Large Neutral Amino Acid Ratio Research Articles

Effect of Varying Levels of Dietary Tryptophan on Aggression and Abnormal Behavior in Growing Pigs.

Body lesions, resulting from tail-biting and ear-biting, can result in decreased health and welfare in pigs. Tryptophan, an indispensable amino acid, is needed to support protein deposition, and the synthesis of the neurotransmitter serotonin, which is important to mood, sleep-wake and eating patterns and might play a role in aggression and abnormal behavior. Two randomized block design studies were conducted to assess the influence of varying dietary tryptophan levels on aggression and abnormal behavior in 8-week-old pigs. Six diets were formulated which met or exceeded all nutrient requirements yet differed according to the dietary tryptophan content. The first study included control (100% standardized ileal digestible tryptophan), supplemented (175% standardized ileal digestible tryptophan), and supplement-plus (250% standardized ileal digestible tryptophan) experimental diets, while the second study included deficient (80% standardized ileal digestible tryptophan), adequate control (105% standardized ileal digestible tryptophan), and extra-tryptophan (130% standardized ileal digestible tryptophan) experimental diets. Concentrations of plasma tryptophan and large neutral amino acids (tyrosine, isoleucine, leucine, valine, and phenylalanine) were analyzed using ultra-performance liquid chromatography and the tryptophan to large neutral amino acid ratio was calculated. Analysis for time active, lying, and engaging in aggressive interactions was carried out using 10-min scan samples to determine behavioral time budgets of the pigs on different experimental diets. Pigs fed diets with supplemented tryptophan had higher concentrations of both plasma tryptophan and tryptophan to large neutral amino acid ratio compared to the pigs fed the control diet (P < 0.05) in the first study, while no significant differences were detected for plasma tryptophan or the tryptophan to large neutral amino acid ratio in the second study. Diet did not have an effect (P > 0.05) on weight, feed intake or behavior throughout the studies. The results suggest that an increase in dietary tryptophan relative to large neutral amino acids, fed for 29 days, impacts circulating plasma tryptophan and therefore, serotonin concentrations in the pig. Despite an increase in circulating plasma tryptophan in response to an increase in dietary tryptophan in the first study, we failed to see an impact of the dietary treatment on body, tail and ear-biting behavior under the conditions studied.

Association Between Dietary Protein Intake and Sleep Quality in Middle-Aged and Older Adults in Singapore.

Poor sleep has been associated with the increased risk of developing detrimental health conditions. Diet and certain nutrients, such as dietary protein (PRO) may improve sleep. This cross-sectional study aimed to investigate the relationship between PRO intake, their amino acid components, and sources with sleep quality in middle-aged and older adults residing in Singapore. A dataset of 104 healthy subjects between the age of 50 and 75 years old were used. Collected data included 3-day food record and sleep quality [sleep duration, global sleep score (GSS), sleep latency (SL), and sleep efficiency (SE)]. The collected 3-day food records were extracted for PRO, tryptophan (Trp), and large neutral amino acid (LNAA) intake. PRO intake was further categorized into plant and animal PRO. A multivariate multiple linear regression (MLR) was performed to assess the association between PRO intake and sleep quality. Dietary Trp:LNAA ratio was positively associated with sleep duration (βtotal: 108.234 h; p: 0.005) after multiple covariates adjustment. Similarly, plant Trp (βplant: 2.653 h/g; p: 0.020) and plant Trp:LNAA (βplant: 54.006 h; p: 0.008) was positively associated with sleep duration. No significant associations were observed for both SL and SE. Sleep duration in middle-aged and older Singaporean adults was positively associated with dietary Trp and Trp:LNAA, especially when obtained from plant sources.

Eating to dare - Nutrition impacts human risky decision and related brain function

Macronutrient composition modulates plasma amino acids that are precursors of neurotransmitters and can impact brain function and decisions. Neurotransmitter serotonin has been shown to regulate not only food intake, but also economic decisions. We investigated whether an acute nutrition-manipulation inducing plasma tryptophan fluctuation affects brain function, thereby affecting risky decisions. Breakfasts differing in carbohydrate/protein ratios were offered to test changes in risky decision-making while metabolic and neural dynamics were tracked. We identified that a high-carbohydrate/protein breakfast increased plasma tryptophan/LNAA (large neutral amino acids) ratio which mapped to individual risk propensity changes. The nutrition-manipulation and tryptophan/LNAA fluctuation effects on risk propensity changes were further modulated by individual differences in body fat mass. Using fMRI, we further identified activation in the parietal lobule during risk-processing, of which activities 1) were sensitive to the tryptophan/LNAA fluctuation, 2) were modulated by individual's body fat mass, and 3) predicted the risk propensity changes in decision-making. Our results provide evidence for a personalized nutrition-driven modulation on human risky decision and its metabolic and neural mechanisms.

Investigating the effects of incremental conditioning and supplemental dietary tryptophan on the voluntary activity and behaviour of mid-distance training sled dogs.

Serotonin is a neurotransmitter synthesized by the amino acid tryptophan, that has the potential to impact the behaviour and activity of dogs. The objective of this study was to assess the effects of supplemental tryptophan and a 12-week incremental training regimen on the voluntary activity and behaviour of client-owned Siberian Huskies. Sixteen dogs were blocked for age, BW and sex and then randomly allocated to either the control or treatment group. Both groups were fed the same dry extruded diet; however, the treatment group were supplemented with tryptophan to achieve a tryptophan: large neutral amino acid ratio of 0.075:1. Once a week, a 5-minute video recording was taken immediately pre- and post- exercise to evaluate dogs’ behaviours. Activity monitors were used to record voluntary activity on both training and rest days. Linear regression analysis was used to assess the relationship between training week and time spent performing each behaviour. Additionally, a repeated measure mixed model was used to test differences between diet groups and training week for both behavioural and activity count data. The time spent performing agonistic behaviours prior to exercise was negatively associated with week for treatment dogs (β = -0.32, 95% CI [-0.55, -0.10], P < 0.05) and no change was observed for control dogs (β = -0.13, 95% CI [-0.41, 0.15], P > 0.10). Treatment did not have any effect on activity levels (P > 0.10). For all dogs, locomotive behaviours decreased prior to exercise as weeks progressed (P < 0.05), while run day voluntary activity depended on the distance run that day (P < 0.05). These data suggest that sled dogs experience an exercise-induced reduction in voluntary locomotion in response to both single bouts and repetitive bouts of exercise. Additionally, tryptophan supplementation may decrease agonistic behaviours, without having any effect on voluntary activity.

Exercise but Not Supplemental Dietary Tryptophan Influences Heart Rate and Respiratory Rate in Sled Dogs.

Tryptophan (Trp), an indispensable amino acid for dogs, is the precursor of serotonin, a neurotransmitter with a variety of effects throughout the body, including the ability to modulate cardiac and pulmonary activity. This study aimed to investigate the effects of a 12-week incremental exercise regimen and supplemental dietary Trp on heart rate (HR) and respiratory rate (RR) in client-owned sled dogs. Sixteen Siberian huskies were randomly allocated to either treatment or control diet groups. Both groups were fed a control diet (Trp to large neutral amino acid ratio of 0.047:1); however, treatment dogs received a Trp supplement to achieve a Trp to large neutral amino acid ratio of 0.075:1. Every three weeks, external telemetry equipment was used to non-invasively measure and record HR and RR at a resting, working, and post-exercise state in a controlled exercise challenge. A mixed model was used to test differences between diet, activity parameter, and week. Dietary Trp supplementation had no effect on HR or RR. Independent of diet, resting, working, post-exercise HR, and time to recover post-exercise HR decreased from week −1 to week 11 (p < 0.05). Resting HR had the greatest reduction from week −1 to week 11 (21%, p < 0.05). Working RR did not change with exercise (p > 0.10), but rRR and postRR decreased from week −1 to week 11 (p < 0.05). These data suggest that the exercise regimen the dogs were subjected to may have positively impacted the dogs’ capacity to sustain aerobic exercise, whereas Trp supplementation had no effect on HR or RR.

Randomized Double-Blind Controlled Trial on the Effect of Proteins with Different Tryptophan/Large Neutral Amino Acid Ratios on Sleep in Adolescents: The PROTMORPHEUS Study.

Disturbed sleep is common in adolescents. Ingested nutrients help regulate the internal clock and influence sleep quality. The purpose of this clinical trial is to assess the effect of protein tryptophan (Trp)/large neutral amino acids (LNAAs) ratio on sleep and circadian rhythm. Ingested Trp is involved in the regulation of the sleep/wake cycle and improvement of sleep quality. Since Trp transport through the blood–brain barrier is competing with LNAAs, protein with higher Trp/LNAAs were expected to increase sleep efficiency. This randomized double-blind controlled trial will enroll two samples of male adolescents predisposed to sleep disturbances: elite rugby players (n = 24) and youths with obesity (n = 24). They will take part randomly in three sessions each held over a week. They will undergo a washout period, when dietary intake will be calibrated (three days), followed by an intervention period (three days), when their diet will be supplemented with three proteins with different Trp/LNAAs ratios. Physical, cognitive, dietary intake, appetite, and sleepiness evaluations will be made on the last day of each session. The primary outcome is sleep efficiency measured through in-home electroencephalogram recordings. Secondary outcomes include sleep staging, circadian phase, and sleep-, food intake-, metabolism-, and inflammation-related biochemical markers. A fuller understanding of the effect of protein Trp/LNAAs ratio on sleep could help in developing nutritional strategies addressing sleep disturbances.

Effects of incremental exercise and dietary tryptophan supplementation on the amino acid metabolism, serotonin status, stool quality, fecal metabolites, and body composition of mid-distance training sled dogs.

Exercise improves the health of dogs; however, the extreme exertion experienced by sled dogs may lead to variable metabolic and fecal characteristics. Nutritional interventions, such as dietary tryptophan (Trp), may reduce the prevalence of these exercise-induced disturbances. Sporting diets tend to have high crude protein concentrations in contrast to adult maintenance diets and this results in less Trp relative to other amino acids (AA). Therefore, sporting dogs represent an ideal cohort to assess the effects of supplemental Trp. The objective was to evaluate the effects of supplemental dietary Trp and an incremental training regimen on AA and serotonin status, fecal scores and metabolites, and body composition in client-owned Siberian huskies. Sixteen dogs (nine females and seven males) were used, with a mean age of 4.8 ± 2.5 yr and body weight (BW) of 24.3 ± 4.3 kg. Dogs were blocked for sex, age, and BW and randomly allocated into two groups with eight fed a dry extruded control diet (Ctl) and eight fed Ctl supplemented with Trp to reach a Trp:large-neutral AA (LNAA) ratio of 0.075:1 (treatment, Trt). The exercise regimen was designed to increase in distance each week, but weather played a role in setting the daily distance. Each week BW was recorded and food allotments were adjusted to maintain initial BW. Pre and post-exercise blood samples were taken every 3 wk, dogs then received a meal followed by 1, 2, and 4 h post meal blood collections (serum AA, serotonin). Stool collection and scoring occurred each week and body composition was measured on weeks -1 and 11. Serotonin, AA, fecal metabolite, and body composition data were analyzed using PROC MIXED of SAS with dog as a random effect and week and Trt as fixed effects. Stool score data were analyzed using PROC FREQ to compare stool score and Trt, and PROC CORR was used to analyze associations between fecal score, temperature, humidity, and run distance. Dogs on Trt had greater fasted Trp compared with baseline, greater post-meal Trp and serotonin compared with baseline, greater post-meal Trp compared with fasted, and greater post-meal Trp and serotonin compared with Ctl (P < 0.05). Fecal data indicated that Trp improved stool scores (P < 0.05) yet had no effect on fecal metabolites. An overall increase in lean and decrease in fat mass was found (P < 0.05), but Trt had no effect on body composition. Optimization of the dietary Trp:LNAA ratio may help to improve GI health without compromising performance in actively training sled dogs.

Effects of intragastric tryptophan on acute changes in the plasma tryptophan/large neutral amino acids ratio and relationship with subsequent energy intake in lean and obese men.

Circulating tryptophan/large neutral amino acids (tryptophan/LNAA) ratio, an indicator of brain serotonin levels, may be important in appetite regulation, together with gastrointestinal (gastric emptying, plasma cholecystokinin) mechanisms. We have compared effects of intragastric tryptophan ('Trp') on the plasma tryptophan/LNAA ratio in lean and obese men, and the associations of the tryptophan/LNAA ratio, gastric emptying and CCK concentrations with energy intake. Lean and obese male participants (n = 16 each) received 3 g Trp or volume-matched control intragastrically, 15 min before a mixed-nutrient drink (300 mL, 400 kcal) (t = 0 min) in randomised, double-blind fashion. Plasma amino acid (for calculation of the plasma tryptophan/LNAA ratio) and CCK concentrations were measured from t = -20-60 min. Gastric emptying was assessed from t = 0-60 min, and ad-libitum energy intake from a standardised buffet-style meal from t = 60-90 min. The increase in the plasma tryptophan/LNAA ratio was less in obese, than lean, participants (P < 0.05), and greater in lean participants who reduced their energy intake (by >0 kcal) after Trp compared with those who did not (by ≤0 kcal) (P < 0.05). Moreover, in participants who reduced their energy intake, the ratio was lower in obese, than in lean (P < 0.05). There was a trend for an inverse correlation between energy intake with the plasma tryptophan/LNAA ratio in lean (r = -0.4, P = 0.08), but not in obese, participants. There was no significant difference in gastric emptying or CCK between participants who reduced their energy intake and those who did not. In conclusion, the plasma tryptophan/LNAA ratio appears to be a determinant of the suppression of energy intake in response to tryptophan in normal-weight people, but not in those with obesity. The role of the plasma tryptophan/LNAA ratio to regulate energy intake, and potential changes in obesity, warrant evaluation in prospective studies.

Effect of forage type on cecal and fecal microbiome and fermentation parameters in the horse

Tryptophan (Trp) is an amino acid used as precursor for several molecules in the body, including serotonin. Serotonin is involved in a broad range of physiological and behavioral functions, including stress response. Because serotonin cannot cross the blood-brain barrier, supply of Trp to the brain could be used to elevate serotonin synthesis and release. Evidence in monogastric species indicates that Trp supplementation may increase circulating serotonin and reduces stress response, but there is conflicting evidence in the literature about degradation of Trp in the rumen. Some authors indicated that Trp is degraded slowly in the rumen and that it may remain available to be absorbed, whereas others observed that it is quickly degraded by rumen bacteria. Thus, the goal of the experiment was to test if supplementation of free L-Trp in steers may increase plasma Trp and serotonin, thus changing behavior and reducing circulating concentrations of hormones that are related to stress response. Four hybrid steers were used in a Latin square design with supplementation of L-Trp at 0, 50, 100, and 150 mg/kg BW/d for one week and three weeks as washout period. Blood samples were analyzed for amino acids, adrenocorticotropic hormone, adrenaline, cortisol, serotonin, and glucose. Videos were used to evaluate posture and activities every 10 minutes. Results indicated no effect of diet or diet by day interaction for most of the evaluated variables and activities. The only difference detected was a diet effect on time spent inactive. Steers receiving 150 mg Trp/kg BW spent more time inactive than steers in the control (69.51 ± 2.01 vs. 60.9 ± 1.92; P = 0.0305). We concluded that it was not possible to raise plasma Trp in steers using up to 150 mg/kg BW/d free Trp and that higher doses might be needed to increase plasma Trp and Trp:large neutral amino acid ratio, and to induce meaningful behavioral changes in steers.

Changes in behavior and plasma metabolites after tryptophan supplementation in steers

Tryptophan (Trp) is an amino acid used as precursor for several molecules in the body, including serotonin. Serotonin is involved in a broad range of physiological and behavioral functions, including stress response. Because serotonin cannot cross the blood-brain barrier, supply of Trp to the brain could be used to elevate serotonin synthesis and release. Evidence in monogastric species indicates that Trp supplementation may increase circulating serotonin and reduces stress response, but there is conflicting evidence in the literature about degradation of Trp in the rumen. Some authors indicated that Trp is degraded slowly in the rumen and that it may remain available to be absorbed, whereas others observed that it is quickly degraded by rumen bacteria. Thus, the goal of the experiment was to test if supplementation of free L-Trp in steers may increase plasma Trp and serotonin, thus changing behavior and reducing circulating concentrations of hormones that are related to stress response. Four hybrid steers were used in a Latin square design with supplementation of L-Trp at 0, 50, 100, and 150 mg/kg BW/d for one week and three weeks as washout period. Blood samples were analyzed for amino acids, adrenocorticotropic hormone, adrenaline, cortisol, serotonin, and glucose. Videos were used to evaluate posture and activities every 10 minutes. Results indicated no effect of diet or diet by day interaction for most of the evaluated variables and activities. The only difference detected was a diet effect on time spent inactive. Steers receiving 150 mg Trp/kg BW spent more time inactive than steers in the control (69.51 ± 2.01 vs. 60.9 ± 1.92; P = 0.0305). We concluded that it was not possible to raise plasma Trp in steers using up to 150 mg/kg BW/d free Trp and that higher doses might be needed to increase plasma Trp and Trp:large neutral amino acid ratio, and to induce meaningful behavioral changes in steers.

Effects of tryptophan depletion on selective serotonin reuptake inhibitor-remitted patients with obsessive compulsive disorder.

Serotonergic antidepressants are first-line medication therapies for obsessive-compulsive disorder, however it is not known if synaptic serotonin availability is important for selective serotonin reuptake inhibitor efficacy. The present study tested the hypothesis that temporary reduction in central serotonin transmission, through acute tryptophan depletion, would result in an increase in anxiety in selective serotonin reuptake inhibitor-remitted obsessive-compulsive disorder patients. Eight patients (four males) with obsessive-compulsive disorder who showed sustained clinical improvement with selective serotonin reuptake inhibitor treatment underwent acute tryptophan depletion in a randomized, double-blind, placebo-controlled, within-subjects design, over two days one week apart. Five hours after consumption of the depleting/sham drink the participants performed a personalized obsessive-compulsive disorder symptom exposure task. Psychological responses were measured using the Spielberger State Anxiety Inventory, Yale-Brown Obsessive Compulsive Scale and Visual Analogue Scales. Free plasma tryptophan to large neutral amino acid ratio decreased by 93% on the depletion day and decreased by 1% on the sham day, as anticipated. Psychological rating scores as measured by Visual Analogue Scale showed a significant decrease in perceived control and increase in interfering thoughts at the time of provocation on the depletion day but not on the sham day. A measure of convergent validity, namely Visual Analogue Scale Similar to past, was significantly higher at the time of provocation on both the depletion and sham days. Both the depletion and time of provocation scores for Visual Analogue Scale Anxiety, Spielberger State Anxiety Inventory, Yale-Brown Obsessive Compulsive Scale and blood pressure were not significant. Acute tryptophan depletion caused a significant decrease in perceived control and increase in interfering thoughts at the time of provocation. Acute tryptophan depletion had no effect on the Spielberger State Anxiety Inventory or Visual Analogue Scale Anxiety measures, which suggests that the mechanism of action of selective serotonin reuptake inhibitors may be different to that seen in panic, social anxiety and post-traumatic stress disorder. Successful selective serotonin reuptake inhibitor treatment of obsessive-compulsive disorder may involve the ability of serotonin to switch habitual responding to goal-directed behaviour.

Breakfast high in whey protein or carbohydrates improves coping with workload in healthy subjects

Dietary components may affect brain function and influence behaviour by inducing the synthesis of neurotransmitters. The aim of the present study was to examine the influence of consumption of a whey protein-containing breakfast drink v. a carbohydrate drink v. control on subjective and physiological responses to mental workload in simulated work. In a randomised cross-over design, ten healthy subjects (seven women, median age 26 years, median BMI 23 kg/m(2)) participated in a single-blinded, placebo-controlled study. The subjects performed demanding work-like tasks after having a breakfast drink high in protein (HP) or high in carbohydrate (HC) or a control drink on separate sessions. Subjective states were assessed using the NASA Task Load Index (NASA-TLX), the Karolinska sleepiness scale (KSS) and the modified Profile of Mood States. Heart rate was recorded during task performance. The ratio of plasma tryptophan (Trp) to the sum of the other large neutral amino acids (LNAA) and salivary cortisol were also analysed. The plasma Trp:LNAA ratio was 30 % higher after the test drinks HP (median 0·13 (μmol/l)/(μmol/l)) and HC (median 0·13 (μmol/l)/(μmol/l)) than after the control drink (median 0·10 (μmol/l)/(μmol/l)). The increase in heart rate was smaller after the HP (median 2·7 beats/min) and HC (median 1·9 beats/min) drinks when compared with the control drink (median 7·2 beats/min) during task performance. Subjective sleepiness was reduced more after the HC drink (median KSS - 1·5) than after the control drink (median KSS - 0·5). There were no significant differences between the breakfast types in the NASA-TLX index, cortisol levels or task performance. We conclude that a breakfast drink high in whey protein or carbohydrates may improve coping with mental tasks in healthy subjects.

Determining the effect of the ratio of tryptophan to large neutral amino acids on the growth performance of finishing pigs (2011)

A total of 96 pigs (PIC TR4 × 1050; initially 77.4 lb) were used in 2 14-d studies to determine the effect of standardized ileal digestible (SID) tryptophan to large neutral amino acids (LNAA) ratio on growth performance of finishing pigs. Pens of pigs were balanced by initial weight and randomly allotted to 1 of 4 dietary treatments in a completely randomized design with 4 pigs per pen and 6 replications per treatment. The treatment diets were fed in 2 phases: early finishing phase (77 to 106 lb BW) and late finishing phase (183 to 217 lb BW), with a common diet fed in between. Dietary treatments included: (1) a corn-soybean meal-based diet without DDGS, (2) a cornsoybean meal-based diet with 45% dried distillers grains with solubles (DDGS), (3) a corn-soybean meal-based diet without DDGS but supplemented with similar amounts of LNAA as the diet containing 45% DDGS, and (4) the LNAA-supplemented diet with added crystalline tryptophan to increase the SID tryptophan:LNAA ratio. The diets were formulated in a similar manner for the late finishing phase with the exception that DDGS were lowered to 30% of the diet. In the early finishing period (77 to 106 lb), pigs fed 45% DDGS diet had poorer F/G (P = 0.01) compared with pigs fed the other diets; however, no differences were found in other response criteria. During the late finishing period (183 to 217 lb), pig growth performance was not affected by dietary treatment. These results suggest that the high level of LNAA relative to tryptophan in diets containing 30% DDGS or greater may not be responsible for the apparent increase in the tryptophan requirement of finishing pigs seen in previous studies.

Effects of an acute α-lactalbumin manipulation on mood and food hedonics in high- and low-trait anxiety individuals

Serotonergic hypofunction is associated with a depressive mood state, an increased drive to eat and preference for sweet (SW) foods. High-trait anxiety individuals are characterised by a functional shortage of serotonin during stress, which in turn increases their susceptibility to experience a negative mood and an increased drive for SW foods. The present study examined whether an acute dietary manipulation, intended to increase circulating serotonin levels, alleviated the detrimental effects of a stress-inducing task on subjective appetite and mood sensations, and preference for SW foods in high-trait anxiety individuals. Thirteen high- (eleven females and two males; anxiety scores 45.5 (sd 5.9); BMI 22.9 (sd 3.0)kg/m(2)) and twelve low- (ten females and two males; anxiety scores 30.4 (sd 4.8); BMI 23.4 (sd 2.5) kg/m(2)) trait anxiety individuals participated in a placebo-controlled, two-way crossover design. Participants were provided with 40 g alpha-lactalbumin (LAC; l-tryptophan (Trp):large neutral amino acids (LNAA) ratio of 7.6) and 40 g casein (placebo) (Trp:LNAA ratio of 4.0) in the form of a snack and lunch on two test days. On both the test days, participants completed a stress-inducing task 2 h after the lunch. Mood and appetite were assessed using visual analogue scales. Changes in food hedonics for different taste and nutrient combinations were assessed using a computer task. The results demonstrated that the LAC manipulation did not exert any immediate effects on mood or appetite. However, LAC did have an effect on food hedonics in individuals with high-trait anxiety after acute stress. These individuals expressed a lower liking (P = 0.012) and SW food preference (P = 0.014) after the stressful task when supplemented with LAC.

Brain serotonergic and dopaminergic modulators, perceptual responses and endurance exercise performance following caffeine co-ingested with a high fat meal in trained humans

BackgroundThe present study examined putative modulators and indices of brain serotonergic and dopaminergic function, perceptual responses, and endurance exercise performance following caffeine co-ingested with a high fat meal.MethodsTrained humans (n = 10) performed three constant-load cycling tests at 73% of maximal oxygen uptake (VO2max) until exhaustion at 10°C remove space throughout. Prior to the first test, subjects consumed a 90% carbohydrate meal (Control trial) and for the remaining two tests, a 90% fat meal with (FC trial) and without (F trial) caffeine.ResultsTime to exhaustion was not different between the F and FC trials (P > 0.05); [Control trial: 116(88-145) min; F trial: 122(96-144) min; FC trial: 127(107-176) min]. However, leg muscular discomfort during exercise was significantly lower on the FC relative to F trial (P < 0.01). There were no significant differences between F and FC trials in key modulators and indices of brain serotonergic (5-HT) and dopaminergic (DA) function [(i.e. plasma free and total tryptophan (Trp), tyrosine (Tyr), large neutral amino acids (LNAA), Trp:LNAA ratio, free-Trp:Tyr ratio, total Trp:Tyr ratio, and plasma prolactin] (P > 0.05) with the exception of plasma free-Trp:LNAA ratio which was higher at 90 min and at exhaustion during the FC trial (P < 0.05).ConclusionsNeither brain 5-HT nor DA systems would appear to be implicated in the fatigue process when exercise is performed without significant thermoregulatory stress, thus indicating fatigue development during exercise in relatively cold temperatures to occur predominantly due to glycogen depletion.

Acute tryptophan depletion potentiates 3,4‐methylenedioxymethamphetamine‐induced cerebrovascular hyperperfusion in adult male wistar rats

The serotonergic (5-hydroxytryptamine; 5-HT) dysfunction found in depression may affect not only brain function (mood) but also cerebrovascular control. Similar, but possibly occult, disturbances may also be induced by 3,4-methylenedioxymethamphetamine-induced neurotoxicity (MDMA, or "ecstasy"). Acute tryptophan depletion (ATD) is widely used to identify vulnerability to depression, and we hypothesized that repeated MDMA administration would increase the sensitivity of rats to this acute serotonergic challenge. In this study, male Wistar rats were injected with MDMA (20 mg kg(-1), twice daily for 4 days) and challenged 3 weeks later with ATD, induced by intragastric administration of a nutritional mixture with tryptophan (TRP) removed. Cerebral metabolism (CMRG) and blood flow (CBF) were measured in parallel groups of animals following ATD by using quantitative [(14)C]2-deoxyglucose and [(14)C]iodoantipyrine autoradiographic techniques, respectively. A significant reduction in paroxetine binding to 5-HT transporter sites in MDMA-treated rats indicated 5HT terminal depletion, whereas the plasma TRP/sum large neutral amino acids ratio was reduced by 40% following ATD. Under all experimental conditions, the normal close correlation between CBF and metabolic demand was maintained. However, a global analysis of all brain regions revealed a significant decrease in the overall ratio of CBF to CMRG after ATD in control animals, whereas a higher ratio was observed after ATD in the MDMA-treated group. This increase in blood flow relative to cerebral metabolism suggests an ATD-induced loss of cerebrovascular tone in MDMA-treated animals that could have pathophysiological consequences and might conceivably contribute to the behavioral dysfunction of depression.

Effects of acute tryptophan depletion in serotonin reuptake inhibitor-remitted patients with generalized anxiety disorder

Serotonergic antidepressants [selective serotonin reuptake inhibitor (SSRI)] are first-line treatments for generalised anxiety disorder (GAD); however, it is not known if synaptic serotonin (5-HT) availability is important for SSRI efficacy. The present study tested the hypothesis that temporary reduction in central 5-HT transmission, through acute tryptophan depletion (ATD), would reverse the therapeutic effect of the SSRIs in GAD patients. Twelve patients (six males) with GAD, who showed sustained clinical improvement with SSRI treatment, underwent ATD in a double-blind, placebo-controlled, within-subjects design over 2 days, 1 week apart. At the peak time of depletion, the participants inhaled 7.5% CO2 and air in random order for at least 12 min each. Psychological responses were measured using the Spielberger State Anxiety Inventory (STAI-S) and GAD-symptom visual analogue scales (VASs; e.g., worry and tense) and Profile of Mood States. Free plasma tryptophan to large neutral amino acid (LNAA) ratio decreased by 92% on the depletion day and decreased by 2% on the control day. Irrespective of depletion condition, 7.5% CO(2) inhalation significantly increased STAI-S and GAD-related VAS scores (all p < 0.05) compared with air inhalation. ATD had no effect on any of these measures despite the substantial reduction in free tryptophan/LNAA ratio. Although SSRIs treat GAD effectively, the present results suggest that the mechanism of action is different to that seen in panic, social anxiety, and post-traumatic stress disorders. Successful SSRI treatment of GAD may involve long-term receptor changes or alterations in other neurotransmitter systems downstream of serotonin.

Plasma tryptophan and tyrosine levels are independent risk factors for delirium in critically ill patients

The pathophysiology of delirium remains elusive though neurotransmitters and their precursor large neutral amino acids (LNAAs) may play a role. This pilot study investigated whether alterations of tryptophan (Trp), phenylalanine (Phe), and tyrosine (Tyr) plasma levels were associated with a higher risk of transitioning to delirium in critically ill patients. Plasma LNAA concentrations were determined on days 1 and 3 in mechanically ventilated (MV) patients from the MENDS randomized controlled trial (dexmedetomidine vs. lorazepam sedation). Three independent variables were calculated by dividing plasma concentrations of Trp, Phe, and Tyr by the sum of all other LNAA concentrations. Delirium was assessed daily using the confusion assessment method for the intensive care unit (CAM-ICU). Markov regression models were used to analyze independent associations between plasma LNAA ratios and transition to delirium after adjusting for covariates. The 97 patients included in the analysis had a high severity of illness (median APACHE II, 28; IQR, 24-32). After adjusting for confounders, only high or very low tryptophan/LNAA ratios (p = 0.0003), and tyrosine/LNAA ratios (p = 0.02) were associated with increased risk of transitioning to delirium, while phenylalanine levels were not (p = 0.27). Older age, higher APACHE II scores and increasing fentanyl exposure were also associated with higher probabilities of transitioning to delirium. In this pilot study, plasma tryptophan/LNAA and tyrosine/LNAA ratios were associated with transition to delirium in MV patients, suggesting that alterations of amino acids may be important in the pathogenesis of ICU delirium. Future studies evaluating the role of amino acid precursors of neurotransmitters are warranted in critically ill patients.

Acute effects of breakfasts containing α-lactalbumin, or gelatin with or without added tryptophan, on hunger, ‘satiety’ hormones and amino acid profiles

Proteins are the most satiating macronutrients. Tryptophan (TRP) may contribute to the satiating effect, as it serves as a precursor for the anorexigenic neurotransmitter serotonin. To address the role of TRP in the satiating properties of dietary protein, we compared three different breakfasts, containing either alpha-lactalbumin (high in TRP), gelatin (low in TRP) or gelatin with added TRP (gelatin+TRP, high in TRP), on appetite. Twenty-four subjects (22-29 kg/m2; aged 19-37 years) received a subject-specific breakfast at t = 0 with 10, 55 and 35 % energy from protein, carbohydrate and fat respectively in a randomised, single-blind design. Hunger, glucagon-like peptide (GLP)-1, ghrelin, amino acid concentrations and energy intake during a subsequent lunch were determined. Suppression of hunger was stronger 240 min after the breakfast with alpha-lactalbumin compared with gelatin and gelatin+TRP. Total plasma amino acid concentrations were lower with alpha-lactalbumin compared with gelatin with or without TRP (from t = 180-240 min). TRP concentrations were higher after alpha-lactalbumin than after gelatin with or without TRP from t = 0-100 min, whereas from t = 100-240 min, TRP concentrations were lower after gelatin than after alpha-lactalbumin and gelatin+TRP. The plasma ratio of TRP to other large neutral amino acids (LNAA) was, only at t = 100 min, lower after gelatin+TRP than after the other breakfasts. Plasma amino acid responses, TRP concentrations and TRP:LNAA ratios were not correlated with hunger. GLP-1 and ghrelin concentrations were similar for all diets. Energy intake during a subsequent lunch was similar for all diets. Summarised, an alpha-lactalbumin breakfast suppresses hunger more than a gelatin or gelatin+TRP breakfast. This cannot be explained by (possible) differences found in TRP concentrations and TRP:LNAA ratios in the breakfasts and in plasma, as well as in circulating total amino acids, GLP-1 and ghrelin.

Changes in Plasma Amino Acid Levels Do Not Predict Satiety and Weight Loss on Diets with Modified Macronutrient Composition

Objective: Serotonin mediates satiety in the central nervous system. Brain serotonin content depends on the plasma ratio of tryptophan (Trp) to large neutral amino acids (LNAA) and may be affected by diet composition. We examined whether high-carbohydrate or high-protein diets induce satiety and weight loss by altering plasma concentrations of these amino acids. Methods: In study 1 (n = 16, BMI = 27.0 ± 2.3), we compared plasma Trp and LNAA concentrations averaged over 24 h after 2 weeks of consuming isocaloric diets containing either 45 or 65% of total energy as carbohydrate. In study 2 (n = 19, BMI = 26.2 ± 2.1), we made the same measurements following diets containing either 15 or 30% of total energy as protein. To assess satiety in both studies, we recorded caloric intake and weight changes during a subsequent 12-week period of ad libitum consumption of the experimental diets. Results: Ad libitum caloric intake fell by 222 ± 81 kcal/day with a 3.7 ± 0.6 kg weight loss at 12 weeks in study 1. Ad libitum caloric intake fell by 441 ± 63 kcal/ day with a 4.9 ± 0.5 kg weight loss at 12 weeks in study 2. The 24-hour averaged plasma concentration of Trp and the Trp:LNAA ratio were unaffected by the isocaloric increase in carbohydrate or protein consumption that preceded the ad libitum administration of the 2 diets. Conclusion: An increase in either carbohydrate or protein intake increases satiety and leads to significant weight loss, however, these effects are not mediated by an increase in plasma concentration of Trp or the Trp:LNAA ratio.