymes and cytokines.  Human leukocytes are known to express the VDR-
encoding gene and it is therefore envisaged that differences in vitamin D responses of individuals could 
be reflected by differences in leukocyte gene expression. Therefore we propose to analyse genome-
wide transcriptomic expression within leukocytes of selected participants in order to associate specific 
signal transduction and metabolic pathways to respective vitamin D responses.   Circulating 25(OH)D 
concentrations have been shown to be strongly related to SNPs located on exons of the vitamin D 
binding protein gene: i.e. minor SNP variants of DBP-1 (rs7041,  GG®TT (17.5%) and DBP-2 (rs4588, 
CC®AA (9%), resulting in 10-13% lower circulating 25OHD levels.   
 

 
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Our results will firstly enable determination of how important (as a % contribution) diet, sunlight 
exposure and genetic factors are to vitamin D status when directly comparing, using the same 
methodology, same ethnicity populations living in a Southern Hemisphere (where there is abundant 
sunlight exposure) and in a Northern Hemisphere (where there is only sufficient sunlight exposure 
during the months April to September).  
 
The data will provide both countries with key data on whether there should be consideration of further 
revisions to dietary recommendations for vitamin D in adult populations.  This study provides good 
‘value for money’ since it draws upon previously collected data in the UK and enables a strong study 
design to be conducted in the two Countries, simultaneously.   The data will be of great interest to the 
field of Nutritional Sciences by the provision and formulation of data, within a short period of time, 
examining the interaction of diet and the environment on vitamin D status and its functional 
consequences. The ‘systems level’ approach will enable us to identify differences in gene expression 
and whether this explains why some individuals are ‘good’ responders or ‘poor’ responders to vitamin 
D supplementation (Wang et al, 2010).  This has never been done before in the field, by examining two 
population groups living in different countries but following identical study designs. Also, to our 
knowledge, this is the first randomized control trial to analyse the effect of vitamin D supplementation 
in Brazilian women, either living in the UK or in Brazil.  
 
 
2.0 Objectives & Design 
 
 
2.1 Primary Objectives: To examine the response (in vitamin D status) to vitamin D 
supplementation and sunlight exposure, and its influencing factors within and between 
Brazilian female adults living in the UK and Brazilian female adults living in Brazil. 
 
2.2 Secondary Objectives 
  
(i) Analyze the difference regarding time and intensity of sun exposure between Brazilian 
women living in Brazil and Brazilian women living in the UK. 
(ii) Determine the prevalence of inadequate dietary Vitamin D intake in these women. 
(iii) Determine the prevalence of insufficient/deficient levels of vitamin D in these women. 
(iv) To elucidate the association between Vitamin D status and markers of calcium of calcium 
metabolism. 
(v) Examine the influence of latitude on vitamin D optimal levels. 
(vi) Analyze the influence of skin pigmentation on vitamin D optimal levels. 
(vii) Analyze if response to Vitamin D supplementation is dependent on initial vitamin D levels. 
(viii) 
Investigate the genetic and enzymatic mechanisms underlying the adolescents’ 
response to vitamin D supplementation via genotyping for polymorphisms related to vitamin 
D metabolism and comparing the high and low responders to supplementation (Genetic 
Sub-Study). 
 
 
2.3 Hypothesis 
 
The study hypothesis that, during winter, vitamin D supplementation is required to obtain Vitamin D 
optimal levels in Brazilian women residing in the UK as well as Brazilian women residing in Brazil and 
that this response is dependent on the initial levels, being also influenced by sunlight exposure, skin 
pigmentation and polymorphisms of the vitamin D receptor gene. 
 
H1: Dietary intakes of vitamin D are too low and sunlight exposure is insufficient for optimal vitamin D 
status in Brazilians living in the UK and Brazilians living in Brazil. 
 
H2: The response of subjects to vitamin D supplementation is dependent on baseline 25OHD levels.  
 

 
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H3: Response to supplementation is influenced by sun light exposure. 
   
H4: Skin pigmentation is an influential factor on vitamin D optimal levels. 
 
H5: There will be strong genetic influences on 25OHD changes in response to vitamin D 
supplementation; these will be similar in both groups.  
 
 
3.0 Experimental Design 
 
Two controlled, randomized, double-blind clinical trials will be developed and undertaken (one in Brazil 
and the other in the UK) with an intervention period of 12 weeks. 
 
A questionnaire to screen for the relevant inclusion and exclusion criteria (see section A) will be 
administered in order to select 80 Brazilian female subjects, aged 20 to 59 years, in each of the two 
countries. The women selected will be randomly divided into two groups: Placebo Group and 
Supplemented Group, the latter will receive 600UI UI of vitamin D.  
The first clinical trial will run in the UK from October 2016 to March 2017 (autumn-winter) and then the 
second will run in Brazil from April 2017 to September 2017 (autumn-winter). After the analysis of the 
effect of vitamin D supplementation compared to placebo, the results obtained in the supplemented 
group will also be analysed according to the genotypes for SNPs in the VDR gene. 
 
We have chosen 12 weeks (3 months) as the supplementation period as we consider this to be ample 
time for 25OHD levels to rise (half-life of 25OHD is 3 weeks) and this has been demonstrated to be an 
effective timeframe for supplementation of vitamin D in previous studies.   
 
We have chosen 600IU to enable the study to be relevant to the new IOM US recommendations for 
vitamin D, which are currently the reference document used in Brazil, even though the Brazilian 
nutritional table remains with the daily intake recommendation of 200 IU, which was the IOM 
recommendation in 1997.  Compliance will be checked by compliance interviews during the trial and by 
empty packaging count.   
 
We will genotype vitamin D related genes in DNA isolated from the study blood samples, DNA will be 
extracted and vitamin D polymorphisms will be determined by our genetic labs. These will include known 
candidate variants (VDR, vitamin D binding protein, CYP2R1, CYP27A1, CYP27B1, and CYP24A1), as 
well as variants identified by the ongoing genome-wide association analyses on 25OHD.  We will 
construct a genetic risk score based on these polymorphisms. The Genome-wide expression profiling 
of RNA from leukocytes will focus on 48 participants (each at two sampling time points) that represent 
equally both groups, encompassing the best and worst supplementation-responders in each group. We 
will identify the complete spectrum of genes that are, by statistical criteria, either significantly up-or 
down-regulated (i.e. differentially expressed) between groups. The major comparison will be between 
the ‘good’ and ‘poor’ responders in each group; gene expression prior to supplementation will be 
compared to   after supplementation for each subject analysed.  
 
 
3.1 Sample Size and randomisation 
 
A total of 80 subjects (at 90% power) are required for recruitment into the 600 IU vitamin D group vs 
placebo group for the RCT to be conducted in the UK and a total of 80 subjects (at 90% power) are 
required for recruitment into the 600 IU vitamin D group vs. placebo group for the RCT to be conducted 
in Brazil.  This will enable us to detect a 0.6 SD size effect at 90% power in serum 25OHD levels 
between placebo and 600 IU in UK-dwelling Brazilian populations and Brazilians living in Brazil.  These 
study numbers of 40 subjects for the vitamin D group and n 40 for the placebo group includes a 20% 
drop-out rate factored in.  
 

 
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3.2 Recruitment 
 
Participants in the UK will be recruited through advertisement among local Brazilian societies/groups 
within Surrey and London. Informative posters will be positioned around the University of Surrey and 
commercial centres (with a focus on Brazilian themed places), with permission of the owner.  
 
As recruitment method, we will ask, after clearly explaining our intentions of divulgating the study, 
Brazilian institutions in the UK, such as the Brazilian Consulate and the Brazilian Researchers 
Association (ABEP-UK) to circulate a recruitment letter to their contact list of Brazilians living in the UK.  
If we succeed in this arrangement, this letter will be send directly to those in the lists by the institution, 
therefore we will not need to have access to people’s contact details. 
 
Social media sites such as Facebook will be used to publicise the study: the poster or flyer will be 
inserted as a photo with permission of the administrator of the Facebook pages.  
Facebook pages we intend to target :  
Science Without Borders UK students: https://www.facebook.com/groups/489494047812445/ and 
https://www.facebook.com/groups/csf2014/ 
Brazilians in London: https://www.facebook.com/groups/301278086667948/  
Brazilian Students in the UK Association (ABEP): https://www.facebook.com/ABEPUK/?fref=ts  
General Consulate of Brazil in London: https://www.facebook.com/cglondres/info/ 
 
Participants in Brazil will be recruited from the general public within the city of Goiânia through posters 
to be positioned around the Federal University of Goiás and commercial centres, with permission of the 
owner.  
Social media sites such as Facebook will be used to publicise the study: the poster or flyer will be 
inserted as a photo with permission of the administrator of the Facebook pages.  
Facebook pages we intend to target:  
Federal University of Goias: https://www.facebook.com/universidadefederaldegoias/?fref=ts 
Faculty of Nutrition, Federal Uni of Goias: https://www.facebook.com/fanut.ufg.5?fref=ts 
Faculty of Nutrition Postgrads, Federal Uni of Goias: 
https://www.facebook.com/mestrado.fanut.7?fref=ts 
 
The recruitment end date for the UK subjects will be 14th December 2016. Recruitment end date for the 
Brazilian subjects will be the 13th June 2017. 
 
Subjects will be reimbursed for their travel expenses. 
 
 
3.3 Selection & Withdrawal of Participants  
 
3.3.1 Inclusion Criteria 
 
 
Brazilian nationality  
 
Female 
 
Aged 20-59 years 
 
 
3.3.2 Exclusion Criteria 
 
 
Currently receiving treatment for medical conditions that are likely to affect vitamin D 
metabolism ( osteoporosis therapy, anti-estrogens treatment, antiepileptic drugs, breast-cancer 
treatment) 
 
Hypercalcaemia (>2.5mmol/L) – assessed and excluded at baseline 

 
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 
Regular use of sun-beds 
 
Having a holiday trip one month prior to commencing the study or plans for a holiday trip for 
more than 4 weeks out of the country of residence within the study period.  
 
Use of vitamin supplements containing vitamin D (if the prospective participants agrees to stop 
Vitamin D supplementation to join the study, a wash-out period of 8 weeks prior to commencing 
the trial would be acceptable).  
 
Pregnant or planning a pregnancy during the study period.  
 
If a participant is subsequently found to be ineligible for the study their screening questionnaire will be 
destroyed due to the questionnaire containing sensitive information. 
 
 
3.3.3 Withdrawal 
 
All participants will be notified during the consenting process that they are free to withdraw from the trial 
at any time, without giving a reason. 
 
Participants will be withdrawn from the trial by the Principal Investigator if: 
1. The participant develops a medical condition or becomes pregnant either prior to entering 
the study or during, which may adversely affect the outcome of the study. 
2. It is clearly demonstrated that the participant is non-compliant completing study activities 
and the control procedures requested of them. 
3. A participant suffers an adverse event, which will be reviewed and recorded at each visit 
and at each phone call, following the immediate discontinuation of the participant during 
the visit, if necessary. 
 
All data prior to subject withdrawal will be used in analysis; unless the participant specifically requests 
that their data is not to be used. Withdrawn participants will not be replaced as an anticipated drop-out 
rate of 20% has been accounted for in the recruitment targets. 
 
 
4.0 Trial Procedures 
 
4.1 Trial visit activities 
 
During this study the subjects will be asked to visit the Clinical Investigation Unit, FHMS, University of 
Surrey in the UK or the Research Clinic, Faculty of Nutrition, Federal University of Goias in Brazil, on 
two occasions, at the beginning of the study for baseline measurements and at the conclusion of the 
study. 
We will examine four to eight fasted subjects per study morning.   
Trial visits will last approximately 45-60 minutes each and take place in the morning (7am-11am). 
Participants will be offered refreshments at the end of their appointment. 
 
 
Consenting and Screening 
 
If participants wish to be screened for participation in the study, they will receive the Participant 
Information Sheet and then be checked against the study inclusion and exclusion criteria using the 
‘Screening Questionnaire’ (Section A), to be administered by a member of the D-SOL Research Team 
by phone or self-reported by email. 
 
Baseline visit  
 
If eligible, participants will be invited for the baseline visit. At this visit, they will first be given time to 
discuss the Participant Information Sheet and any questions they may have regarding the study. 

 
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Informed consent will be discussed and participants will be asked to sign the consent form, and will be 
offered a copy to keep for themselves. 
 
 
Health and Lifestyle questionnaire to be administered by a member of the D-SOL Research 
Team. 
 
Anthropometrics and blood pressure measured, and fasted blood sample taken (serum 25OHD 
levels, 1,25-dihydroxy vitamin D, serum calcium, albumin, parathyroid hormone, C-terminal 
telopeptide (CTX) ≈25ml) with an additional ≈10 ml for genetic profiling and ≈15 for storage for 
future measurements of nutritional markers. 
 
pQCT scan of the non-dominant forearm for UK trial or DEXA scan for Brazil trial. 
 
Bioelectrical impedance analysis (BIA) for body composition. 
 
Provision of randomly assigned daily supplement (30 days’ supply), food diaries and dosimeter 
(to be returned via SAE provided) and sunlight exposure diary to be returned at 12 week visit. 
Follow-up appointment details arranged (including interim visit telephone appointments and for 
re-supply of supplement). 
 
 
Final visit 
 
 
Final adverse event/compliance interview completed with investigator. 
 
Daily outdoor exposure diary received from participant and check for consistency at visit. 
 
Anthropometrics and blood pressure measured, and blood sample taken (serum 25OHD levels, 
1,25-dihydroxy vitamin D, serum calcium, albumin, parathyroid hormone, C-terminal 
telopeptide (CTX) ≈25ml) with an additional ≈10 ml for genetic profiling and ≈15 for storage for 
future measurements of nutritional markers. 
 
Bioelectrical impedance analysis (BIA) for body composition. 
 
Dosimeter and 4-day food diary received from participant (sent to participant prior to 
appointment). 
 
 
DNA profiling procedure - After 12 weeks RCT 
 
 
Selection 
of 
48 
participant 
samples 
encompassing 
the 
best 
and 
worst 
supplementation-responders in each group, subject to previous consent form 
participant. Vitamin D related genes will be genotyped in DNA isolated from the study 
blood samples, DNA will be extracted and vitamin D polymorphisms will be determined 
by University of Surrey’s genetic labs. 
 
A trained phlebotomist will take the blood samples required as part of the trial protocol. Medical cover 
will be available at all times.  
 
Throughout the duration of the trial, the participants will be contacted via telephone on a fortnightly basis 
to discuss any issues with any adverse event and compliance and to maintain good communication 
with the participants. (Please see AE reporting form). In the case of a serious adverse event (SAE) this 
will be recorded (please see SAE reporting form) and we will report it to both the Sponsor (University 
of Surrey) and the Surrey Ethics Committee. 
The final interview will be completed at the final study visit. Participants have also be asked to return 
any supplements that were missed to confirm compliance. 
 
For University of Surrey participants: A peripheral quantitative computed tomography (pQCT) scan was 
performed on the participant's non-dominant forearm at the baseline visit, to measure volumetric bone 
mineral density at the 4% and 66% radial site. This will allow for separate measurements of trabecular 
vBMD and trabecular area (4% site) and cortical vBMD and cortical area (66% site), as well as strength 
strain index, a measure of bone strength. pQCT also measures bone geometry alongside bone density. 
Therefore the muscle cross sectional area can be determined, which is a measure of muscle force, to 
which bone strength is adapted to. One scan was performed at baseline only and effective exposure 
doses were between ~1.5-1.8uSv. 
 

 
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For Federal University of Goiás participants: Body composition (absolute and relative amount of lean 
and fat mass), whole body mineral density and lower spine and femur bone mineral density was 
measured with the use of DEXA (located at the Nutrition Clinic based at the Federal University of Goias), 
at baseline only. Two scans were performed at baseline for each participant: one to assess the whole 
body mineral density and body composition, and the other to specifically assess fracture risk by 
scanning the spine and femoral head. Effective exposure doses for theses scans are ~8uSv and ~4uSv 
respectively. 
 
Results of the body composition, vitamin D status and dietary intake from the self-reported food diaries 
will be made available to the subjects upon request. The results from the blood analysis will be reported 
to the subject if there are any health concerns raised. If the results are within healthy ranges the 
participants will not be contacted unless they specifically request for this information.  The investigators 
will not be contacting their GPs if there are any concerns raised in the study however we will stress that 
they should contact their GP themselves to discuss the results we found.  
 
 
5.0 Benefits and risks of participants in the D-SOL Study 
 
Results of the body composition, vitamin D status and dietary intake from the self-reported food diaries 
will be made available to the subjects upon request.  
 
We do not anticipate identifying anything of concern via either the blood tests or bone scan, as we’re 
not measuring any markers that would actually indicate any serious pathology is occurring. A letter 
informing that the participant is on the trial will be sent the participant’s GP. A response from the GP 
will not be required unless the GP had concerns and no results will be sent to them afterwards. If the 
results are within healthy ranges the participants will not be contacted unless they specifically request 
for this information. If abnormal results came back for a participant, they will be offered support in 
approaching the GP, all the information regarding their test results, details on the intervention, etc, will 
be made available.  
 
Due to the trial being food based, the risk of side-effects is minimal. However, gastrointestinal discomfort 
may occur following supplement consumption. 
 
A blood sample must be taken at each trail visit, including screening, and due to the nature of the 
procedure, some light bruising may occur. Occasionally, fainting in some individuals can occur relating 
to venepuncture. TO help reduce the risk of this, participants will have their blood sample taken either 
whilst they are supine on a bed or reclined on a chair that has the capacity to be adapted quickly to 
allow the participant to lie supine safely if they do become unwell. 
 
pQCT and DXA scans use a low level of radiation (much lower than standard X-ray examination) to 
which participants will be exposed. The amount of radiation absorbed from the scan is very small and 
similar to the radiation we receive from the environment. 
 
Although health problems linked to vitamin D are very rare, there is some evidence that vitamin D 
supplementation can cause blood calcium levels to go higher than normal in some people who already 
have high blood calcium levels. Upon commencing the study, all participants will have their blood 
calcium levels checked to ensure that the vitamin D would be unlikely to cause any problems. 
 
6. Ethics and regulatory approval 
 
The trial will be conducted in compliance with the principles of the Declaration of Helsinki (2008), the 
principles of GCP and in accordance with The Medicines for Human Use (Clinical Trials) Regulations 
2004 and Amended Regulations 2006. 
 

 
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This protocol and supporting documents will be submitted for review by the University of Surrey Ethics 
Committee and the Federal University of Goias, Brazil. Annual progress reports and a final report will 
be submitted to the ethics committees as defined in their respective regulations. 
 
 
 
7.0 Study Evaluation and Statistical Analysis 
 
Statistical analysis will be undertaken with support from the University of Surrey statistical department. 
Data will be checked for normality using appropriate testing. Appropriate parametric/non-parametric 
analysis will be applied.   
The database will be stored and analysed using SPSS ® version 13.0. 
The descriptive analysis, including mean ± standard deviation, median and lower limit and higher will 
be held for all quantitative variables Data will be checked for normality using appropriate testing. 
Appropriate parametric/non-parametric analysis will be applied. Student t test or Mann-Whitney test, 
depending on the distribution of data, will be applied to evaluate differences between supplemented 
and placebo. For analysis between Brazil and UK groups an ANOVA test will be undertaken. Linear 
correlation, Pearson or Spearman, will be calculated in accordance with the presence / absence of 
normal distribution, respectively, between vitamin D intake, sun exposure, skin pigmentation and 
vitamin D serum concentration. Multiple linear regression analysis will be performed to determine the 
variants that influence blood concentrations of vitamin D. In order to determine whether supplemental 
response differs between genotypes, these will be separated into wild homozygotes, heterozygotes and 
homozygotes for the variant. In this approach, the results will be evaluated with the test ANOVA or 
Kruskal-Wallis, in cases of variables with normal distribution or not, respectively. To describe the 
relationship between food intake aspects and features biochemical independent of energy intake, intake 
values of nutrients of interest will be adjusted to energy in accordance with the method.  
Significance considered as standard will be 5%. 
 
 
8.0 Data Handling 
 
The Principal Investigator will act as custodian for the trial data. The following guidelines will be strictly 
adhered to: 
 
 
Participants data will be completely anonymised 
 
All anonymised data will be stored in a secure location on the University’s servers and on a 
password-protected computer these will be in line with best practice as recommended in the 
University of Surrey Research and Information Governance policies.  
 
All trial data will be stored and archived as indicated by The Medicines for Human Use (Clinical 
Trials) Amended Regulations 2006. 
 
When a subject does not meet the inclusion criteria, all questionnaires and collected data will 
be destroyed. 
 
As part of the collaboration agreement with the Federal University of Goias, Brazil, all data 
collected in both countries will be shared between the two institutions. 
 
 
9.0 Publication policy 
 
The results of the study will be reported and disseminated to the scientific community via peer-reviewed 
journals and international conferences. The general public will be engaged via the release of results to 
the local and national media, relevant charities and community networks and an invited talk at the 
University.  
 
10.0 Finance 
 

 
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Full funding to conduct the D-SOL Study is provided by the Science Without Borders Program through 
the National Counsel of Technological and Scientific Development of Brazil (CNPq).  
 
11.0 Signatures 
 
Principal Investigator:  
 
 
 
 
Date: 
Miss Marcela Moraes Mendes   
 
 
 
17 January 2016 
 
REFERENCES 
 
 
WANG TJ, et al. Common genetic determinants of vitamin D insufficiency: a genome-wide association 
study. Lancet. v. 376 n.9736, p. 180–8, 2010. 
 
ADAMS, JS AND HEWISON, M. Update in Vitamin D. J Clin Endocrinol Metab. v.95, n. 2, p. 471–478, 
2010. 
 
BANDEIRA, F et al. Vitamin D deficiency and its relationship with bone mineral density among 
postmenopausal women living in the tropics. Arq Bras Endocrinol Metab, 
São Paulo, v. 54, n. 2, 2010. 
 
LIPS, P. Vitamin D deficiency and secondary hyperparathyroidism in the elderly: consequences for 
bone loss and fractures and therapeutic implications. Endocr Rev. , v. 22, n. 4, p. 477-501, 2001. 
 
LANHAM-NEW S et al. Comparison of vitamin D2 and vitamin D3 supplementation in raising serum 25-
hydroxyvitamin D status: a systematic review and meta-analysis. Am J Clin Nutr, v. 95, n. 6, p. 1357-
1364, Jun 2012. 
 
SHEA, MK et al. Genetic and non-genetic correlates of vitamin K and D. European journal of Clinical 
Metab. 63(4), 2009. 
 
SANTOS, BR et al. Vitamin D deficiency in girls from South Brazil: a cross-sectional study on prevalence 
and association with vitamin D receptor gene variants. BMC Pediatrics. v. 12, n. 62, 2012 
 
UITTERLINDEN AG, et al. Genetics and biology of vitamin D receptor polymorphisms. Gene. v. 338, 
n. 2, p. 142-56, 2004. 
 
 VALDIVIELSO JM, FERNANDEZ E. Vitamin D receptor polymorphisms and diseases. Clin Chim Acta. 
v. 371, n.1, 2006. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

 
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SECTION A 
 
THE D-SOL STUDY 
SCREENING QUESTIONNAIRE 
 
The purpose of this questionnaire is to assess whether you are suitable to take part in the study and 
that it is safe for you to do so. We would be grateful if you would answer the following questions, even 
if you are still not sure if you wish to take part in our study. Please answer the questions as honestly 
and accurately as you can and remember there are no right or wrong answers to the questions. Your 
answers to the questions on this questionnaire will also be kept completely confidential.  
If you feel uncomfortable answering any of the questions on this questionnaire you do not have to 
answer the question. Please ask one of the D-SOL Research Team if you would like help answering 
any of the questions. 
 
 
 
Name:                                                   
 
DOB:        
 
 
    Age:  
  Gender:    
 
Address: 
 
 
 
Contact telephone number: 
Place of birth:            Nationality:  
Mother’s nationality:                                             Father’s nationality: 
Email address: 
Preferred method of contact:       
Phone                   Email                    Post 
 
 
 
 
 
 
 
Height (cm): ………………………. 
Weight (kg): ………………… 
 
 
Are you currently receiving treatment for any medical conditions?  
 Medical condition: …………………………… 
Treatment: ………………………. 
/
/
CONTACT DETAILS 
HEALTH AND LIFESTYLE  
Yes 
No 

 
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Are you on any medication prescribed by your GP or any other health care provider? 
 
 
     If yes, please specify: ………………………………………………………. 
 
1. Do you live in the UK?  
If yes, please state the date you arrived: 
 
2. Please tick all medical conditions that apply: 
 
Prior/ present history of coronary heart disease, angina, heart attack or stroke 
 
Prior/present history of Type 1 and Type 2 Diabetes. 
 
Prior/present history of Thyroid disease 
 
Prior/present history of osteoporosis, osteopenia or other musculoskeletal disease 
 
Prior/present history of haematological disease (except mild anaemia) 
 
Prior/present history of malignancy 
 
Prior/present history of a gastrointestinal disorder, such as Crohns Disease, Coeliac 
Disease or Irritable Bowel Syndrome. 
 
Prior/present history of liver or kidney disease. 
 
 
3. Do you have any allergies or food intolerances? 
If yes, please specify: ……….………………………………………………………… 
………………………………………………………………………………………… 
 
4. Do you regularly take vitamin supplements containing vitamin D? 
 
If yes please specify:  
How many months you have been taking the supplements?................................ 
The dose of the supplements (if known): ……………………………………… 
 
 
The brand of the supplements (if known):  ……………………………………. 
 
5. Are you currently on a weight-reducing diet or other dietary restrictions (except 
vegetarianism)? 
If yes, please provide details: ………………………………………………………….. 
6. Have you been abroad on holiday during the past 6 months? 
If yes, please specify where this was and the month this holiday was taken: 
Country of visit…………………………………………………….. 
Month/year…………………………………………………….. 
 
 
Country of visit…………………………………………………….. 
Month/year…………………………………………………….. 
 
 
 
Yes 
No 
Yes 
No 
Yes 
No 
Yes 
No 
Don’t know 
Yes 
No 
Yes 
No 
/
/

 
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6. Are you planning any holidays abroad during the next 12 months? 
If yes, please specify where this visit is planned to be and when this holiday will be 
taken: 
Country of visit…………………………………………………….. 
Month/year 
……………………………………………………..  
Country of visit…………………………………………………….. 
Month/year 
……………………………………………………..  
7. Do you use sunbeds? 
If yes, please state how often you use them: 
 
 
 
Once a week 
Once a month 
 
 
More than 6 times per year 
 
 
Less than 6 times per year 
 
 
Occasionally  
8. Are you currently pregnant or planning a pregnancy during the next 12 months? 
 
9. Are you currently breastfeeding? 
 
 
 
 
10. Are you in menopause? 
 
 
 
 
 
 
We are planning to see participants in the morning between 8.00am and 12.00 noon. 
Are there any particular day or days which would be best for you? 
 
 
 
 
 
 
 
 
 
 
 
 
 
Yes 
No 
Yes 
No 
Yes 
No 
Yes 
No 
 

 
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Thank you for taking the time to complete this questionnaire. 
 
Participant signature: __________________________________________________ Quest. No:  
Date: ___/___/_____