The Effect of Regular Consumption of Reformulated Breads on Glycemic Control: A Systematic Review and Meta-Analysis of Randomized Clinical Trials.

Bread is a major source of grain-derived carbohydrates worldwide. High intakes of refined grains, low in dietary fiber and high in glycemic index, are linked with increased risk for type 2 diabetes mellitus (T2DM) and other chronic diseases. Hence, improvements in the composition of bread could influence population health. This systematic review evaluated the effect of regular consumption of reformulated breads on glycemic control among healthy adults, adults at cardiometabolic risk or with manifest T2DM. A literature search was performed using MEDLINE, Embase, Web of Science and the Cochrane Central Register of Controlled Trials. Eligible studies employed a bread intervention (≥2 wk) in adults (healthy, at cardiometabolic risk or manifest T2DM) and reported glycemic outcomes (fasting blood glucose, fasting insulin, HOMA-IR, HbA1c, and postprandial glucose responses). Data were pooled using generic inverse variance with random-effects model and presented as mean difference (MD) or standardized MD between treatments with 95% CIs. Twenty-two studies met the inclusion criteria (n = 1037 participants). Compared with "regular" or comparator bread, consumption of reformulated intervention breads yielded lower fasting blood glucose concentrations (MD: -0.21 mmol/L; 95% CI: -0.38, -0.03; I2 = 88%, moderate certainty of evidence), yet no differences in fasting insulin (MD: -1.59 pmol/L; 95% CI: -5.78, 2.59; I2 = 38%, moderate certainty of evidence), HOMA-IR (MD: -0.09; 95% CI: -0.35, 0.23; I2 = 60%, moderate certainty of evidence), HbA1c (MD: -0.14; 95% CI: -0.39, 0.10; I2 = 56%, very low certainty of evidence), or postprandial glucose response (SMD: -0.46; 95% CI: -1.28, 0.36; I2 = 74%, low certainty of evidence). Subgroup analyses revealed a beneficial effect for fasting blood glucose only among people with T2DM (low certainty of evidence). Our findings suggest a beneficial effect of reformulated breads high in dietary fiber, whole grains, and/or functional ingredients on fasting blood glucose concentrations in adults, primarily among those with T2DM. This trial was registered at PROSPERO as CRD42020205458.

Impact of Daily Consumption of Whole-Grain Quinoa-Enriched Bread on Gut Microbiome in Males.

Diets rich in whole grains are associated with improved health and a lower risk of non-communicable diseases, but the mechanisms through which these health benefits are conveyed are uncertain. One mechanism may be improvements in the gut environment by the delivery of fermentable substrates and associated phytochemicals to the lower gut and modification of the gut microbiome. Quinoa is included in the whole-grain category because of its structural similarities to cereals but the effects of its consumption on the gut microbiome have not been investigated to date. Our aim was to examine the impact of daily quinoa consumption on the gut microbiome in a 4-week randomised cross-over intervention separated by a 4-week wash-out period involving 28 adult males. Participants consumed either a quinoa-enriched wheat-bread roll providing 20 g quinoa flour each day, or a control wheat-only bread roll. Stool samples were collected in sterile collection tubes immediately before and at the end of each intervention period. DNA was then extracted, and the 16S rRNA V4 region of extracted DNA was amplified and sequenced. For both the control and quinoa bread periods, there were no changes at the phyla or genus level between baseline and week 4 (all p > 0.05). Diversity in the microbiome profile was not different from baseline after either intervention arms. The results show that small changes in the type of cereal consumed­substituting 20 g of refined wheat flour with whole-grain quinoa flour­was not able to significantly modulate the gut microbiome. Further studies with higher levels of quinoa or longer exposure periods are needed to ascertain if there is a dose−response effect of quinoa, and if these effects are able to translate into clinical outcomes.

Diet, but not food type, significantly affects micronutrient and toxic metal profiles in urine and/or plasma; a randomized, controlled intervention trial.

BACKGROUND: Observational studies have linked Mediterranean Diets (MedDiets) and organic food consumption with positive health outcomes, which may be explained by higher mineral micronutrient and phenolic intake and lower dietary exposure to toxic compounds. OBJECTIVES: We aimed to determine the effects of diet and food type (organic compared with conventional) on urinary excretion (UE) and/or plasma concentrations of mineral micronutrients, phenolics, and toxic metals. METHODS: Healthy adult participants were randomly allocated to a conventional (n = 14) or an intervention (n = 13) group. During a 2-wk period, the intervention group consumed a MedDiet made entirely from organic foods, whereas the conventional group consumed a MedDiet made from conventional foods. Before and after the intervention period, both groups consumed their habitual Western diets made from conventional foods. The primary outcome was UE and/or plasma concentrations of selected mineral micronutrients, toxic metals, and phenolic markers. In addition, we monitored diets using food diaries. The participants were aware of study group assignment, but the study assessors were not. RESULTS: Changing from a Western Diet to a MedDiet for 2 wk resulted in significant increases in UE of total phenolics and salicylic acid (by 46% and 45%, respectively), the mineral micronutrients Co, I, and Mn (by 211%, 70%, and 102%, respectively), and the toxic metal Ni (by 42%), and plasma Se concentrations (by 14%). However, no significant effects of food type (organic compared with conventional) were detected. Redundancy analysis identified vegetables, coffee, wine, and fruit as positive drivers for UE of phenolic markers and mineral micronutrients, and fish consumption as a positive driver for UE of Cd and Pb. CONCLUSIONS: Although small effects of food type cannot be ruled out, our study suggests that only changing to a MedDiet with higher fruit and vegetable, and lower meat, consumption results in a large increase in phenolic and mineral micronutrient intakes. This trial was registered at clinicaltrials.gov as NCT03254537.

Double-blind controlled dietary cross-over intervention with differentially fertilised intact lettuce leaves shows acute reduction in blood pressure in young adults, associated with faster uptake of nitrate than of phenolics.

PURPOSE: To compare acute effects on blood pressure (BP) of ingestion of visually similar lettuce with controlled high and low content of either nitrate or phenolic compounds. METHODS: In a randomised cross-over design, 19 healthy participants (22-31 years) received 50 g of lettuce containing either 530 mg (8.4 mmol) nitrate + 11 mg (0.03 mmol) phenolic compounds (HNLP); or 3 mg nitrate (0.05 mmol) + 77 mg (0.2 mmol) phenolic compounds (LNHP), obtained by differential fertilisation. Ambulatory BP was recorded along with plasma, salivary and urinary nitrate and nitrite and plasma concentrations of cyclic guanosine monophosphate (cGMP), phenolic metabolites, Trolox equivalent antioxidant capacity (TEAC) and ferric reducing antioxidant power (FRAP). RESULTS: Compared with LNHP, 3 h post ingestion of HNLP, plasma nitrate increased 0.31 ± (95%CI) 0.12 mM (+ 240%), and salivary nitrate 5.5 ± 1.4 mM (+ 910%); accumulated urinary nitrate excretion increased 188 ± 72 mg (+ 296%) (all P < 0.001). Systolic BP was reduced 4.9 ± 4.2 mmHg (P = 0.031) between 3 and 6 h after ingestion of HNLP compared with LNHP; systolic BP differences were negatively correlated (P = 0.004) with differences in saliva nitrate concentrations. LNHP increased plasma phenolics at 6 h, predominantly 3'-methoxycinnamic acid-4'-glucuronide (ferulic acid-4'-glucuronide), 116%, 204 ± 138 nM more than HNLP (P = 0.001); increased cGMP 14% (P = 0.019); and reduced FRAP 3.1% (P = 0.009). CONCLUSION: The acute BP difference within 6 h of consumption matched the plasma/saliva nitrate peak, not the slower changes of plasma phenolics. This is the first double-blind controlled dietary intervention demonstrating differential effects on human physiology by consumption of an intact plant food, where compositional differences were obtained by controlling growing conditions, indicating potential opportunities for health claims relating to precision/vertical farming. CLINICAL TRIAL REGISTRATION: The trial was retrospectively registered on ClinicalTrials.gov, with identifier NCT02701959, on March 8, 2016.

Diet and food type affect urinary pesticide residue excretion profiles in healthy individuals: results of a randomized controlled dietary intervention trial.

BACKGROUND: Observational studies have linked pesticide exposure to various diseases, whereas organic food consumption has been associated with positive health outcomes. Organic farming standards prohibit the use of most pesticides, and organic food consumption may therefore reduce pesticide exposure. OBJECTIVES: To determine the effects of diet (Western compared with Mediterranean) and food type (conventional compared with organic) and sex on urinary pesticide residue excretion (UPRE), as well as associations between specific diet components and UPRE. METHODS: In this 2-wk, randomized dietary intervention trial, healthy adults were randomly allocated to an intervention (n = 13) or conventional (n = 14) group. Whereas participants in the intervention group consumed a Mediterranean diet (MedDiet) made entirely from organic foods, the conventional group consumed a MedDiet made entirely from conventional foods. Both groups consumed habitual Western diets made from conventional foods before and after the 2-wk intervention period. The primary outcome was UPRE. In addition, we assessed diet composition and pesticide residue profiles in foods eaten. Participants were aware of group assignment, but the study assessors were not. RESULTS: During the intervention period, total UPRE was 91% lower with organic (mean 17 µg/d; 95% CI: 15, 19) than with conventional (mean 180 µg/d; 95% CI: 153, 208) food consumption (P < 0.0001). In the conventional group, switching from the habitual Western diet to the MedDiet increased insecticide excretion from 7 to 25 µg/d (P < 0.0001), organophosphate excretion from 5 to 19 µg/d (P < 0.0001), and pyrethroid residue excretion from 2.0 to 4.5 µg/d (P < 0.0001). Small but significant effects of sex were detected for chlormequat, herbicide, and total pesticide residue excretion. CONCLUSIONS: Changing from a habitual Western diet to a MedDiet was associated with increased insecticide, organophosphate, and pyrethroid exposure, whereas organic food consumption reduced exposure to all groups of synthetic chemical pesticides. This may explain the positive health outcomes linked to organic food consumption in observational studies. This trial was registered at www.clinicaltrials.gov as NCT03254537.

Aligning nutrient profiling with dietary guidelines: modifying the Nutri-Score algorithm to include whole grains.

PURPOSE: Whole grains, generally recognised as healthy choices, are not included in most nutrient profiling systems. We tested modifications to the Nutri-Score algorithm to determine whether including whole grains would provide an improved measure of food, and overall diet quality. METHODS: The whole-grain content of food, with a minimum cut-point of 25%, was added to the algorithm, following similar methods used to score other health-promoting components such as fibre. We applied and compared the original and the modified Nutri-Score to food composition and dietary intake data from Australia, France, the United Kingdom, and the United States. RESULTS: At the food level, correlations between whole-grain content and food nutritional score were strengthened using the modified algorithm in Australian data, but less so for the other countries. Improvements were greater in grain-specific food groups. The largest shift in Nutri-Score class was from B to A (best score). At the dietary intake level, whole-diet nutritional scores for individuals were calculated and compared against population-specific diet-quality scores. With modifications, correlations with diet-quality scores were improved slightly, suggesting that the modified score better aligns with national dietary guidelines. An inverse linear relationship between whole-diet nutritional score and whole-grain intake was evident, particularly with modifications (lower whole-diet nutritional score indicative of better diet quality). CONCLUSION: Including a whole-grain component in the Nutri-Score algorithm is justified to align with dietary guidelines and better reflect whole grain as a contributor to improved dietary quality. Further research is required to test alternative algorithms and potentially other nutrient profiling systems.

Opportunities for diet quality improvement: the potential role of staple grain foods.

OBJECTIVE: Given the high disease burden associated with the low intake of whole grains, modelling studies that estimate the impact of dietary strategies to increase more healthful grain foods consumption are essential to inform evidence-based and culturally specific policies. The current study investigated the potential nutritional impact of replacing staple grain foods with more healthful options. DESIGN: Based on the 2015 Health Survey of São Paulo, a cross-sectional, population-based study, we modelled the substitution of white rice and white bread with brown rice and whole-wheat bread. Outcomes included changes in more healthful grain foods, energy and nutrient intakes. SETTING: Urban area of São Paulo, Brazil. PARTICIPANTS: Participants aged over 12 years who completed a semi-structured questionnaire and one 24-h recall (n 1741). RESULTS: The substitution of all white rice and white bread with brown rice and whole-wheat bread, respectively, would result in more than 5 % increases in Zn (+9·1 %), Ca (+9·3 %), vitamin E (+18·8 %), dietary fibre (+27·0 %) and Mg (+52·9 %) intake, while more than a 5 % decrease would be seen for total carbohydrate (-6·1 %), folate (-6·6 %), available carbohydrate (-8·5 %), Fe (-8·6 %), vitamin B6 (-12·5 %), vitamin B2 (-17·4 %), and vitamin B1 (-20·7 %). A substantial increase in the amount of more healthful grain foods consumed would be seen (10 g/d to 220 g/d, or from 4 % to 69 % of total grain intake). CONCLUSIONS: Replacing white rice and white bread with their whole-grain versions has the potential to improve diet quality, suggesting they are prime targets for policy actions aiming at increasing intake of more healthful grain foods.

Health benefits of whole grain: effects on dietary carbohydrate quality, the gut microbiome, and consequences of processing.

Grains are important sources of carbohydrates in global dietary patterns. The majority of these carbohydrates, especially in refined-grain products, are digestible. Most carbohydrate digestion takes place in the small intestine where monosaccharides (predominantly glucose) are absorbed, delivering energy to the body. However, a considerable part of the carbohydrates, especially in whole grains, is indigestible dietary fibers. These impact gut motility and transit and are useful substrates for the gut microbiota affecting its composition and quality. For the most part, the profile of digestible and indigestible carbohydrates and their complexity determine the nutritional quality of carbohydrates. Whole grains are more complex than refined grains and are promoted as part of a healthy and sustainable diet mainly because the contribution of indigestible carbohydrates, and their co-passenger nutrients, is significantly higher. Higher consumption of whole grain is recommended because it is associated with lower incidence of, and mortality from, CVD, type 2 diabetes, and some cancers. This may be due in part to effects on the gut microbiota. Although processing of cereals during milling and food manufacturing is necessary to make them edible, it also offers the opportunity to still further improve the nutritional quality of whole-grain flours and foods made from them. Changing the composition and availability of grain carbohydrates and phytochemicals during processing may positively affect the gut microbiota and improve health.

Perspective: Why Whole Grains Should Be Incorporated into Nutrient-Profile Models to Better Capture Nutrient Density.

Healthy eating patterns, as described by dietary guidelines, typically favor whole grains, low-fat dairy, vegetables, fruit, legumes, and nuts and seeds. Nutrient-profiling (NP) models capture nutrient density of individual foods and can inform healthier food choices. Although whole grains are prominently featured in most dietary guidelines, they are not included in most NP models. Healthy foods, as identified by most NP models, are those that contain limited amounts of energy, saturated fat, total or added sugar, and sodium. As global dietary guidance turns to foods and food groups as opposed to individual nutrients, future nutrient-density metrics may need to do the same. Potential methods to incorporate whole grains into the overall concept of nutrient density and into selected NP models are outlined in this review. Incorporating whole grains into the Nutri-Score, Health Star Rating, or the Nutrient Rich Food index will require further analyses of dietary nutrient density in relation to health outcomes across diverse population subgroups. We present the rationale for how the inclusion of whole grains in NP models can assist in the implementation of dietary guidance.

Consensus, Global Definitions of Whole Grain as a Food Ingredient and of Whole-Grain Foods Presented on Behalf of the Whole Grain Initiative.

Proposed global definitions of whole grain as an ingredient and whole grain food are presented by the authors on behalf of the Whole Grain Initiative. Whole grains are an important pillar of healthy and sustainable diets. Internationally accepted credible definitions of whole grains as food ingredients and whole-grain foods are necessary to ensure that all global stakeholders have shared standards, and that consumers find them clear, credible, and useful. Based on widely accepted, existing definitions and new developments, the Definitions Working Group of the global Whole Grain Initiative, with experts from academia, government agencies and industry, developed definitions for global application. The key statements of the definition documents are as follows: "Whole grains shall consist of the intact, ground, cracked, flaked or otherwise processed kernel after the removal of inedible parts such as the hull and husk; all anatomical components, including the endosperm, germ, and bran must be present in the same relative proportions as in the intact kernel" and "A whole-grain food shall contain at least 50% whole-grain ingredients based on dry weight. Foods containing 25-50% whole-grain ingredients based on dry weight, may make a front-of-pack claim on the presence of whole grain but cannot be designated 'whole grain' in the product name". The definition documents have been ratified by the leading international scientific associations in this area. We urge that these consensus Whole Grain Initiative definitions be adopted as the basis for definitions used by national regulatory authorities and for health promotion organisations worldwide to use in nutrition education and food labelling.

The effect of seaweed enriched bread on carbohydrate digestion and the release of glucose from food.

Two seaweeds; Ascophyllum nodosum and Fucus vesiculosus, were incorporated into bread at 0.5 and 2% and their effect on blood glucose in vivo and carbohydrate digestion in vitro were studied. In the five way randomised placebo controlled double blind pilot trial (n = 10) each volunteer consumed 100 g of available carbohydrate (from bread) and their blood glucose was measured over two hours. The breads were tested in a human digestion model and compared against control bread and control bread with the equivalent amount of seaweed. In the pilot human study the enriched breads did not cause any significant reductions in iAUC of blood glucose with average reductions of 0.1 ± 44.4%, 8.2 ± 19.3%, 1.0 ± 54.3% and 2.7 ± 31.9% for 0.5% F.vesiculosus, 0.5% A.nodosum, 2% F.vesiculosus, and 2% A.nodosum respectively. However, seaweed added alongside the control bread in vitro significantly reduced the level of carbohydrate digestion compared to the control bread. F.vesiculosus or A.nodosum can reduce carbohydrate digestion, however baking into bread reduces the effect.

PROTOCOL: The association between whole-grain dietary intake and noncommunicable diseases: A systematic review and meta-analysis.

Our primary research questions are: (1) What is the association between whole grains (WG) intake and the prevalence of NCDs (i.e., type 2 diabetes, cardiovascular disease, obesity, cancer, mortality) and their biomarkers? (2) Which biomarker(s) has/have the greatest association with WG intake when combining multiple biomarkers together in the same analysis? Our secondary research question is: (3) Are there dose-response relationships between WG intake and biomarkers and prevalence of NCDs which could help inform a universal recommendation for WG intake?

Effects of inorganic nitrate and vitamin C co-supplementation on blood pressure and vascular function in younger and older healthy adults: A randomised double-blind crossover trial.

BACKGROUND: Vitamin C and inorganic nitrate have been linked to enhanced nitric oxide (NO) production and reduced oxidative stress. Vitamin C may also enhance the conversion of nitrite into NO. AIMS: We investigated the potential acute effects of vitamin C and inorganic nitrate co-supplementation on blood pressure (BP) and peripheral vascular function. The secondary aim was to investigate whether age modified the effects of vitamin C and inorganic nitrate on these vascular outcomes. METHODS: Ten younger (age 18-40 y) and ten older (age 55-70 y) healthy participants were enrolled in a randomised double-blind crossover clinical trial. Participants ingested a solution of potassium nitrate (7 mg/kg body weight) and/or vitamin C (20 mg/kg body weight) or their placebos. Acute changes in resting BP and vascular function (post-occlusion reactive hyperemia [PORH], peripheral pulse wave velocity [PWV]) were monitored over a 3-h period. RESULTS: Vitamin C supplementation reduced PWV significantly (vitamin C: -0.70 ± 0.31 m/s; vitamin C placebo: +0.43 ± 0.30 m/s; P = 0.007). There were significant interactions between age and vitamin C for systolic, diastolic, and mean arterial BP (P = 0.02, P = 0.03, P = 0.02, respectively), with systolic, diastolic and mean BP decreasing in older participants and diastolic BP increasing in younger participants following vitamin C administration. Nitrate supplementation did not influence BP (systolic: P = 0.81; diastolic: P = 0.24; mean BP: P = 0.87) or vascular function (PORH: P = 0.05; PWV: P = 0.44) significantly in both younger and older participants. However, combined supplementation with nitrate and vitamin C reduced mean arterial BP (-2.6 mmHg, P = 0.03) and decreased PWV in older participants (PWV: -2.0 m/s, P = 0.02). CONCLUSIONS: The co-administration of a single dose of inorganic nitrate and vitamin C lowered diastolic BP and improved PVW in older participants. Vitamin C supplementation improved PWV in both age groups but decreased systolic and mean BP in older participants only. CLINICAL TRIAL REGISTRATION: Current Controlled Trials (ISRCTN98942199).

A Vitamin D, Calcium and Leucine-Enriched Whey Protein Nutritional Supplement Improves Measures of Bone Health in Sarcopenic Non-Malnourished Older Adults: The PROVIDE Study.

Alterations in musculoskeletal health with advanced age contribute to sarcopenia and decline in bone mineral density (BMD) and bone strength. This decline may be modifiable via dietary supplementation. To test the hypothesis that a specific oral nutritional supplement can result in improvements in measures of bone health. Participants (n 380) were participants of the PROVIDE study, a 13-week, multicenter, randomized, controlled, double-blind, 2 parallel-group study among non-malnourished older participants (≥ 65 years) with sarcopenia [determined by Short Physical Performance Battery (SPPB; 0-12) scores between 4 and 9, and a low skeletal muscle mass index (SMI; skeletal muscle mass/BW × 100) ≤ 37% in men and ≤ 28% in women using bioelectric impedance analysis] Supplementation of a vitamin D, calcium and leucine-enriched whey protein drink that comprises a full range of micronutrients (active; 2/day) was compared with an iso-caloric control. Serum 25-hydroxyvitamin D [25(OH)D], parathyroid hormone (PTH), biochemical markers of bone formation (osteocalcin; OC, procollagen type 1 amino-terminal propeptide; P1NP) and resorption (carboxy-terminal collagen crosslinks; CTX), insulin like growth factor 1 (IGF-1) and total-body BMD were analysed pre- and post-intervention. Serum 25(OH)D concentrations increased from 51.1 ± 22.9 nmol/L (mean ± SD) to 78.9 ± 21.1 nmol/L in the active group (p < 0.001 vs. control). Serum PTH showed a significant treatment difference (p < 0.001) with a decline in the active group, and increase in the control group. Serum IGF-1 increased in the active group (p < 0.001 vs. control). Serum CTX showed a greater decline in the active group (p = 0.001 vs. control). There were no significant differences in serum OC or P1NP between groups during the intervention. Total body BMD showed a small (0.02 g/cm2; ~ 2%) but significant increase in the active group after supplementation (p = 0.033 vs. control). Consuming a vitamin D, calcium and leucine-enriched whey protein supplement for 13 weeks improved 25(OH)D, suppressed PTH and had small but positive effects on BMD, indicative of improved bone health, in sarcopenic non-malnourished older adults.

Acceptability of alginate enriched bread and its effect on fat digestion in humans.

Lifestyle interventions and physical activity remain the cornerstone of obesity management, as pharmacological therapies (orlistat) are associated with gastrointestinal (GI) side effects. Combining orlistat with fibers can reduce side effects, improving compliance. Therefore, a fiber that inhibits lipase without side effects could help treat obesity. The aims of the present work were to assess whether alginate enriched bread could inhibit fat digestion, and assess the acceptability of alginate bread and its effect on GI wellbeing. A double-blind, randomised, controlled cross-over pilot study (NCT03350958) assessed the impact of an alginate bread meal on; lipid content in ileal effluent and circulating triacylglycerol levels. This was compared against the same meal with non-enriched (control) bread. GI wellbeing and acceptability of alginate bread was compared to control bread through daily wellbeing questionnaires and food diaries (NCT03477981). Control bread followed by alginate bread were consumed for two weeks respectively. Consumption of alginate bread reduced circulating triacylglycerol compared to control (2% reduction in AUC) and significantly increased lipid content in ileal effluent (3.8 g ±â€¯1.6 after 210 min). There were no significant changes to GI wellbeing when comparing alginate bread to control bread. A significant increase in the feeling of fullness occurred with alginate bread compared to baseline and the first week of control bread consumption. This study showed that sustained consumption of alginate enriched bread does not alter GI wellbeing and can decrease lipolysis, increasing lipid leaving the small intestine. Further studies are required to demonstrate that reduced fat digestion through the action of alginate can reduce fat mass or body weight.

Elevated Total Homocysteine in All Participants and Plasma Vitamin B12 Concentrations in Women Are Associated With All-Cause and Cardiovascular Mortality in the Very Old: The Newcastle 85+ Study.

Background: Folate and vitamin B12 are keys to the correct functioning of one-carbon (1-C) metabolism. The current evidence on associations between 1-C metabolism biomarkers and mortality is inconclusive and generally based on younger or institutionalized populations. This study aimed to determine the associations between biomarkers of 1-C metabolism and all-cause and cardiovascular (CVD) mortality in the very old. Methods: The Newcastle 85+ Study is a prospective longitudinal study of participants aged 85 at recruitment living in Northeast England. Baseline red blood cell folate (RBC folate), plasma vitamin B12, and total homocysteine (tHcy) concentrations were available for 752-766 participants. Associations between biomarkers of 1-C metabolism and all-cause and CVD mortality for up to 9 years were assessed by Cox proportional hazard models and confirmed by restricted cubic splines. Results: Participants with higher tHcy concentrations had higher risk of death from any cause (hazard ratio [HR] [×10 µmol/L]: 1.24, 95% confidence interval [CI]: 1.10-1.41) and cardiovascular diseases (HR [×10 µmol/L]: 1.23, 95% CI: 1.04-1.45) than those with lower concentrations; and women with higher plasma vitamin B12 concentrations had increased risk of all-cause and cardiovascular mortality (HR [×100 pmol/L]: 1.10, 95% CI: 1.04-1.16) after adjustment for key sociodemographic, lifestyle, and health confounders. Conclusion: Higher concentrations of tHcy in all participants and plasma vitamin B12 in women were associated with increased risk of all-cause and CVD mortality in the very old. This confirms findings for tHcy in younger populations but the adverse relationships between elevated plasma vitamin B12 concentrations and mortality in this setting are novel and require further investigation.

Manipulation of Contents of Nitrate, Phenolic Acids, Chlorophylls, and Carotenoids in Lettuce (Lactuca sativa L.) via Contrasting Responses to Nitrogen Fertilizer When Grown in a Controlled Environment.

This study aimed to use different nitrogen fertilizer regimes to produce Butterhead lettuce with such large differences in nitrate content that they could be used as treatment and placebo to study the effect of inorganic nitrate on human health. Plants were grown under controlled conditions at 27/23 °C day/night with a relatively low photosynthetically active radiation (PAR) of 150 µmol m-2 s-1 for 14 h day-1 and nitrogen supplies ranging from 26 to 154 ppm of N as ammonium nitrate in the fertigation solution. This resulted in contrasting high (∼1078 mg nitrate 100 g-1 FW) or low (∼6 mg 100 g-1) nitrate contents in the leaves. Contents of carotenoids and chlorophylls in fresh weight did not differ significantly between the highest and the lowest N-supply levels. However, increased nitrogen supply reduced contents of phenolic compounds from 154 to 22 mg 100 g-1 FW, dry matter content from 8.9% to 4.6%, and fresh weight per plant from 108.52 to 47.57 g/plant FW (all P < 0.001). Thus, while fertilizer treatments can provide lettuce with substantially different nitrate contents, maintaining similar pigment contents (color), they also strongly influence the contents of phenolic acids and flavones.