Bioavailability of Proteins From Plant Based Diets
Schlüsselwörter
Abstrakt
Beschreibung
This study is divided in 4 phase. First Phase: Agriculture (Year 1) Site description: The experiment will carried out at the Experimental Farm of São Paulo's Agency for Agribusiness Technology (APTA) in Ribeirao Preto, Brazil, at the geographic coordinates of 21°12'26" S e 47°51'48" N and average altitude of 646 m. The annual rainfall is about 1427 mm, with average maximum and minimum temperature of 25°C and 19.3°C, respectively. The soil is classified as Oxitol with glay texture. The bean crop (Phaseolus vulgaris), will be grown at the 0,50m between rows with 12 seed per linear meters and at the 2 weeks after anthesis (flowering) the water supply will be amended with deuterium oxide to experimental development. The grain produced will be harvested to experimental analysis and consumption. The experimental area will have 5.000 m2 with common bean crop to produce enough quantity of grains to the experiment running. Seeds will be dried in a scale of 1 to 10 kg of dry and selected seeds.
Nutrients Composition Analysis (Year 1) Six samples of 100g each will be separated for nutrients composition analyses, that will be performed at the Animal Nutrition Laboratory of the Animal Science Institute of the University of São Paulo, Brazil. Right after harvesting, the food samples will be weighted and dried in a stove with forced air circulation at 65 degrees Celsius, during 72 hours. After this, samples will be ground in a mill with 2 mm hole sieve, and a subsample will be taken for the analysis of dry mass, protein, ether extract, mineral mass, energy, calcium and phosphorus, according to Official Methods of Analysis (AOAC) (1996). Fibre will be analysed in neutral and acid detergent according to reference Silva and Queiroz, 2005, and selenium will be analysed according to Olson et al., 1975. Energy analysis will be performed by a calorimetric bomb and fat by the extraction according to reference Bligh and Dyer, 1959, at the Nutrients Composition Laboratory of the Ribeirão Preto Medical School.
Second phase (pilot study), Nutrition Institute (Year 2) Subjects Fifteen adult volunteers will be recruited for method validation. Inclusion criteria
- Healthy, non-pregnant adults 18-65 years of age
- Not taking any medication, including NSAIDs
- Not taking antibiotics within 4 weeks of study
- BMI within 18-35 range
- Stable weight Exclusion criteria
- Children and elderly (over 65)
- Pregnancy
- Diagnosed with non-communicable or communicable disease
- Being under restrictive diet
- Antibiotics within 4 weeks of study
- Medications within 4 weeks of study Study design After approval of project by local ethics committee and obtaining signed informed consent, volunteers will be offered a test meal with milk protein. The test protein in the first place will be whole milk protein, produced with a low level deuterium enrichment within the study. This will permit the Carbon-13 single cell protein to be characterised against a well-studied animal protein known to possess high digestibility. Tracer Amino Acid (AA) appearance in the circulation (blood; saliva, urine and breath will also be sampled) and the relative appearance of test and reference protein will be taken as a measure of relative bioavailability which can be converted into an absolute measure once the reference protein has been characterised. The basic measurement will be of stable isotope enrichment in individual amino acids.
Phase 1 pilot study, Nutrition Institute. Fifteen adult volunteers will be recruited for method validation.
Third Phase is in 15 children (1-3 years, with different nutritional status) and Fourth phase is in 15 elderly (more than 65 years, with sarcopenia or not sarcopenia).
Test meal 1: Beans soaked for 3 h, water discarded and beans cooked for 20 min in pressure cook.
The test meal will be separated by the use for two weeks of unlabelled beans. The same approach as used in the pilot study of milk protein will be adopted to study the bioavailability of grain legumes through simultaneous comparison with a modest quantity carbon-13 enriched single cell protein combined in the same test meal.
All groups will be analysed food intake assessment, total urinary nitrogen and enrichment of deuterium in protein. Four 24h Recall Questionnaires will be applied to each volunteer during the periods of intake of the test meal, in order to verify the protein and calories consumption. Volunteers will be asked to keep their usual intake, to avoid changes in food consumption. Total urinary nitrogen of the samples will be analysed by the Kjeldahl method with ammonium micro diffusion. Compound specific isotope enrichment in amino acids will be analysed in saliva, urine, breath and blood samples. Carbon-13 in breath carbon dioxide (CO2) and Deuterium in urine enrichment will be analysed in Mass Spectrometry of Medicine School of Ribeirão Preto (FMRP-USP), Brazil, SP.
Termine
Zuletzt überprüft: | 04/30/2018 |
Zuerst eingereicht: | 02/22/2018 |
Geschätzte Einschreibung eingereicht: | 04/03/2018 |
Zuerst veröffentlicht: | 04/09/2018 |
Letztes eingereichtes Update: | 05/02/2018 |
Letztes Update veröffentlicht: | 05/03/2018 |
Tatsächliches Startdatum der Studie: | 06/30/2018 |
Geschätztes primäres Abschlussdatum: | 08/22/2018 |
Voraussichtliches Abschlussdatum der Studie: | 12/30/2020 |
Zustand oder Krankheit
Phase
Armgruppen
Arm | Intervention / Behandlung |
---|---|
adults Select the individual:
Healthy, non-pregnant adults 18-65 years of age, not taking any medication, including NSAIDs, not taking antibiotics within 4 weeks of study, BMI within 18-35 range and stable weight.
Exclusion criteria: children and elderly (over 65), pregnancy, diagnosed with non-communicable or communicable disease, being under restrictive diet, antibiotics within 4 weeks of study, medications within 4 weeks of study. They will eat beans for 5 days. The beans will be soaked for 3 h, water discarded and beans cooked for 20 min in pressure cook. And in the end for the week, adults will be eat the bean enrichment of deuterium for availability protein, collected urine, saliva and blood. | |
Children Fifty children will be recruited from ages varying from 1 to 3 years and of both sexes. Children who use medications, who do not habitually consume beans, will be excluded and when there is no explicit written authorization from the parents or guardians.
All the children with different status of nutrition will be eaten the beans. They will eat beans for 5 days. The beans will be soaked for 3 h, water discarded and beans cooked for 20 min in pressure cook. And in the end for the week, children will be eat the bean enrichment of deuterium for availability protein, collected urine or saliva depend on the best biological samples of pilot study (adult). | |
Elderly Fifty individuals of both sexes will be recruited, previously evaluated by complete clinical / laboratory examination and medical history. The elderly will be excluded from antiinflammatory, chemotherapeutic, corticoid, allergy and bean aversion or antibiotic therapy. These drugs could interfere with protein bioavailability. They will eat beans for 5 days. The beans will be soaked for 3 h, water discarded and beans cooked for 20 min in pressure cook. And in the end for the week, elderly will be eat the bean enrichment of deuterium for availability protein, collected urine or saliva depend on the best biological samples of pilot study (adult). |
Zulassungskriterien
Altersberechtigt für das Studium | 18 Years Zu 18 Years |
Studienberechtigte Geschlechter | All |
Probenahmeverfahren | Non-Probability Sample |
Akzeptiert gesunde Freiwillige | Ja |
Kriterien | Inclusion Criteria: - Healthy, non-pregnant adults 18-65 years of age - Not taking any medication, including NSAIDs - Not taking antibiotics within 4 weeks of study - BMI within 18-35 range - Stable weight Exclusion Criteria: - Children and elderly (over 65) - Pregnancy - Diagnosed with non-communicable or communicable disease - Being under restrictive diet - Antibiotics within 4 weeks of study - Medications within 4 weeks of study |
Ergebnis
Primäre Ergebnismaße
1. Bioavailability protein using deuterium [It will be collected for 1 day]
Sekundäre Ergebnismaße
1. Bioavailability protein using carbon 13 [It will be collected for 1 day]
2. Nutritional composition [through study completion in 1 week]
Sonstige Ergebnismaßnahmen
1. Total urinary nitrogen [It will be collected for 1 day]