Exposure to undernutrition during fetal life has been proposed as an underlying cause of adult hypertension. Epidemiological studies demonstrating relationships between low birth weight and later CVD are supported by animal experiments indicating that manipulations of the maternal diet in pregnancy exert programming effects upon blood pressure control. Pregnant female Wistar rats were fed a control diet (n 13) or a low-protein diet (n 12) throughout pregnancy. At delivery all animals were fed the same standard laboratory chow diet. Analysis of nephron number in kidneys obtained from 4-week-old offspring showed that this was significantly (P<0·05) reduced in animals exposed to maternal protein restriction. At this age rats exposed to low-protein diets in utero had systolic blood pressures that were significantly greater than those of control animals (+23 mmHg, P<0·05). Administration of ascending doses of angiotensin II (1–40 ng/kg body weight intravenously) to 10-week-old anaesthetised female rats showed that the pressor response to the peptide was greater and more prolonged in animals exposed to low-protein diets in utero. Renal expression of mRNA for the angiotensin II type 1A receptor was similar in the two groups of rats, but low-protein-exposed animals had significantly lower renal expression of the type 2 receptor (P=0·023). These results suggest that maternal nutritional status programmes expression of the renal angiotensin II type 2 receptor. This may play a key role in the impairment of renal development and the elevation of blood pressure noted in rats exposed to intra-uterine protein restriction.

A low HDL-cholesterol concentration is an independent risk factor for CVD. Studies have suggested that flavonoid consumption may be cardioprotective, and a favourable impact on circulating HDL-cholesterol concentrations has been suggested to partially explain this association. The aim of the present study was to determine the effect of consuming increasing daily doses of low-calorie cranberry juice cocktail (CJC) on the plasma lipid profile of abdominally obese men. For that purpose, thirty men (mean age 51 (SD 10) years) consumed increasing doses of CJC during three successive periods of 4 weeks (125ml/d, 250ml/d, 500ml/d). Before the study and after each phase, we measured changes in physical and metabolic variables. We noted a significant increase in plasma HDL-cholesterol concentration after the consumption of 250ml CJC/d (+8·6±14·0% v. 0ml CJC/d; P<0·01), an effect that plateaued during the last phase of the study (500ml CJC/d: +8·1±10·0% v. 0ml CJC/d; P<0·0001). Multivariate analyses revealed that changes in plasma apo A-I (IR2=48%, P<0·0001) and triacylglycerol (R2=16%, P<0·005) concentrations were the only variables significantly contributing to the variation in plasma HDL-cholesterol concentration noted in response to the intervention. No variation was observed in total as well as in LDL and VLDL cholesterol. The present results show that daily CJC consumption is associated with an increase in plasma HDL-cholesterol concentrations in abdominally obese men. We hypothesise that polyphenolic compounds from cranberries may be responsible for this effect, supporting the notion that the consumption of flavonoid-rich foods can be cardioprotective.

The consumption of cocoa and dark chocolate is associated with a lower risk of CVD, and improvements in endothelial function may mediate this relationship. Less is known about the effects of cocoa/chocolate on the augmentation index (AI), a measure of vascular stiffness and vascular tone in the peripheral arterioles. We enrolled thirty middle-aged, overweight adults in a randomised, placebo-controlled, 4-week, cross-over study. During the active treatment (cocoa) period, the participants consumed 37 g/d of dark chocolate and a sugar-free cocoa beverage (total cocoa = 22 g/d, total flavanols (TF) = 814 mg/d). Colour-matched controls included a low-flavanol chocolate bar and a cocoa-free beverage with no added sugar (TF = 3 mg/d). Treatments were matched for total fat, saturated fat, carbohydrates and protein. The cocoa treatment significantly increased the basal diameter and peak diameter of the brachial artery by 6 % (+2 mm) and basal blood flow volume by 22 %. Substantial decreases in the AI, a measure of arterial stiffness, were observed in only women. Flow-mediated dilation and the reactive hyperaemia index remained unchanged. The consumption of cocoa had no effect on fasting blood measures, while the control treatment increased fasting insulin concentration and insulin resistance (P= 0·01). Fasting blood pressure (BP) remained unchanged, although the acute consumption of cocoa increased resting BP by 4 mmHg. In summary, the high-flavanol cocoa and dark chocolate treatment was associated with enhanced vasodilation in both conduit and resistance arteries and was accompanied by significant reductions in arterial stiffness in women.

The tolerance of low-digestible carbohydrates (LDCs) may be measured as the potential to cause abdominal symptoms and laxation. Tolerance of any one LDC is determined by its concentration in the food product eaten, the amount of the food product eaten, the frequency of eating the food and the consumption of other foods (increasing tolerance) and water (decreasing tolerance). Added to these, individuals vary considerably in their response to low-digestible carbohydrates in the reporting of gastrointestinal symptoms. A precise maximum no-response dose is sometimes difficult to obtain because some dose–response curves are distinctly sigmoidal. Food regulators hoping to set a trigger level at which laxation may occur have been unable to take account of all these factors because the necessary information matrices are not available for any one LDC. Nevertheless analysis of the data shows consistent trends and for circumstances when food is consumed throughout the day it now seems feasible to assign specific tolerances to specific low-digestible carbohydrates, especially the polyols for which most is known. The method by which the no-effect dose or laxative threshold is expressed is critical to its application to individual foods.

This meta-analysis examined the effect of probiotics on glucose and glycaemic factors in diabetes and its associated risk factors. All randomised-controlled trials published in English in multiple databases from January 2000 to June 2015 were systematically searched. Only studies that addressed glucose- and glycaemic-related factors as outcome variables were included. The main outcomes of interest in trials were mean changes in glucose, HbA1c, insulin and homoeostasis model assessment-estimated insulin resistance (HOMA-IR). Using the Physiotherapy Evidence Database (PEDro) scale to assess the quality of studies, a total of eleven studies with 614 subjects were included. The pooled mean difference and effect size with a 95 % CI were extracted using a random-effect model. It was found that there are statistically significant pooled mean differences between the probiotics and the placebo-controlled groups on the reduction of glucose (−0·52 mmol/l, 95 % CI −0·92, −0·11 mmol/l; P=0·01) and HbA1c (−0·32 %, 95 % CI −0·57, −0·07 %; P=0·01). There was no statistically significant pooled mean difference between the probiotics and the placebo-controlled groups on the reduction of insulin (−0·48 µIU/ml, 95 % CI −1·34, 0·38 µIU/ml; P=0·27) and HOMA-IR (pooled effect of –0·44, 95 % CI −1·57, 0·70; P=0·45). Meta-regression analysis identified that probiotics had significant effects on reduction of glucose, HbA1c, insulin and HOMA-IR in participants with diabetes, but not in participants with other risk factors. The present meta-analysis suggested that probiotics may be used as an important dietary supplement in reducing the glucose metabolic factors associated with diabetes.

Few studies have explored the relationship between dietary patterns and the risk of gestational diabetes mellitus (GDM). Evidence from non-Western areas is particularly lacking. In the present study, we aimed to examine the associations between dietary patterns and the risk of GDM in a Chinese population. A total of 3063 pregnant Chinese women from an ongoing prospective cohort study were included. Data on dietary intake were collected using a FFQ at 24–27 weeks of gestation. GDM was diagnosed using a 75 g, 2 h oral glucose tolerance test. Dietary patterns were determined by principal components factor analysis. A log-binomial regression model was used to examine the associations between dietary pattern and the risk of GDM. The analysis identified four dietary patterns: vegetable pattern; protein-rich pattern; prudent pattern; sweets and seafood pattern. Multivariate analysis showed that the highest tertile of the vegetable pattern was associated with a decreased risk of GDM (relative risk (RR) 0·79, 95 % CI 0·64, 0·97), compared with the lowest tertile, whereas the highest tertile of the sweets and seafood pattern was associated with an increased risk of GDM (RR 1·23, 95 % CI 1·02, 1·49). No significant association was found for either the protein-rich or the prudent pattern. The protective effect of a high vegetable pattern score was more evident among women who had a family history of diabetes (P for interaction = 0·022). These findings suggest that the vegetable pattern was associated with a decreased risk of GDM, while the sweets and seafood pattern was associated with an increased risk of GDM. These findings may be useful in dietary counselling during pregnancy.

SCFA provide energy to the host and influence lipid and glucose metabolism, suggesting that they may have an impact on the occurrence of metabolic risk factors. The aim of the present study was to determine the concentration of SCFA in faeces of lean and obese individuals and to analyse whether associations between faecal SCFA and metabolic syndrome parameters are present. Lean (n 20) and obese (n 20) women of similar age (28·5 (sd 7·6) v. 30·7 (sd 6·5) years, P= 0·33) participated in the study. Anthropometric measurements, body composition, blood pressure and biochemical parameters were assessed. SCFA were extracted from faeces and quantified by GC. Blood pressure and blood glucose, although within the normal limits, were higher in the obese group compared to lean subjects (P< 0·05). Lower HDL concentration and higher insulin and homeostasis model assessment (HOMA) index were observed in the obese than in the lean group (P< 0·05). The median values of SCFA (% w/w) from the lean and obese groups were butyric (0·021 v. 0·044, P= 0·024), propionic (0·021 v. 0·051, P= 0·007) and acetic (0·03 v. 0·061, P= 0·01). SCFA correlated positively with metabolic syndrome risk factors such as adiposity, waist circumference and HOMA index (P< 0·05), and inversely with HDL (P< 0·05). Our results suggest that the higher faecal concentration of SCFA is associated with metabolic risk factors and thus may influence metabolic homeostasis.

The effects of four prebiotics (fructo-oligosaccharide, Bio-MOS®, transgalacto-oligosaccharide and GroBiotic®-A) on digestive enzymes and intestinal morphology were studied in juvenile hybrid striped bass (Morone chrysops×M. saxatilis) and red drum (Sciaenops ocellatus) using two separate 8-week feeding trials. Red drum were fed experimental diets with the four prebiotics each individually supplemented at 1 % and hybrid striped bass were fed diets supplemented with GroBiotic®-A at 1 and 2 %. Both trials were conducted with each diet fed to apparent satiation twice per d to three replicate groups of fifteen juvenile fish. For histomorphological analysis, gastrointestinal tract (GIT) samples from three randomly selected fish per tank were taken at 4 and 8 weeks for hybrid striped bass and at 8 weeks for red drum. For both trials, GIT samples from two randomly selected fish per tank were taken at 4 and 8 weeks and analysed for pepsin, trypsin, chymotrypsin, aminopeptidase, α-amylase, lipase, and both acid and alkaline phosphatase activities. The results of the histological evaluation indicated that the inclusion of prebiotics was adequate to elicit structural changes in the GIT of both species. On the other hand, no significant changes in the enzyme activities were detected at week 8 in both species. However, there was a transient effect of Bio-MOS® supplementation on the activities of aminopeptidase, α-amylase and alkaline phosphatase at week 4 in red drum only. Thus, previously observed improvements in nutrient digestibility by these fish in response to prebiotic supplementation appear to be mostly related to changes in GIT structure as opposed to the enhancement of digestive enzyme activity.

The bulking index (i.e. the increase in faecal fresh weight in gram per gram indigestible carbohydrate ingested) with oligofructose and inulin is similar to that produced with other easily fermented fibres such as pectins and gums. Most studies in man have been performed at a level of 15 g/d and more investigations on lower intakes are needed to appoint the least intake for an effect. Concerning short-chain fatty acids (SCFA) most studies have been using oligofructose and points at an increased butyric acid formation in the caecum of rats. In one study on rats with inulin high caecal proportions of propionic acid were obtained. As inulin has a higher molecular weight than oligofructose it might be speculated if this could be a reason to the different SCFA-profile formed. No effects on faecal concentrations of SCFA in humans have been revealed with inulin and oligofructose, which neither is expected as most of the SCFA formed during the fermentation already has been absorbed or utilized by the colonic mucosa.

Intervention studies have demonstrated that saturated fat increases total and LDL-cholesterol concentrations, and it is therefore recommended that the intake of high-fat dairy products be limited. However, observational studies have found an inverse relationship between the intake of dairy products and incidence of CVD. We aimed to study whether the Ca content of dairy products influences the effect of dairy fat on the lipid profile. The study had a randomised cross-over design. Subjects (n 9) were randomised to one of the sequence of four isoenergetic 10 d diets: low Ca and low fat (LC/LF: approximately 700 mg Ca/d, 25 % of energy (fat); high Ca and LF (HC/LF: approximately 2800 mg Ca/d, 25 % of energy fat); LC and high fat (LC/HF: approximately 700 mg Ca/d, 49 E% fat); or HC and HF (approximately 2800 mg Ca/d, 49 E% fat). Blood variables were measured before and after each diet period, and faeces and urine were collected at the end of each diet period. A two-way ANOVA was used to examine the effect of Ca and fat intake. Independent of Ca intake, the HF diet increased the concentrations of total (9 %; P < 0·0001), LDL (14 %; P < 0·0001)- and HDL (13 %; P = 0·0002)-cholesterol compared with the LF diet. However, independent of fat intake, the HC diet decreased the concentrations of total (4 %; P = 0·0051) and LDL-cholesterol (10 %; P < 0·0001) but not HDL-cholesterol compared with the LC diet. In addition, total:HDL-cholesterol was decreased (5 %; P = 0·0299), and HDL:LDL was increased (12 %; P = 0·0097) by the HC diet compared with the LC diet. Faecal fat excretion was increased by both the HC (P < 0·0001) and HF (P = 0·0052) diets. In conclusion, we observed that dairy Ca seems to partly counteract the raising effect of dairy fat on total and LDL-cholesterol, without reducing HDL-cholesterol.

The aim of this study was to evaluate the clinical utility of appetite sensations to characterize individual overall energy intake. A group of men (n 28) and women (n 23) was recruited to record their ‘desire to eat’, ‘hunger’, ‘fullness’ and ‘prospective food consumption’ (PFC) on visual analogue scales before a standardized meal test, immediately after and every 10 min for a period of 1 h after the meal. The 1 h post-meal area under the curve (1 h AUC) and the satiety quotient (SQ) were calculated for all appetite sensations. In a second visit, all participants were invited to eat three meals in order to measure total energy intake (TEI) and food preferences. Metabolic rate (MR) was also assessed to derive daily relative energy intake (REI) by subtracting this variable from TEI (TEI−MR=REI). The Three-Factor Eating Questionnaire scores were also calculated for all participants. One h AUC for fullness was the appetite sensation most strongly associated with TEI and REI (r−0·42, P≤0·003 and r−0·32, P≤0·05, respectively). SQ for fullness was the only predictor of TEI and REI (r−0·42, P≤0·0003 and r−0·30, P≤0·05, respectively). Restraint, disinhibition and hunger scores were not associated with appetite sensation variables. These results suggest that the fullness dimension seems to be a useful appetite sensation to predict long-term TEI and REI. Thus, assessment of appetite sensation such as fullness in response to a fixed load may be useful to evaluate individual overall energy intake.

Human milk (HM) is rich in oligosaccharides (HMO) that exert prebiotic and anti-infective activities. HM feeding reduces the incidence of rotavirus (RV) infection in infants. Herein, the anti-RV activity of oligosaccharides was tested in an established in vitro system for assessing cellular binding and viral infectivity/replication, and also tested in a newly developed, acute RV infection, in situ piglet model. For the in vitro work, crude HMO isolated from pooled HM, neutral HMO (lacto-N-neotetraose, LNnT; 2′-fucosyllactose) and acidic HMO (aHMO, 3′-sialyllactose, 3′-SL; 6′-sialyllactose, 6′-SL) were tested against the porcine OSU strain and human RV Wa strain. The RV Wa strain was not inhibited by any oligosaccharides. However, the RV OSU strain infectivity was dose-dependently inhibited by sialic acid (SA)-containing HMO. 3′-SL and 6′-SL concordantly inhibited 125I-radiolabelled RV cellular binding and infectivity/replication. For the in situ study, a midline laparotomy was performed on 21-d-old formula-fed piglets and six 10 cm loops of ileum were isolated in situ. Briefly, 2 mg/ml of LNnT, aHMO mixture (40 % 6′-SL/10 % 3′-SL/50 % SA) or media with or without the RV OSU strain (1 × 107 focus-forming units) were injected into the loops and maintained for 6 h. The loops treated with HMO treatments+RV had lower RV replication, as assessed by non-structural protein-4 (NSP4) mRNA expression, than RV-treated loops alone. In conclusion, SA-containing HMO inhibited RV infectivity in vitro; however, both neutral HMO and SA with aHMO decreased NSP4 replication during acute RV infection in situ.

Probiotics (PRO) modulate immunity in humans, while the effect of prebiotics (PRE) and synbiotics (SYN) on the human immune system are not well studied yet. The objective of this study was to investigate whether daily intake of a SYN modulates immune functions. In a randomised double-blind, placebo-controlled trial, thirty-four colon cancer patients who had undergone ‘curative resection’ and forty polypectomised patients participated. Subjects of the SYN group daily received encapsulated bacteria (1 × 1010 colony-forming units of Lactobacillus rhamnosus GG (LGG) and 1 × 1010 colony-forming units of Bifidobacterium lactis Bb12 (Bb12)) and 10 g of inulin enriched with oligofructose. Controls received encapsulated maltodextrin and 10 g of maltodextrin. Prior to intervention (T1), and 6 (T2) and 12 weeks after the start of the intervention (T3), phagocytic and respiratory burst activity of neutrophils and monocytes, lytic activity of natural killer cells and production of interleukin (IL)-2, IL-10 and IL-12, as well as tumour necrosis factor-α and interferon-γ (IFN-γ) by activated peripheral blood mononuclear cells (PBMC) were measured. In faeces, the concentrations of transforming growth factor-β1 and prostaglandin E2 were measured. IL-2 secretion by activated PBMC from the polyp group increased significantly between T1 or T2 and T3 (P < 0·05). In the cancer group, SYN treatment resulted in an increased capacity of PBMC to produce IFN-γ at T3 (P < 0·05). Other immunity-related parameters were not affected by SYN treatment, neither in the cancer nor in the polyp group. In conclusion, supplementation with this SYN has minor stimulatory effects on the systemic immune system of the two study groups. Further studies in humans should aim to focus on the gut-associated immune system.

The chemosensory components shared by both lingual and intestinal epithelium play a critical role in food consumption and the regulation of intestinal functions. In addition to nutrient signals, other luminal contents, including micro-organisms, are important in signalling across the gastrointestinal mucosa and initiating changes in digestive functions. A potential role of gut microbiota in influencing food intake, energy homeostasis and weight gain has been suggested. However, whether gut microbiota modulates the expression of nutrient-responsive receptors and transporters, leading to altered food consumption, is unknown. Thus, we examined the preference for nutritive (sucrose) and non-nutritive (saccharin) sweet solutions in germ-free (GF, C57BL/6J) mice compared with conventional (CV, C57BL/6J) control mice using a two-bottle preference test. Then, we quantified mRNA and protein expression of the sweet signalling protein type 1 taste receptor 3 (T1R3) and α-gustducin and Na glucose luminal transporter-1 (SGLT-1) of the intestinal epithelium of both CV and GF mice. Additionally, we measured gene expression of T1R2, T1R3 and α-gustducin in the lingual epithelium. We found that, while the preference for sucrose was similar between the groups, GF mice consumed more of the high concentration (8 %) of sucrose solution than CV mice. There was no difference in either the intake of or the preference for saccharin. GF mice expressed significantly more T1R3 and SGLT-1 mRNA and protein in the intestinal epithelium compared with CV mice; however, lingual taste receptor mRNA expression was similar between the groups. We conclude that the absence of intestinal microbiota alters the expression of sweet taste receptors and GLUT in the proximal small intestine, which is associated with increased consumption of nutritive sweet solutions.

PUFA from fish oil appear to have anti-inflammatory and anti-oxidative effects and improve nutritional status in cancer patients. With this as background, the aim of the present study was to investigate the effect of EPA plus DHA on inflammatory condition, and oxidative and nutritional status in patients with lung cancer. In our multicentre, randomised, double-blind trial, thirty-three patients with a diagnosis of advanced inoperable non-small-cell lung cancer and undergoing chemotherapy were divided into two groups, receiving four capsules/d containing 510 mg of EPA and 340 mg of DHA, or 850 mg of placebo, for 66 d. At the start of chemotherapy (T0), after 8 d (T1), 22 d (T2) and 66 d (T3), biochemical (inflammatory and oxidative status parameters) and anthropometric parameters were measured in both groups. A significant increase of body weight in the n-3 group at T3v. T0 was observed. Concerning inflammation, C-reactive protein and IL-6 levels differed significantly between the n-3 and placebo groups at T3, and progressively decreased during chemotherapy in the n-3 group, evidencing n-3 PUFA anti-inflammatory action. Concerning oxidative status, plasma reactive oxygen species levels increased in the placebo group v. the n-3 group at the later treatment times. Hydroxynonenal levels increased in the placebo group during the study, while they stabilised in the n-3 group. Our data confirm that the continual assumption of EPA plus DHA determined an anti-inflammatory and anti-oxidative action which could be considered a preliminary goal in anti-cachectic therapy.

In broiler chickens, a diet where the major cereal types are wheat, rye and/or barley has a lower digestibility compared with a diet in which maize is the major cereal type. In the present study, the effects of two different dietary cereal types, maize v. wheat/rye, on host factors (inflammation and gut integrity) and gut microbiota composition were studied. In addition, the effects of low-dose Zn-bacitracin supplementation were examined. Broilers given a wheat/rye-based diet showed more villus fusion, a thinner tunica muscularis, more T-lymphocyte infiltration, higher amount of immune cell aggregates in the mucosa, more and larger goblet cells and more apoptosis of epithelial cells in the mucosa than those given a maize-based diet. Adding Zn-bacitracin generally reversed these alterations. The microbiota composition was analysed by the use of terminal-restriction fragment length polymorphism, showing changes in the microbiota composition depending on the cereal type used in the diets. The effect of the change of cereal type on the gut microbiota composition was larger than that of Zn-bacitracin supplementation. In conclusion, a wheat/rye-based diet evoked mucosal damage, an alteration in the composition of the microbiota and an inflammatory bowel type of condition.

Dietary protein and amino acid requirement recommendations for normal “healthy” children and adults have varied considerably with 2007 FAO/WHO protein requirement estimates for children lower, but dietary essential AA requirements for adults more than doubled. Requirement estimates as presented do not account for common living conditions, which are prevalent in developing countries such as energy deficit, infection burden and added functional demands for protein and AAs. This study examined the effect of adjusting total dietary protein for quality and digestibility (PDCAAS) and of correcting current protein and AA requirements for the effect of infection and a mild energy deficit to estimate utilizable protein (total protein corrected for biological value and digestibility) and the risk/prevalence of protein inadequacy. The relationship between utilizable protein/prevalence of protein inadequacy and stunting across regions and countries was examined. Data sources (n = 116 countries) included FAO FBS (food supply), UNICEF (stunting prevalence), UNDP (GDP) and UNSTATS (IMR) and USDA nutrient tables. Statistical analyses included Pearson correlations, paired-sample/non-parametric t-tests and linear regression. Statistically significant differences were observed in risk/prevalence estimates of protein inadequacy using total protein and the current protein requirements versus utilizable protein and the adjusted protein requirements for all regions (p < 0·05). Total protein, utilizable protein, GDP per capita and total energy were each highly correlated with the prevalence of stunting. Energy, protein and utilizable protein availability were independently and negatively associated with stunting (p < 0·001), explaining 41 %, 34 % and 40 % of variation respectively. Controlling for energy, total protein was not a statistically significant factor but utilizable protein remained significant explaining~45 % of the variance (p = 0·017). Dietary utilizable protein provides a better index of population impact of risk/prevalence of protein inadequacy than crude protein intake. We conclude that the increased demand for protein due to infections and mild to moderate energy deficits, should be appropriately considered in assessing needs of populations where those conditions still prevail.

Adherence to the Mediterranean diet (MD) has been reported to improve CHD prognosis and to be inversely associated with CHD mortality. The aim of the present study was to investigate the association of adherence to the MD with CHD incidence and mortality in the Greek European Prospective Investigation into Cancer and Nutrition cohort, a population with traditional Mediterranean roots. In a general population sample of 23 929 adult men and women with no CVD or cancer at enrolment, a validated FFQ was interviewer-administered, sociodemographic, physical activity and other characteristics were recorded, and arterial blood pressure and anthropometric characteristics were measured. In a median period of 10 years, 636 incident CHD cases and 240 CHD deaths were recorded. Associations of adherence to the MD, operationalised through a nine-component score (0, poor; 9, excellent), with CHD incidence and mortality were evaluated through Cox regression controlling for potentially confounding variables. A two-point increase in the MD score was associated with lower CHD mortality by 25 % (95 % CI 0·57, 0·98) among women and 19 % (95 % CI 0·67, 0·99) among men. The association of adherence to the MD with CHD incidence was again inverse, but weaker (hazard ratios 0·85 (95 % CI 0·71, 1·02) among women and 0·98 (95 % CI 0·87, 1·10) among men). With respect to score components, only meat among men (positively) and fruits and nuts among women (inversely) were associated with both the incidence of and mortality from CHD. The MD, as an integral entity, is inversely associated with CHD incidence and, particularly, mortality.

1. It has been suggested previously that people in developed countries do not expose themselves to cold severe enough to induce a metabolic response. The energy expenditure, as both heat production and total heat loss, of nine women was therefore measured continuously while each lived for 30 h in a whole-body calorimeter on two occasions, one at 28° and the other at 22°. All subjects followed a predetermined pattern of activity and food intake. The environmental conditions were judged by the subjects to be within those encountered in everyday life. In the standard clothing worn, 28° was considered to be comfortably warm but not too hot, while 22° was judged to be cool but not too cold.

2. Heat production for 24 h was significantly greater at the lower temperature, by (mean ± SE) 7.0 ± 1.1%. The range was between 2 and 12%. Total heat loss was also significantly greater, by 6%, and there was a large change in the partition of heat loss. At the lower temperature sensible heat loss increased by 29% while evaporative heat loss decreased by 39%.

3. Resting metabolism measured in the morning 12–13 h after the last meal was significantly greater at 22° than at 28°, whereas there was no difference when the resting measurement was made for 2.5 h following a meal.

4. In conclusion: (a)environmental temperature may play a more important role than was previously recognized in the energy balance of those living in this country, and (b) there is an indication of at least a partial replacement of cold-induced by diet-induced thermogenesis in man.

1. The extent to which ammonia-N (NH3-N) serves as a starting point for synthesis of microbial nitrogenous compounds was assessed when 15N as (15NH4)2SO4 was continuously infused into the rumen of a sheep for periods of 78–98 h. Steady states were reached in the composition of the rumen contents because the animal was fed equal parts of its ration at hourly intervals. Concentrations of 15N in bacterial-N, protozoal-N and rumen NH3-N were compared.

2. In two trials with a low-N diet consisting largely of wheaten hay the 15N concentration in bacterial N was 76 and 78% of that in the NH3-N. For protozoa the values were more variable —64 and 43%.

In two trials with a higher-N diet (lucerne hay), the corresponding values were lower—bacterial-N 62 and 64%, protozoal-N 41 and 35%.It was concluded that synthesis of microbial protein was more dependent on ammonia as a starting point with the low-N diet than with the higher-N diet.

3. Entry and exit rates for ammonia into and out of the rumen system were determined, and the results, in combination with those obtained for bacterial-N in the first part of the work, allowed calculations to be made of the production of microbial-N/d formed from NH3-N, and this in turn allowed calculation of minimal values for conversion of plant-N to microbial-N in the lumen. Minimal extent of conversion was 68% for the low-N diet and 53–55% for the higher-N diet.

4. Total production of microbial-N in relation to the amount of N given was also calculated by using previously reported values for the relative proportions of protozoal-N and bacterial-N in sheep given diets similar to those used here. These values for extent of conversion were 73% for the low-N diet and 58–59% for the higher-N diet.