Health Impacts of Sustainable Ingredient Selection in the Food and Drink Industry - ALTERNATIVE PROTEIN STUDY
Health Impacts of Sustainable Ingredient Selection in the Food and Drink Industry - ALTERNATIVE PROTEIN STUDY
Summary

 This research will examine the nutritional effects of supplementing diets with alternative
 plant sources of protein with potential to be grown in Scotland. In particular, we will
 assess the potential of these plant protein sources to complement diets in which the
 predominant source of protein is meat. These protein sources will include plants from the
 Fabaceae, Cannabaceae and Polygonaceae families, which could be used as the basis for the
 development of new foods. The research will assess if these plant proteins can deliver a
 comparable alternative to meat based diets.

 It is anticipated that the results could encourage increased consumption of plant products
 which would be favorable for consumers shifting away from animal-derived proteins for health
 and/or environmental reasons.

 Hypothesis: Consumption of protein rich plants could be a sustainable and healthy choice for
 partial replacement in predominantly meat based diets.

 Objective: The objective of this acute study is to assess satiety, postprandial effects,
 metabolite bioavailability and metabolism of single alternative proteins from a shortlist
 including buckwheat, fava beans, lupin, pea, and hemp in comparison with red meat.

 Study protocol

 Aims: To assess the impact of a pea, fava bean, lupin, hemp, buckwheat in comparison with
 meat in healthy people on:

 -  Biomarkers of satiety as measured by gut-related hormones and subjective appetite using
 visual analogue scales, specifically to be collected during the meal over a three hour
 period, every 30 minutes. The energy intake measured for each volunteer after an ad
 libitum lunch (five hours after test meal).

 -  Biomarkers of CVD risk including total cholesterol, low density lipoproteins (LDL), high
 density lipoproteins (HDL), triglycerides and non-esterified fatty acids (NEFA).

 -  Assessment of peripheral glycaemic control, fasting glucose, area under the curve
 combined with insulin data.

 -  Plasma and urine markers of important phytochemical and protein metabolites will be
 quantitatively analysed to determine the systemic availability, in vivo concentrations
 and excretion times.

 -  Volunteer views towards diets rich in plant proteins will be assessed.
Background

 Recent scientific evidence suggests that diets largely based on plant foods, such as
 well-balanced vegetarian diets, have potential to improve health, in contrast with
 predominantly meat-based diets. However, many consumers may not wish to exclude meat from the
 diet, despite current advice to reduce, in particular, processed meat. One way to achieve
 this is to increase consumption of plant-based protein components in the diet.

 Making a comparison of the nutritional content of high protein plants (i.e. those with > 20 g
 protein/100 g dry weight), several candidate sources were identified. In particular, these
 belong to the Fabaceae family and include pulses (green peas, fava beans, lupin seeds) and
 certain common bean species. Species from plant families other than the Fabaceae generally
 have less protein. An exception is hemp seed (Cannabaceae family), which can have a protein
 content around 40g protein /100g dry weight, is a source of fibre, and has favourable fatty
 acid and amino acid profiles. Other families of interest include the Amaranthaceae species
 (Quinoa), Polygonaceae (Buckwheat) and the Poaceae (Wheat, Barley and Oats). Protein content
 of these pseudocereal and cereal crops ranges from 12-20 g protein /100 g dry weight.
 Although lower in protein content compared with the Fabaceae species, they are generally rich
 in fibre and consequently will impart additional benefits to a high-protein plant based diet.

 According to the Scottish Health Survey (2009), nearly two-thirds of men (66.3%) and
 approximately half of women (58.4%) were overweight. Obesity is a major risk factor for
 metabolic syndrome, which is linked to conditions such as type II diabetes mellitus (T2DM).
 Having impaired glucose tolerance doubles the chance of morbidity and mortality from
 cardiovascular disease (CVD). CVD is the leading cause of death and accounts for nearly 30%
 of deaths worldwide each year. Clinical studies have shown that regular consumption of pulses
 can reduce blood levels of total cholesterol and low density lipoprotein (LDL).

 The recent meta-analysis of randomized controlled trials provides the strongest evidence to
 date that non-soya legume consumption lowers serum total and LDL cholesterol and therefore,
 may lower the risk of CVD. Eating pulses regularly may also lower CVD risk via favorable
 effects on blood pressure, blood glucose and insulin levels and by satisfying hunger and
 helping manage body weight. Higher legume intakes were associated with lower body mass index
 (BMI), blood pressure, serum total cholesterol (TC) and a lower incidence of T2DM, compared
 with lower legume intakes. In addition to achieving energy balance and improving the overall
 diet, the World Health Organization (WHO) recommends that people increase consumption of
 legumes (pulses), as well as vegetables, wholegrain and nuts to prevent obesity. There does
 not seem to be differential effects of pea-protein as compared to other meat and dairy
 proteins when fed at similar amounts as a pre-load.

 Technically a nut, hempseed typically contains over 30% oil and about 25% protein, with
 considerable amounts of dietary fibre, vitamins and minerals. The two main proteins in
 hempseed are edestin and albumin (high-quality storage proteins are easily digested and
 contain nutritionally significant amounts of all essential amino acids); hempseed has
 exceptionally high levels of the amino acid arginine. Hempseed is an excellent source of
 nutrition and has been used to treat various disorders for thousands of years in traditional
 medicine. Animal studies and only few humans study (on hemp seeds oil) agree as well that
 hempseed has health promoting properties; supported by results from nutritional analyses of
 the seed, oil and seed meal. Being a rich source of polyunsaturated fatty acids and proteins
 which contain all of the essential amino acids in nutritionally sufficient amounts as
 detailed by the FAO/WHO (Food and Agriculture Organization of the United Nations/World Health
 Organization) suggested requirements for infants or children. Hemp proteins have a Protein
 Digestibility Corrected Amino Acid Score (PDCAAS) equal to or greater than certain grains,
 other nuts, and some pulses. Hempseed, in addition to its nutritional value, has demonstrated
 positive health benefits, including the lowering of cholesterol and high blood pressure.

 Buckwheat has attracted increasing attention from food scientists for its potential
 beneficial effects in chronic diseases. In the early 1970's the nutritional properties of
 buckwheat were first demonstrated. The cholesterol lowering effects of buckwheat protein
 extracts in rats with cholesterol-enriched diets has been reported.

 There has been much research conducted on the functionalities and properties of buckwheat
 proteins and other phytochemicals in buckwheat seeds. Among them, the most promising are the
 flavones, flavonoids, phytosterols, fagopyrins, and the thiamin-binding proteins. Buckwheat
 proteins have unique amino acid compositions Experiments, both in animal models and with
 human studies revealed that buckwheat flour can improve T2DM, obesity, hypertension,
 hypercholesterolemia and constipation.

 For the social science element of the project, the proposed study will employ qualitative
 research methods which emphasise the importance of 'lay' knowledge and the subjective
 experience of research participants. Within the field of nutrition research, qualitative
 methods and techniques still remain comparatively rare. This is partly because of the
 privileged status of quantitative research, but also because of the length of time and amount
 of effort that goes into each qualitative study and the small pool of qualified researchers
 in this field .As qualitative research efforts require that researchers seek to understand
 people and the social and cultural contexts in which they live from the participants' points
 of view, this type of knowledge has much to contribute to our understanding of why people eat
 what they do. For the purposes of this study, qualitative methods will enhance our
 understanding of the acceptability, to consumers in Scotland, of foods based on sustainable,
 plant-based protein. It will also tell us much about any barrier that exist to the uptake of
 such foods, from a consumer perspective.

 Planned work for the study

 From a health perspective it is essential to establish the bioavailability and metabolism of
 plant proteins and other dietary constituents in humans. Certain metabolites are absorbed
 early in the gastrointestinal tract, whereas others are transported to the colon where they
 are extensively metabolized by the gut microbiota. In this acute study, we will characterise
 the bioactive metabolites present in the urine and plasma samples obtained. This will inform
 on their availability to the systemic circulation, as well as their excretion profiles. We
 will employ targeted quantitative analysis using both LC-MS and GC-MS which will provide the
 identity of metabolites and their in vivo concentrations. We will focus on metabolites that
 are considered to impact on human health. The principal metabolites analysed will be those
 produced by the phenylpropanoid pathway and products of protein and carbohydrate metabolism.
 These will include derivatives and metabolites of the simple phenols, benzoic acids, phenolic
 acids, phenylacetic acids, phenypropionic acids, phenylpyruvic acids, phenyllactic acids,
 mandellic acids, phenolic dimers, acetophenones, benzaldehydes, cinnamaldehydes, benzyl
 alcohols, cinnamyl alcohols, indoles, isoflavones, coumarins, chalcones, flavanones,
 flavones, flavonols, anthocyanidins. We have also shown that many of these compounds are
 bioactive and exhibit anti-oxidant and anti-inflammatory activity at in vivo concentrations.
 These studies will provide important information on bioavailability and metabolism, but will
 also allow us to correlate the indices with biomarkers of health. In this study we will also
 examine the effect of different protein sources on post-prandial effects, endocrine responses
 (e.g. PYY (peptide YY) levels) and hunger ratings. The energy intake for each volunteer will
 be measured after lunch (ad libitum) on the intervention day. Volunteer views and
 acceptability towards diets rich in plant protein will be assessed. Some components in pulses
 may contribute to weight control by inhibiting intestinal absorption of carbohydrates and
 other energy yielding nutrients such as protein. Correlating the metabolic profiles with
 biomarkers of health will provide information that will be fed back into agricultural
 practice to optimise crop breeding strategies for benefits on human health.

 With regard to the social science component of the project, in-depth semi-structured
 interviews will be used. The term 'semi-structured' reflects the intrinsic flexibility of
 qualitative interview approaches, which often use an aide-memoire or interview guide rather
 than a formal schedule. In addition, the form and content of the research questions is likely
 to vary, in response to each particular interview context. This type of built-in design can
 strengthen qualitative research because of its attentiveness to context.

 All interviews will be digitally recorded and fully transcribed. The qualitative analytical
 techniques applied will look for and examine the themes and patterns of meaning within the
 kinds of data generated by this research mode. The interviews and conversations that take
 place as part of qualitative inquiry will generate responses, perceptions, opinions and so on
 from research participants. These can be understood as constituting a 'discourse' (or set of
 ideas) derived from the focus of the inquiry that lends itself particularly well to thematic
 analysis. Themes are derived by the process known as 'coding' (or labelling) specific chunks
 of text according to the ideas which they seem to represent. This interpretative work
 involves the creation of analytical themes by the researcher and is particularly useful in
 bringing together the 'insider' or 'emic' perspective derived from the research participant's
 knowledge and experience, with the 'outsider' or 'etic' perspective of the researcher and the
 relevant literature. Patterns in the data are thus identified through the intellectual work
 of bringing together components or fragments of ideas, perceptions or experiences which might
 be meaningless when considered in isolation. The researcher engaged in thematic analysis
 seeks to understand how different ideas or components fit together in a meaningful way, in
 order to produce a coherent analytical account.

 The analysis will be validated by referring back to the relevant literature, which may
 support (or counter) inferences drawn by the researcher. The interweaving of research
 findings with extant research literature will help strengthen the interpretive account
 provided by the researcher. Research participants will also be engaged in this process in
 that their feedback on emerging patterns will be sought. This process of member checking will
 be conducted during the course of each interview. Through an analytical framework that
 focuses on discourse and acknowledges the importance of participants' own perspectives and
 experiences, analysis will present a contextualised exploration of the ways in which
 consumers perceive plant-based proteins as alternatives or supplements to meat.
Inclusion Criteria:

 Healthy males and females, non-vegetarians, non-smokers, age 18-65 with BMI 18-35 kg m-2,
 taking no exclusion criteria medication.

 Exclusion Criteria:

 Diabetes Severe gastrointestinal disorders Kidney disease Thromboembolic or coagulation
 disease Hepatic disease Alcohol or any other substance abuse Gout Eating disorders Allergy
 Unregulated thyroid disease Asthma Eczema Hay fever Gluten/wheat intolerance Psychiatric
 disorder resulting in a perceived inability to give informed consent (including severe
 depression, lithium treatment, schizophrenia, severe behavioral disorders)

 Medication exclusion criteria (Confirmed with GP) Orlistat (Xenical) Oral antidiabetics,
 insulin Digoxin, anti-arrhythmics Anti-inflammatories/anti-pyretics Tricyclic
 antidepressants, neuroleptics Antihistamines