Melanin Synthesis Inhibition Activity of Compounds Isolated from Bamboo Shoot Skin (Phyllostachys pubescens).

This study targets the evaluation of melanin synthesis inhibition activity of the bamboo shoot skin as agro-waste. The total methanolic extract of bamboo peel extract was evaluated for its skin protective effects via measuring its melanin inhibitory activity and its suppression activity on the expression of tyrosinase mRNA levels. Results showed that bamboo peel extract has a good ability for the inhibition of melanin synthesis so further studies were performed for the isolation of its constituents. Twelve compounds have been isolated from the shoot skin of Phyllostachys pubescens. Their structures were elucidated based on extensive spectroscopic methods. The melanin inhibition potential of the isolates was tested with their collagen-production-promoting activity for the determination of active principles. Results showed that Betulinic acid, tachioside, and 1,2-dilinolenin significantly suppressed melanin production per cell compared to control. Triacontanol, tricin, and (+)-lyoniresinol 9'-O-glucoside also tended to decrease melanin production per cell. These findings indicated that the skin of bamboo shoots, a significant agricultural waste, is a useful natural source for further research on its potential for aging problems such hyperpigmentation and cognitive function impairment.

Epb41l5 competes with Delta as a substrate for Mib1 to coordinate specification and differentiation of neurons.

We identified Erythrocyte membrane protein band 4.1-like 5 (Epb41l5) as a substrate for the E3 ubiquitin ligase Mind bomb 1 (Mib1), which is essential for activation of Notch signaling. Although loss of Epb41l5 does not significantly alter the pattern of neural progenitor cells (NPCs) specified as neurons at the neural plate stage, it delays their delamination and differentiation after neurulation when NPCs normally acquire organized apical junctional complexes (AJCs) in the zebrafish hindbrain. Delays in differentiation are reduced by knocking down N-cadherin, a manipulation expected to help destabilize adherens junctions (AJs). This suggested that delays in neuronal differentiation in epb41l5-deficient embryos are related to a previously described role for Epb41l5 in facilitating disassembly of cadherin-dependent AJCs. Mib1 ubiquitylates Epb41l5 to promote its degradation. DeltaD can compete with Epb41l5 to reduce Mib1-dependent Epb41l5 degradation. In this context, increasing the number of NPCs specified to become neurons, i.e. cells expressing high levels of DeltaD, stabilizes Epb41l5 in the embryo. Together, these observations suggest that relatively high levels of Delta stabilize Epb41l5 in NPCs specified as neurons. This, we suggest, helps coordinate NPC specification with Epb41l5-dependent delamination and differentiation as neurons.

Decreased stress-induced depression-like behavior in lactating rats is associated with changes in the hypothalamic-pituitary-adrenal axis, brain monoamines, and brain amino acid metabolism.

Depression-like behavior during lactation may relate to changes in the hypothalamic-pituitary-adrenal (HPA) axis, brain monoamines, and brain amino acid metabolism. This study investigated how the behavior, HPA axis activity, brain monoamines, and brain free amino acid metabolism of rats were changed by stress or lactation period. Rats were separated into four groups: (1) control lactating (n = 6), (2) stress lactating (n = 6), (3) control virgin (n = 7), and (4) stress virgin (n = 7) and restrained for 30 min a total of ten times (once every other day) from postnatal day (PND) 1. Depression-like behavior in the forced swimming test (FST) on PND 10 and concentration of corticosterone in plasma, as well as monoamines and L-amino acids including ß-alanine, γ-aminobutyric acid, cystathionine, 3-methyl-histidine and taurine in the prefrontal cortex and hypothalamus on PND 19 were measured. The plasma corticosterone concentration, measured just after restraint stress, was significantly higher in the stress groups, versus the control groups, but there were no significant differences between control and stress lactating groups. Depression-like behavior (immobility) in the FST was significantly lower in the lactating groups, versus the virgin groups. Stress enhanced dopamine and glutamate, and decreased threonine and glycine concentrations in the hypothalamus. In addition, 3-methoxy-4-hydroxyphenylglycol (MHPG), threonine and ornithine concentrations in the prefrontal cortex were significantly higher in the lactating groups compared with the virgin groups. Changes in plasma corticosterone concentration, monoamine, and amino acid metabolism may relate to stress-induced depression-like behavior in lactating rats. Lay summary This study revealed that reduced depression-like behavior in lactating, relative to virgin rats, was associated with changes in monoamine and amino acid metabolism in the prefrontal cortex of the brain. In addition, the effect of stress on monoamine and amino acid metabolism is prominently observed in the hypothalamus and may be related to neuroendocrine stress axis activity and secretion of corticosterone. This study suggested that stress-induced depression-like behavior may be associated with several changes in the stress axis, brain monoamines, and brain amino acid metabolism. These parameters were associated with attenuated depression-like behavior in lactating rats.

In ovoL-leucine administration stimulates lipid metabolisms in heat-exposed male, but not female, chicks to afford thermotolerance.

Thermal manipulation declined embryonic brain and liver concentrations of leucine (Leu). L-Leu in ovo injection afforded thermotolerance in male broiler chicks. This study aimed to examine the role of in ovo injection of L-Leu in metabolic functions, and differences between male and female broiler chicks in thermotolerance. L-Leu injection was performed in ovo on embryonic day (ED) 7 to reveal its role in metabolic activity in embryos and in post-hatch male and female broiler chicks under heat stress. To examine the metabolic activity of embryos, oxygen (O2) consumption, carbon dioxide (CO2) production, heat production and plasma metabolites were analyzed. Rectal temperature, food intake and plasma metabolites were also analyzed in heat-exposed (35 ± 1°C for 180min) male and female broiler chicks. It was found that O2 consumption, heat production, and plasma triacylglycerol (TG) and non-esterified fatty acid (NEFA) concentrations in ED 14 embryos were significantly increased by in ovo injection of L-Leu in comparison with the controls. Plasma glucose concentration was significantly increased in both male and female chicks under heat stress, but in ovo injection of L-Leu attenuated the increase in male chicks. In contrast, plasma TG, NEFA, and ketone body concentrations were significantly higher in male chicks injected in ovo with L-Leu, but not in similarly injected female chicks, compared with control chicks, all under heat stress. Rectal temperature and food intake were significantly lower in male, but not female, chicks under heat stress injected in ovo with L-Leu. In conclusion, in ovoL-Leu administration enhanced the prenatal metabolic rate and lipid metabolisms, which possibly appeared as sex-dependent fashion to facilitate thermotolerance in males. A reduction in heat production through lowered food intake in heat-exposed male, but not female chicks injected in ovo with L-Leu may help to afford thermotolerance in male broiler chicks under heat stress.

Gut microbiota of mice putatively modifies amino acid metabolism in the host brain.

Recently, it has been found that the gut microbiota influences functions of the host brain by affecting monoamine metabolism. The present study focused on the relationship between the gut microbiota and the brain amino acids. Specific pathogen-free (SPF) and germ-free (GF) mice were used as experimental models. Plasma and brain regions were sampled from mice at 7 and 16 weeks of age, and analysed for free d- and l-amino acids, which are believed to affect many physiological functions. At 7 weeks of age, plasma concentrations of d-aspartic acid (d-Asp), l-alanine (l-Ala), l-glutamine (l-Gln) and taurine were higher in SPF mice than in GF mice, but no differences were found at 16 weeks of age. Similar patterns were observed for the concentrations of l-Asp in striatum, cerebral cortex and hippocampus, and l-arginine (l-Arg), l-Ala and l-valine (l-Val) in striatum. In addition, the concentrations of l-Asp, d-Ala, l-histidine, l-isoleucine (l-Ile), l-leucine (l-Leu), l-phenylalanine and l-Val were significantly higher in plasma of SPF mice when compared with those of GF mice. The concentrations of l-Arg, l-Gln, l-Ile and l-Leu were significantly higher in SPF than in GF mice, but those of d-Asp, d-serine and l-serine were higher in some brain regions of GF mice than in those of SPF mice. In conclusion, the concentration of amino acids in the host brain seems to be dependent on presence of the gut microbiota. Amino acid metabolism in the host brain may be modified by manipulating microbiota communities.

l-Leucine acts as a potential agent in reducing body temperature at hatching and affords thermotolerance in broiler chicks.

Thermal manipulation (TM) of incubation temperature causes metabolic alterations and contributes to improving thermotolerance in chicks post hatching. However, there has been no report on amino acid metabolism during TM and the part it plays in thermotolerance. In this study, we therefore first analyzed free amino acid concentrations in the embryonic brain and liver during TM (38.6°C, 6h/d during embryonic day (ED) 10 to ED 18). It was found that leucine (Leu), phenylalanine and lysine were significantly decreased in the embryonic brain and liver. We then chose l-Leu and other branched-chain amino acids (l-isoleucine (L-Ile) and l-valine (l-Val)) for in ovo injection on ED 7 to reveal their roles in thermoregulation, growth, food intake and thermotolerance in chicks. It was found that in ovo injection of l-Leu, but not of l-Ileu or l-Val, caused a significant decline in body temperature at hatching and increased food intake and body weight gain in broiler chicks. Interestingly, in ovo injection of l-Leu resulted in the acquisition of thermotolerance under high ambient temperature (35±1°C for 180min) in comparison with the control thermoneutral temperature (28±1°C for 180min). These results indicate that the free amino acid concentrations during embryogenesis were altered by TM. l-Leu administration in eggs caused a reduction in body temperature at hatching, and afforded thermotolerance in heat-exposed young chicks, further suggesting that l-Leu may be one of the key metabolic factors involved in controlling body temperature in embryos, as well as in producing thermotolerance after hatching.

A single mutation in the acetylcholine receptor δ-subunit causes distinct effects in two types of neuromuscular synapses.

Mutations in AChR subunits, expressed as pentamers in neuromuscular junctions (NMJs), cause various types of congenital myasthenic syndromes. In AChR pentamers, the adult ε subunit gradually replaces the embryonic γ subunit as the animal develops. Because of this switch in subunit composition, mutations in specific subunits result in synaptic phenotypes that change with developmental age. However, a mutation in any AChR subunit is considered to affect the NMJs of all muscle fibers equally. Here, we report a zebrafish mutant of the AChR δ subunit that exhibits two distinct NMJ phenotypes specific to two muscle fiber types: slow or fast. Homozygous fish harboring a point mutation in the δ subunit form functional AChRs in slow muscles, whereas receptors in fast muscles are nonfunctional. To test the hypothesis that different subunit compositions in slow and fast muscles underlie distinct phenotypes, we examined the presence of ε/γ subunits in NMJs using specific antibodies. Both wild-type and mutant larvae lacked ε/γ subunits in slow muscle synapses. These findings in zebrafish suggest that some mutations in human congenital myasthenic syndromes may affect slow and fast muscle fibers differently.

Long-term consumption of dried bonito dashi (a traditional Japanese fish stock) reduces anxiety and modifies central amino acid levels in rats.

Dried bonito dashi, a traditional Japanese fish stock, enhances palatability of various dishes because of its specific flavor. Daily intake of dashi has also been shown to improve mood status such as tension-anxiety in humans. This study aimed at investigating beneficial effects of dashi ingestion on anxiety/depression-like behaviors and changes in amino acid levels in the brain and plasma in rats. Male Wistar rats were given either dried bonito dashi or water for long-term (29 days; Experiment 1) or single oral administration (Experiment 2). Anxiety and depression-like behaviors were tested using the open field and forced swimming tests, respectively. Concentrations of amino acids were measured in the hippocampus, hypothalamus, cerebellum, and jugular vein. During the long-term (29 days) consumption, rats given 2% dashi frequently entered the center zone and spent more time compared with the water controls in the open field test. However, the dashi was ineffective on depression-like behavior. In the hippocampus, concentrations of hydroxyproline, anserine, and valine were increased by dashi while those of asparagine and phenylalanine were decreased. In the hypothalamus, the methionine concentration was decreased. In a single oral administration experiment, the dashi (1%, 2% or 10%) showed no effects on behaviors. Significance was observed only in the concentrations of α-aminoadipic acid, cystathionine, and ornithine in the hippocampus. Dried bonito dashi is a functional food having anxiolytic-like effects. Daily ingestion of the dashi, even at lower concentrations found in the cuisine, reduces anxiety and alters amino acid levels in the brain.

Neck circumference has possibility as a predictor for metabolic syndrome in postmenopausal women.

Subcutaneous fat depots play an important role in regulating metabolic profile in Japanese postmenopausal women. We investigated the possibility of neck circumference (NC) as a surrogate marker for metabolic disease risk estimates in Japanese postmenopausal women. We examined the association of NC with several markers of insulin resistance, lipid metabolism and atherosclerosis in 64 healthy postmenopausal women aged 63.6 ± 7.1 years in community-based samples in Japan. As a result, NC was significantly associated with indices of whole body obesity and visceral fat accumulation, such as body mass index (BMI) and Waist circumference (WC). In the analysis of biomarkers for insulin resistance, NC was positively correlated to HbA1c, homeostasis model assessment ratio (HOMA-R) and leptin. In addition, an increase in triglycerides (TG) and a decrease in HDL-cholesterol (HDL-C) were also associated with NC. Interestingly, NC was also associated with atherosclerosis-related indices. The measurement of NC is an easy, inexpensive and reproducible method for assessment of obesity, and a possible predictor to identify the risk for future metabolic diseases in Japanese postmenopausal women.

Thermographic study on the preservability of heat effects of footbath with salt.

Infrared thermography provides a non-invasive and dynamic measure of heat. The thermal preservability effects of a salt footbath were evaluated by the infrared thermography technique. The subjects were 23 healthy college students. Feet were soaked for 10 min in a 40-42 degrees C normal footbath. Room temperature was set at 26.5-28 degrees C. At the same time on another day within 3 days of the normal footbath experiment, the same feet were soaked for 10 min in a 40-42 degrees C salt footbath. We measured blood pressure, heart rate and temperatures of the feet, second toes, hands and middle fingers, just before and after immersion and at 10-min intervals thereafter. Mean blood pressure changes showed no difference between the normal and the salt footbath. Mean heart rate changes were higher during the normal footbath than at 0, 15 and 20 min during the salt footbath, respectively (p < 0.05). Mean thermal preservability of the feet tended to be lower after the normal footbath than at 20 and 30 min, respectively, after the salt footbath, but these differences did not reach a statistical significance. Mean thermal preservability of the hands and middle fingers was significantly lower after the normal footbath than at 20 and 30 min, respectively, after the salt footbath (p < 0.05). The results suggest that stimulation by a salt footbath affects surface skin temperature, and that stimulation aimed at increasing skin thermal preservability shows a significant difference between normal and salt footbaths.

Acetylcholine receptors enable the transport of rapsyn from the Golgi complex to the plasma membrane.

The accumulation of acetylcholine receptors (AChRs) at nerve terminals is critical for signal transmission at the neuromuscular junction, and rapsyn is essential for this process. Previous studies suggest that AChRs might direct rapsyn self-clusters to the synapse. In vivo experiments with fluorescently tagged AChR or rapsyn in zebrafish larvae revealed that rapsyn self-clusters separate from AChRs did not exist before synapse formation. Examination of rapsyn in the AChR-less mutant sofa potato revealed that rapsyn in the absence of AChR was localized in the Golgi complex. Expression of muscle-type AChR in sofa potato restored synaptic clustering of rapsyn, while neuronal type AChR had no effect. To determine whether this requirement of protein interaction is reciprocal, we examined the mutant twitch once, which has a missense mutation in rapsyn. While the AChRs distributed nonsynaptically on the plasma membrane in twitch once, mutant rapsyn was retained in the Golgi complex. We conclude that AChRs enable the transport of rapsyn from the Golgi complex to the plasma membrane through a molecule-specific interaction.

Changes in bone mineral density and metabolism in women: evaluation of bodily characteristics, bone metabolic markers and bone mineral density.

The relationship of bone mineral density (BMD) and bone metabolic markers in women is an interesting field of research. In this study, we aimed to clarify the relationship of body weight, bone metabolic markers and BMD. The subjects were 72 women. The levels of serum bone-specific alkaline phosphatase (BAP), serum type I collagen-cross-linked peptide (s-NTx) and urinary deoxypyridinoline (u-DPD) were measured. The associations between dependent variables (BMD changes/1 or 4 years in the lumbar spine and femoral neck) and explanatory variables (body weight changes/1 or 4 years, the levels of BAP, s-NTx, u-DPD) were evaluated using multiple regression analysis. Changes in the lumbar spine BMD were significantly correlated with changes in height over a year, and those of the femoral neck were significantly correlated with changes in weight over a year. Changes in the lumbar spine BMD over 4 years were significantly correlated with age, BAP and the changes of weight over 4 years. Changes in the femoral neck BMD over 4 years were significantly correlated with the changes in weight for 4 years. These results suggest that BMD changes of different bones correlate with different explanatory variables and that, to predict BMD changes from bone metabolic markers in women, it is necessary to measure BAP levels.

Differential energy expenditure is involved in the difference in activity levels between the Djungarian hamster (Phodopus sungorus) and the Roborovskii hamster (P. roborovskii).

The Djungarian hamster (Phodopus sungorus) shows calm behavior, while the Roborovskii hamster (P. roborovskii) exhibits hyperactivity. Even though they belong to the same genus, Phodopus, these two species are quite different. The current study investigated the relationship between energy expenditure and the markedly different levels of activity shown by these hamsters. Roborovskii hamsters showed significantly higher energy expenditure than Djungarian hamsters under both feeding and fasting conditions during darkness. Roborovskii hamsters showed a repeated increase and decrease in energy expenditure under the feeding condition; however, this changed under the fasting condition, during which the repeated increase and decrease in energy expenditure corresponded to the repeated active and sleeping conditions. Djungarian hamsters had a tendency to keep their energy expenditure constant during the fasting condition, while Roborovskii hamsters moved around a lot to find food. The respiratory quotient (RQ) values in Djungarian hamsters were relatively constant. However, Roborovskii hamsters showed a wide variation in RQ. In particular, the RQ value declined immediately before a dark phase commenced, indicating a switchover from the utilization of glucose to that of lipids as a substrate for energy production. In conclusion, Djungarian hamsters and Roborovskii hamsters showed different behavioral patterns that were related to differences in energy metabolism.

Brazilian Propolis Intake Decreases Body Fat Mass and Oxidative Stress in Community-Dwelling Elderly Females: A Randomized Placebo-Controlled Trial.

This study aimed to investigate the effects of Brazilian propolis on body fat mass and levels of adiponectin and reactive oxygen species among community-dwelling elderly females. This was a double-blind randomized placebo-controlled trial. Altogether, 78 females aged 66-84 years were randomly assigned to the propolis (PRO; n = 39) or placebo (PLA; n = 39) group. For 12 weeks, the PRO group were given three capsules containing 227 mg of propolis twice a day. Meanwhile, the PLA group were given daily placebo capsules. Of 78 participants, 53 (PLA group: n = 28, PRO group: n = 25) completed the study. Although no changes were observed in absolute or relative fat mass in the PLA group, they showed a significant decline in the PRO group. The level of serum adiponectin in the PLA group did not change, although that of the PRO group significantly increased. The level of d-ROMs in the PLA group significantly increased, whereas that of the PRO group significantly decreased. The serum SOD activity in the PLA group significantly decreased, whereas that of the PRO group tended to increase. These results suggest that propolis supplementation may decrease body fat mass and oxidative stress among community-dwelling elderly females.

Factors predicting bone mineral density (BMD) changes in young women over a one-year study:changes in body weight and bone metabolic markers during the menstrual cycle and their effects on BMD.

Currently, 26% of Japanese women in their twenties are under weight, and therefore at risk of developing various metabolic abnormalities due to an inadequate nutrient intake, which in turn affects the acquisition of a peak bone mineral density (BMD). In this study, we aimed to clarify the effects of menstrual cycle-related changes in body weight and bone metabolic marker levels on the BMD changes. The subjects were 42 women (19.6 ± 0.8 years). The levels of osteocalcin (OC), BAP, s-NTx, u-DPD, and E2 in the menstrual and ovulatory phases were measured. The associations between dependent variables (BMD changes/year in the lumbar spine, femur, femoral neck) and explanatory variables (body weight changes/year, the levels of OC, BAP, s-NTx, u-DPD) were evaluated using multiple regression analysis. Analysis of the correlations between the changes in bone metabolic markers and changes in BMD showed a correlation between the OC level in the menstrual phase and changes in the BMD of the entire femur, suggesting that a high OC level protects against BMD reduction, probably by promoting osteoblast activity, and that bone formation activity suppresses the decrease in BMD. These results suggest that, to predict BMD changes from bone metabolic markers in young women, it is necessary to measure OC levels in the menstrual phase.

Longitudinal study on physical fitness parameters influencing bone mineral density reduction in middle-aged and elderly women: bone mineral density in the lumbar spine, femoral neck, and femur.

The prolongation of the average life span of women has been associated with the rapidly aging society. However, serious problem have arisen as a result, such as an increase in the number of bed-ridden elderly patients with osteoporosis-associated femoral neck fracture. As preventive measures against osteoporosis for middle-aged to elderly women, 10,000 steps per day and intense exercise have been reported to inhibit bone mineral density (BMD) reduction. However, only a few studies have concretely reported on the type of physical fitness that is effective for BMD in particular parts of the body. In this study, a one-year longitudinal survey was performed involving generally healthy postmenopausal women to investigate physical fitness parameters influencing BMD in the lumbar spine, femoral neck, and femur. The subjects were 38 female residents of M City, aged 49-73 years. As physical fitness parameters, sit-ups, anteflexion in a sitting position, grip strength, mean amount of exercise (kcal), and area of outer body sway on standing straight with the eyes closed (m2) were measured. The BMD was measured in the lumbar spine (L2-L4), femoral neck, and femur. Logistic regression analysis was performed regarding the physical fitness parameters as explanatory variables and groups with and without BMD reduction over one year as those with and without risk as dependent variables. The number of sit-ups (odds ratio: 0.76, 95% CI: 0.61-0.96, p=0.022) was a preventive factor against BMD reduction of the lumbar spine, and ante flexion in a sitting position was a preventive factor against BMD reduction of the femoral neck (odds ratio: 0.88, 95% CI: 0.78-0.99, p=0.029). Regarding BMD reduction of the femur, the area of outer body sway on standing straight with the eyes closed tended to be not significant to the risk. It is suggested that physical fitness and local muscle strength are associated with BMD reduction in the lumbar spine, femoral neck, and femur.

Dysregulated metabolism and behaviors by disrupting gut microbiota in prenatal and neonatal mice.

The live microbiota ecosystem in the intestine plays a critical role in maintaining the normal physiological and psychological functions in both animals and human beings. However, the chronic effect of microbiota disturbances during prenatal and neonatal developing periods on animal's health remains less studied. In the current study, pregnant ICR mice were fed with an antibiotic diet (7-g nebacitin [bacitracin-neomycin sulphate 2:1]/kg standard diet) from day 14 of conception, and their offspring were provided with the same diet till the termination of the experiments. Dams treated with antibiotics showed increased body weight along with enlarged gut. Antibiotic-treated offspring revealed decreased bodyweight, increased food, water, and sucrose intake. Administration of antibiotics affected corticosterone responsivity to acute 20 min restraint challenge in male pups. In behavior tests, female pups showed decreased movement in open field while male pups revealed decreased latency to open arms in elevated plus maze test and immobility time in tail suspension test. Together, these results suggested that early antibiotic exposure may impact on the food intake, body weight gain, and emotional behavior regulation in mice.

Altered Fecal Microbiotas and Organic Acid Concentrations Indicate Possible Gut Dysbiosis in University Rugby Players: An Observational Study.

Gut eubiosis is essential for the host's health. In athletes, the gut microbiota can be altered by several factors, including diets. While eubiotic gut microbiota in elite rugby players has been reported, our survey found that university rugby players suffered from loose stools and frequent urgency to defecate. To establish the causes of the condition, the microbiota and the concentrations of organic acids in fecal samples of university male rugby players (URP) were analyzed and compared with those of age-matching, non-rugby playing males (control). Body mass indices were significantly (p < 0.05) different between groups. Chao1 index was significant (p < 0.05) lower in URP than in control. The relative abundances of phyla Firmicutes and Bacteroidetes were significantly (p < 0.05) higher and lower, respectively, in URP than in control. Potential pathobiont genera Collinsella, Enterobacter, and Haemophilus were significantly (p < 0.05) abundant, whereas beneficial Akkermansia was lower (p < 0.05) in URP than in control. Succinate, a potential causative of gut inflammation, was five-fold higher in URP than in controls. Our findings all but confirmed that the dysbiotic status of gut in URP.

Effects of Lemon Beverage Containing Citric Acid with Calcium Supplementation on Bone Metabolism and Mineral Density in Postmenopausal Women: Double-Blind 11-Month Intervention Study.

A critical factor for preventing osteoporosis after menopause is attenuation of the accelerated turnover rate of bone metabolism. The present randomized controlled study was conducted to clarify the effects of a lemon beverage with calcium (Ca) supplementation that makes use of the chelating action of citric acid. Comprehensive evaluations of bone were performed by assessments of bone mineral density (BMD) and biomarkers related to bone turnover. Seventy-nine postmenopausal women were enrolled and asked to participate in an 11-month continuous intake of the test beverages. The subjects were divided into three groups: those who consumed a lemon beverage containing citric acid with Ca supplementation (LECA group), those who consumed a lemon beverage containing citric acid without Ca supplementation (LE group), and those who consumed no test beverage (control group). Using a double-blind protocol, subjects in the LECA and LE groups consumed one bottle containing 290 mL of the test beverage each day. The ratio of change in BMD after 11 months was significantly higher in the LECA group as compared to the control and LE groups. The LECA group also showed significant decreases in concentrations of tartrate-resistant acid phosphatase 5b (TRACP-5b), a bone resorption marker, and bone alkaline phosphatase (BAP) as compared to the other groups, as well as a significant decrease in concentration of osteocalcin (OC), a bone formation marker, as compared to the LE group. Based on our findings, we speculated that bone resorption and bone formation in postmenopausal women might be suppressed along with an increase in Ca resorption caused by chelation of citric acid in association with continuous ingestion of a Ca-supplemented lemon beverage containing citric acid, resulting in suppression of high bone metabolic turnover. In addition, the results provide information regarding BMD maintenance in the bones of the trunk, including the lumbar spine and proximal femur.

Dried Watermelon Rind Mash Diet Increases Plasma l-Citrulline Level in Chicks.

Heat stress is an increasing concern in poultry industry as it can cause a rise in the body temperature of chickens. Recently, we reported that l-citrulline (l-Cit) is a potential hypothermic agent that could improve thermotolerance in chicks. However, synthetic l-Cit has not yet been approved for inclusion in animal diets. l-Cit was first isolated from watermelon. Watermelon rind (WR), an agricultural waste product, contains more l-Cit than the flesh of the fruit. In the current study, the chemical composition and l-Cit content of WR dried powder (WRP) were determined. WRP was mixed with water at a ratio of 4:5 (wt/v) to make WRP mash, and then mixed with a commercial starter diet to prepare a 9% WRP mash diet. The WRP mash diet was fed to 3- to 15-day-old chicks and daily food intake, body weight, and changes in rectal temperature were measured. At the end of the experiment, blood was collected from the chicks to analyze plasma l-Cit and other free amino acids. The chemical analysis of WRP revealed a variety of components including 19.1% crude protein. l-Cit was the most abundant free amino acid in WRP (3.18 mg/g). Chronic supplementation of the WRP mash diet significantly increased compensatory food intake, plasma l-Cit, l-ornithine, and l-tyrosine in chicks. WRP mash diet did not affect the body temperature of the chicks. In conclusion, WRP mash diet supplementation increased plasma l-Cit concentration in chicks. The increase in plasma l-Cit concentrations suggest that WR could be used as a natural source of l-Cit in chicks to ameliorate the adverse effects of heat stress.