Volatile Short-Chain Aliphatic Aldehydes Act as Taste Modulators through the Orally Expressed Calcium-Sensing Receptor CaSR.

Aldehydes are natural volatile aroma compounds generated by the Maillard reaction of sugars and amino acids in food and affect the flavor of food. They have been reported to exert taste-modifying effects, such as increases in taste intensity at concentrations below the odor detection threshold. The present study examined the taste-enhancing effects of short-chain aliphatic aldehydes, such as isovaleraldehyde (IVAH) and 2-methylbutyraldehyde, thus attempting to identify the taste receptors involved. The results obtained revealed that IVAH enhanced the taste intensity of taste solutions even under the condition of olfactory deprivation by a noseclip. Furthermore, IVAH activated the calcium-sensing receptor CaSR in vitro. Receptor assays on aldehyde analogues showed that C3-C6 aliphatic aldehydes and methional, a C4 sulfur aldehyde, activated CaSR. These aldehydes functioned as a positive allosteric modulator for CaSR. The relationship between the activation of CaSR and taste-modifying effects was investigated by a sensory evaluation. Taste-modifying effects were found to be dependent on the activation state of CaSR. Collectively, these results suggest that short-chain aliphatic aldehydes function as taste modulators that modify sensations by activating orally expressed CaSR. We propose that volatile aroma aldehydes may also partially contribute to the taste-modifying effect via the same molecular mechanism as kokumi substances.

Effects of TUG-891, a potent GPR120 agonist, on the physical and oral lipid- coating properties, and secretion of saliva.

GPR120 agonists were recently shown to enhance the fatty orosensation in humans when added to vegetable oil or a low-fat food system, but did not evoke it by themselves. Furthermore, an emulsion prepared from vegetable oil had a stronger fatty orosensation than that prepared from mineral oil even though the physical properties of both emulsions were similar. To clarify the mechanisms underlying the enhancement of the fatty orosensation by GPR120 agonists, the present study investigated the effects of TUG-891, a potent GPR120 agonist, on physical and oral lipid-coating properties and the secretion of saliva. The addition of TUG-891 to a vegetable oil emulsion did not significantly change its physical properties, such as viscosity, particle distribution, interfacial tension, contact angle, frictional load, and ζ-electric potential, or the amount of the lipid coating remaining in the oral cavity. These results indicate that TUG-891 enhanced the fatty orosensation without changing the physical or oral lipid-coating properties of the emulsion. The addition of TUG-891 to a vegetable oil emulsion and whipped cream significantly increased the amount of saliva secreted. Therefore, TUG-891, a potent GPR120 agonist, may enhance the fatty orosensation by increasing the amount of saliva secreted.

Increases in the pungency of allyl isothiocyanate and piperine by CaSR agonists, glutathione and γ-glutamyl-valyl-glycine.

γ-Glutamyl peptides, including glutathione (γ-Glu-Cys-Gly, GSH) and γ-glutamyl-valyl-glycine (γ-Glu-Val-Gly), have been shown to increase the intensity of basic tastes, such as salty, sweet, and umami, and flavor, including mouthfulness, but had no taste themselves at the concentrations tested. Although the mechanisms of action of γ-glutamyl peptides currently remain unclear, the involvement of the calcium sensing receptor (CaSR) has been suggested. Since GSH and γ-Glu-Val-Gly increase the pungency of some spices, the present study investigated their effects on the pungency of allyl isothiocyanate (AITC) using a sensory evaluation. GSH and γ-Glu-Val-Gly both significantly increased the pungency of AITC, while anserine, a peptide without CaSR activity, did not. GSH-induced increases in pungency were suppressed by NPS-2143, a CaSR inhibitor. Further, γ-Glu-Val-Gly significantly increased the pungency of piperine. The present results suggest that GSH and γ-Glu-Val-Gly increased the pungency by activating CaSR.

GPR120 agonists enhance the fatty orosensation when added to fat-containing system, but do not evoke it by themselves in humans.

Dietary fat, an important macronutrient, has been considered to be perceived by texture and olfaction. Recently, fatty acid transporter, CD36, and fatty acid receptor, GPR120 are considered to be involved in human gustatory fatty acids perception in humans. However, limited information is currently available to show that agonists of CD36 and GPR120 evoke fatty oral sensations regarding to dietary fat in humans. Therefore, the role of GPR120 agonists in dietary fat perception in humans was investigated herein. An emulsion prepared from vegetable oil had a stronger fatty orosensation, an orosensation similar to an oily mouth-coating sensed 5 - 10 s after tasting, than that prepared from mineral oil; however, the physical properties of both emulsions, such as viscosity, particle distribution, interfacial tension, contact angle, frictional load, and ζ-electric potential were similar. The potent GPR120 agonist, TUG-891 enhanced the fatty orosensation when added to the emulsion prepared from vegetable oil, but not to that from mineral oil. All GPR120 agonists tested enhanced the fatty orosensation when added to a low-fat food system whereas they did not evoke any fatty sensation in aqueous solution at the concentrations tested in food system, and sensory activity positively correlated with GPR120 activity. These results suggest that GPR120 agonists enhance the fatty orosensation in humans when added to vegetable oil or a low-fat food system, but do not evoke it by themselves.

Quantification of the kokumi peptide, γ-glutamyl-valyl-glycine, in cheese: Comparison between cheese made from cow and ewe milk.

Recent studies have shown that several types of cheese contain kokumi γ-glutamyl dipeptides, and the kokumi tripeptide, γ-glutamyl-valyl-glycine (γ-Glu-Val-Gly), is a component of various fermented foods. The quantification of γ-Glu-Val-Gly in various types of cheese was herein conducted by HPLC-tandem mass spectrometry followed by derivatization with 6-aminoquinoyl-N-hydroxysuccinimidyl-carbamate. The γ-Glu-Val-Gly concentrations were between 0.35 and 0.59 µg/g in cheese made from ewe milk, but were not detected in cheese made from cow milk. The amino acid sequences of major milk proteins showed that the ß-caseins of sheep had the Val-Gly sequence at the 9-10 position, whereas ß-caseins of cows contained a Pro-Gly sequence at the same position. The Val-Gly sequence was absent in other caseins of sheep and cattle. These results suggest that the different γ-Glu-Val-Gly concentrations present in cheese made from cow and ewe milk are due to differences in the amino acid sequences of caseins.

Association between soup consumption and obesity: A systematic review with meta-analysis.

This systematic review aimed to determine the correlation between soup consumption and obesity. The observational studies on the association of soup consumption to obesity-related parameters were screened by database search. From 1873 identified articles, 7 cross-sectional studies were included in the review. All studies indicated a significant inverse correlation between soup consumption and obesity. The meta-analysis of the studies of which outcome is odds ratio for obesity revealed that soup consumption is significantly related to lower odds ratio of obesity in combined data (n=45292, OR: 0.85, 95% CI: 0.79-0.91, p<0.0001), suggesting that soup consumption was inversely correlated with a risk of obesity.

Design, synthesis, and taste evaluation of a high-intensity umami-imparting oxazole-based compound.

Umami taste is imparted predominantly by monosodium glutamate (MSG) and 5'-ribonucleotides. Recently, several different classes of hydrophobic umami-imparting compounds, the structures of which are quite different from MSG, have been reported. To obtain a novel umami-imparting compound, N-cinnamoyl phenethylamine was chosen as the lead compound, and a rational structure-optimization study was conducted on the basis of the pharmacophore model of previously reported compounds. The extremely potent umami-imparting compound 2-[[[2-[(1E)-2-(1,3-benzodioxol-5-yl)ethenyl]-4-oxazolyle]methoxy]methyl]pyridine, which exhibits 27,000 times the umami taste of MSG, was found. Its terminal pyridine residue and linear structure are suggested to be responsible for its strong activity. The time taken to reach maximum taste intensity exhibited by it, as determined by the time-intensity method, is 22.0 s, whereas the maximum taste intensity of MSG occurs immediately. This distinct difference in the time-course taste profile may be due to the hydrophobicity and strong receptor affinity of the new compound.

Determination and quantification of kokumi peptide, γ-glutamyl-valyl-glycine, in brewed alcoholic beverages.

Recently, it has been demonstrated that kokumi substances such as glutathione are perceived through the calcium-sensing receptor (CaSR), and screening by CaSR assay and sensory evaluation has shown that γ-glutamyl-valyl-glycine (γ-Glu-Val-Gly) is a potent kokumi peptide. In this study, γ-Glu-Val-Gly contents in various brewed alcoholic beverages were investigated. Contents of γ-Glu-Val-Gly in four brands of wine, four brands of rice wine (sake) and eight brands of beer were analyzed by high performance liquid chromatography-tandem mass spectrometry followed by derivatization with 6-aminoquinoyl-N-hydroxysuccinimidyl-carbamate. The analyses indicated that γ-Glu-Val-Gly was present in all of eight beer samples at concentrations in the range of 0.08-0.18 mg/L, although the peptide was not detected in any wine or rice wine samples. These results suggest that amongst the brewed beverages tested, beer contains γ-Glu-Val-Gly, and that γ-Glu-Val-Gly is widely distributed in beer.

Determination and quantification of the kokumi peptide, γ-glutamyl-valyl-glycine, in commercial soy sauces.

Recent studies have demonstrated that kokumi substances, such as glutathione, are perceived through the calcium-sensing receptor (CaSR), and screening by CaSR assay and sensory evaluation has shown that γ-glutamyl-valyl-glycine (γ-Glu-Val-Gly) is a potent kokumi peptide. In this study, the contents of γ-Glu-Val-Gly in six commercial brands of dark-coloured soy sauces, two brands of light-coloured soy sauce, and one brand of white soy sauce, were investigated by high performance liquid chromatography-tandem mass spectrometry (LC/MS/MS), followed by derivatization with 6-aminoquinoyl-N-hydroxysuccinimidyl-carbamate (AQC). The analyses indicated that γ-Glu-Val-Gly was present in all investigated soy sauces at concentrations ranging from 0.15 to 0.61mg/dl, demonstrating that it is widely distributed in soy sauces.

Determination and quantification of γ-glutamyl-valyl-glycine in commercial fish sauces.

It was recently reported that kokumi substances such as glutathione are perceived through the calcium-sensing receptor (CaSR). In addition, screening by the CaSR assay and sensory evaluation revealed that γ-glutamyl-valyl-glycine (γ-Glu-Val-Gly) was a potent kokumi peptide. In this study, the quantities of γ-Glu-Val-Gly in various commercial fish sauces originating from Vietnam (Nuoc Mum), Thailand (Nampra), China (Yu-lu), Korea, Japan (Shottsuru and Ikanago-shoyu), and Italy (Garum) were investigated using a LC/MS/MS method followed by derivatization with 6-aminoquinoyl-N-hydroxysuccinimidyl-carbamate (AQC). The analyses revealed γ-Glu-Val-Gly at concentrations ranging from 0.04 to 1.26 mg/dL, indicating that γ-Glu-Val-Gly is widely distributed among various commercial fish sauces.

Kokumi substances, enhancers of basic tastes, induce responses in calcium-sensing receptor expressing taste cells.

Recently, we reported that calcium-sensing receptor (CaSR) is a receptor for kokumi substances, which enhance the intensities of salty, sweet and umami tastes. Furthermore, we found that several γ-glutamyl peptides, which are CaSR agonists, are kokumi substances. In this study, we elucidated the receptor cells for kokumi substances, and their physiological properties. For this purpose, we used Calcium Green-1 loaded mouse taste cells in lingual tissue slices and confocal microscopy. Kokumi substances, applied focally around taste pores, induced an increase in the intracellular Ca(2+) concentration ([Ca(2+)](i)) in a subset of taste cells. These responses were inhibited by pretreatment with the CaSR inhibitor, NPS2143. However, the kokumi substance-induced responses did not require extracellular Ca(2+). CaSR-expressing taste cells are a different subset of cells from the T1R3-expressing umami or sweet taste receptor cells. These observations indicate that CaSR-expressing taste cells are the primary detectors of kokumi substances, and that they are an independent population from the influenced basic taste receptor cells, at least in the case of sweet and umami.

Determination of γ-glutamyl-valyl-glycine in raw scallop and processed scallop products using high pressure liquid chromatography-tandem mass spectrometry.

The determination of the kokumi peptide, γ-glutamyl-valyl-glycine (γ-Glu-Val-Gly) in raw scallop and processed scallop products was carried out using high pressure liquid chromatography-tandem mass spectrometry (LC/MS/MS). The detection of γ-Glu-Val-Gly was achieved using a multiple reaction monitoring (MRM) method. The optimised condition enabled the precise determination of γ-Glu-Val-Gly. Raw scallop contained 0.08 µg/g γ-Glu-Val-Gly, and the γ-Glu-Val-Gly levels in processed scallop products, such as dried-scallop and scallop extract, were measured to be 0.64 and 0.77 µg/g, respectively. This is the first report to confirm the existence of γ-Glu-Val-Gly in foodstuff.

Frequency of soup intake is inversely associated with body mass index, waist circumference, and waist-to-hip ratio, but not with other metabolic risk factors in Japanese men.

Several previous studies have shown that the intake of soup negatively correlates with the body mass index (BMI), serum cholesterol and triacylglycerol levels, and blood pressure, suggesting that soup intake reduces metabolic risk. However, the correlation between soup intake and various metabolic risk factors has not been well-established. Especially, it has not been investigated in Asian countries. The aim of this study was to investigate the association of the frequency of soup intake and metabolic risk factors such as BMI, waist circumference, waist-to-hip ratio, serum cholesterol, serum triacylglycerol, blood glucose, and glycated hemoglobin. A cross-sectional study of 103 Japanese men aged 24 to 75 years was conducted. The intake of soup and other food was investigated by semi-quantitative food frequency questionnaires. The correlation between the frequency of soup intake and metabolic risk factors was analyzed by multiple regression analysis with a linear model. The median value of frequency of soup intake was 7.0 times per week. After adjusting for confounding factors such as age, energy intake, energy from alcohol intake, current smoking, and estimated energy expenditure, the frequency of soup intake was found to have a significant inverse association with BMI (P=0.040), waist circumference (P=0.024), and waist-to-hip ratio (P=0.001). However, no significant associations with other metabolic risk factors were found. Frequency of soup intake is negatively correlated with obesity-related physical parameters in Japanese men.

Consumption of bonito extract suppresses the decrease in cerebral blood flow in stroke-prone spontaneously hypertensive rats.

The effect of consuming bonito extract (BE) on cerebral blood flow was evaluated in stroke-prone spontaneously hypertensive rats (SHRSP), a cerebrovascular disease model. BE dissolved in drinking water was given to 5-week-old male SHRSP for 7 weeks. Tap water was given to the control group. At the age of 12 weeks, blood flow and vascular diameter were measured in the middle cerebral artery. Both cerebral blood flow and cerebral vessel width were greater in the BE group than in the control group. Also, stroke occurred in 7 (with death in 2) of the 8 animals in the control group but in none of the 6 animals in the BE group. To clarify its mechanism, the expressions of nitrogen oxide synthase (NOS) and the superoxide dismutase activity (SOD) in the brain were evaluated. NOS mRNA expression and SOD activity in the cerebrum were higher in the BE group. These results suggest that the consumption of BE suppresses the decrease of cerebral blood flow and reduces the risk of stroke to maintain vasorelaxation through the production of nitrogen oxide and suppression of active oxygen generation.

Frequency of soup intake and amount of dietary fiber intake are inversely associated with plasma leptin concentrations in Japanese adults.

Previous studies have shown that the intake of soup negatively correlates with the body mass index (BMI), suggesting that soup intake reduces the risk of obesity. In this study, to clarify the association of the intake of soup and various nutrients with plasma leptin concentration, a cross-sectional study on 504 Japanese adults aged 20-76 years (103 men and 401 women) was performed. The intake of soup and various nutrients was investigated by food frequency questionnaires. Plasma leptin concentration was measured in fasting blood by radioimmunoassay. The correlation was analyzed by multiple regression analysis. The average frequency of soup intake was 7.6 times/week. The average plasma leptin concentration was 7.76 ng/ml. After adjusting the confounding factors, the frequency of soup intake has a significant inverse association with plasma leptin concentration. Among the macronutrients, only dietary fiber intake negatively correlated with plasma leptin concentration after the adjustment for potential confounding factors. These results suggest that the intakes of soup and dietary fiber were negatively correlated with plasma leptin concentration in Japanese adults.

Ingestion of bonito extract ameliorates peripheral blood flow in mice loaded from over crowding stress.

Bonito extract (BE) has been shown to improve various fatigue-related symptoms. The possibility that the improvement of blood flow contributes to the improvement of fatigue-related symptoms has been reported. However, even though BE has been found to increase peripheral blood flow in humans, an understanding of its mechanisms has remained elusive. The purpose of the present study is to construct an animal model system with which the blood flow-increasing effects of BE can be examined. Using mice loaded with crowding stress, an attempt was made to reproduce the increases in peripheral blood flow observed in humans after a single administration of BE. In this study, the crowded-condition mice (20 mice/cage) showed significantly increased catecholamine levels (noradrenaline, adrenaline, and dopamine) in their circulating blood and a decreased rate of skin blood perfusion in comparison with the normal-condition mice (6 mice/cage). The rate of skin blood perfusion was significantly increased by BE in the crowded-condition mice in comparison with the controls, but not influenced by BE in the normal-condition mice. This suggests that BE expands the vascular diameter by affecting the constriction of vessels induced by catecholamines.

The anti-fatigue effects of the low-molecular-weight fraction of bonito extract in mice.

Bonito extract (BE), a hot-water extract of bonito muscle, has traditionally been considered as a folk remedy for fatigue. In this study we investigated the effects of BE on physical fatigue. BE was divided into, high, mid, and low-molecular-weight fractions (LMF), to explore the effectiveness of BE compounds. The swimming times to exhaustion of mice administered 0.86 g/kg BE and those administered 0.86 g/kg LMF were significantly longer than those of the vehicle-treated mice in a forced swimming model, indicating that BE possesses an anti-fatigue effect and that the LMF contributes to this effect. The LMF was also confirmed to aid the recovery of locomotor activity after physical fatigue in a forced walking model. We also examined respiratory gas levels and found that oxygen consumption and lipid oxidation were significantly greater in the group administered LMF than in the vehicle group, indicating that LMF promotes the utilization of fatty acids as an energy source. To elucidate why the mice administered LMF showed an anti-fatigue effect, we evaluated metabolic variables during exercise. The concentrations of nonesterified fatty acids and ketone bodies were higher, whereas serum and muscle lactic acid levels were lower in the mice administered LMF than in those in the vehicle group after the start of swimming. When the results were taken as a group they indicated that the effect of BE administration on improving endurance capacity was mediated, at least partly, by an increased utilization of lipids as a source of energy during exercise.

The Effect of the Dried-Bonito Broth on Blood Pressure, 8-Hydroxydeoxyguanosine (8-OHdG), an Oxidative Stress Marker, and Emotional States in Elderly Subjects.

Dried-bonito broth (DBB, katsuo-bushi dashi) is commonly used in Japanese cuisine, and is also used as a traditional remedy for recovery from fatigue and improvement of blood circulation. To clarify the effect of DBB on blood pressure, oxidative stress and emotional states, a randomized crossover human trial was performed. Twenty-seven elderly Japanese subjects ingested DBB or water for one month. Measurement of blood pressure and urinary 8-hydroxydeoxyguanosine (8-OHdG) and evaluation of emotional states were performed before and after the ingestion periods. The changes in systolic blood pressure (SBP) during DBB ingestion was significantly lower than that during water ingestion (p = 0.037). Urinary 8-OHdG significantly decreased during DBB ingestion (p = 0.0002). Evaluation of emotional states indicated that composure significantly improved during DBB ingestion (p = 0.034). These results suggest that the daily ingestion of DBB lower SBP, reduce urinary 8-OHdG and might improve emotional states in elderly subjects.

Effect of dried-bonito broth on mood states: a pooled analysis of four randomized controlled human trials.

Dried-bonito broth is commonly employed as a soup and sauce base in Japanese cuisine and is considered to be a nutritional supplement that promotes recovery from fatigue. Previous human trials have indicated that the ingestion of dried-bonito broth improves several mood states including fatigue, however, the effects in these studies have differed. The aim of this study was to clarify the effects of daily ingestion of dried-bonito broth on mood states by a pooled analysis of the randomized placebo-controlled trials. Five comparisons in four trials were selected for the pooled analysis (n = 159). The ingestion of dried-bonito broth significantly decreased the scores for fatigue (P = 0.032) and increased those for vigor (P = 0.027) compared to the placebo ingestion, suggesting that the dried-bonito broth improved fatigue felt in daily life. Furthermore, the ingestion of dried-bonito broth decreased the scores for tension-anxiety (P = 0.004) and confusion (P = 0.008) compared to the placebo ingestion. The ingestion of dried-bonito broth also significantly decreased the scores for total mood disturbance (P = 0.005). These results suggest that the ingestion of dried-bonito broth improves mood states especially fatigue, vigor, tension-anxiety and confusion.

Effect of dried-bonito broth intake on peripheral blood flow, mood, and oxidative stress marker in humans.

Dried-bonito broth (DBB) has been confirmed to improve various symptoms related to fatigue, but the reasons for this have remained unclear. Hypothesizing that DBB improves peripheral circulation together with mood states, we performed a randomized, double-blind, placebo-controlled crossover study in thirty-one healthy females. The subjects ingested DBB or a placebo for two weeks and changes in mood states after ingestion were investigated using the profile of mood states (POMS) questionnaire. The peripheral blood flow was also measured before and after ingestion of the test diet using a laser Doppler blood flow meter. The six mood factors and total mood disturbance score which reflect the mental states significantly improved, and a significant increase in peripheral blood flow was also found during DBB ingestion. As a result of correlation analysis between changes in each POMS score and changes in blood flow, it was suggested that a change in blood flow correlated with a change in some POMS factors and total mood disturbance. Based on these findings, we considered that blood flow may have increased in subjects whose mood states were markedly improved, suggesting that the improvement in mood states, including fatigue, was related to the increase in blood flow due to the improvement of peripheral circulation. To clarify whether DBB ingestion exhibits antioxidative activity, we investigated the urinary amounts of 8-hydroxy-2'-deoxyguanosine (8-OHdG) known as an oxidative stress marker and found that urinary excretion of the 8-OHdG for 24 h was significantly decreased during DBB ingestion. This study clarified that DBB ingestion improved mood states, increased peripheral blood flow, and decreased the oxidative stress marker.