Age-related decline of motor unit firing rate in community-dwelling healthy older adults and daily ingestion of fish protein of Alaska pollack: a randomised trial.

Neuromuscular properties decline with ageing and low-level physical activities. Alaska pollack protein (APP) is reportedly effective for improving skeletal muscular functions, even if the amount is small and exercise is not conducted. However, it is unclear whether APP intake without an exercise program affects neuromuscular dysfunction in community-dwelling adults conducting normal, everyday activities. This study aimed to investigate changes in motor unit firing properties by adding APP to daily meals in community-dwelling older adults. They were divided into an APP group or a placebo control (CON) group for randomised, double-blind treatment. Participants in APP (n = 15) and CON (n = 17) groups ingested 4.5 g/d APP and whey protein, respectively, added to daily meals for 3 months without a specific exercise intervention. High-density surface electromyography of the vastus lateralis to assess the individual motor unit firing rate during 70% of maximum voluntary ramp-up contraction (MVC), maximum knee extensor strength, muscle thickness, echo intensity, and a physical function test were evaluated at the baseline, as well as 1.5 and 3 months after the intervention. While muscle strength, muscle thickness, and motor function were not changed, the motor unit firing rate at 50%-70% of MVC showed a significant decline at 1.5 and 3 months in the CON group, but this decline was not observed in the APP group. These findings suggest that neuromuscular properties show a detectable decline on performing daily activities without a specific exercise intervention, but such a decline was not observed in the APP intake group.

Chronic Intake of a Meal Including Alaska Pollack Protein Increases Skeletal Muscle Mass and Strength in Healthy Older Women: A Double-Blind Randomized Controlled Trial.

BACKGROUND: In animal studies, a meal containing Alaska pollack protein (APP) induces fast-twitch muscle hypertrophy. To our knowledge, no interventional studies have examined the benefits of APP intake on muscle mass and muscle weakness and the prevention of sarcopenia in older individuals. OBJECTIVES: We evaluated the effects of APP intake on skeletal muscle mass, muscle strength, and physical performance among healthy community-dwelling older Japanese women. METHODS: In this double-blind randomized controlled trial, healthy women ≥ 65 y old were allocated to an APP or whey protein control (CON) group. Participants ingested test protein meals (5.0-5.1 g protein/serving) daily for 24 wk. Between-group differences in the change of skeletal muscle mass index (SMI) as the primary outcome and muscle strength as a secondary outcome were tested using multifrequency BIA and a handheld dynamometer, respectively, at baseline, and 4, 12, and 24 wk. The mean changes in the measured primary and secondary outcome variables from baseline to 4, 12, and 24 wk were compared using unpaired t tests. RESULTS: There were no between-group differences in nutritional status, food intake, or total energy and protein intakes at baseline, 12 wk, or 24 wk. The change in SMI was 0.12 kg/m2 (95% CI: 0.01, 0.23 kg/m2) and 0.11 kg/m2 (95% CI: 0.03, 0.19 kg/m2) greater in the APP group than in the CON group at 12 wk and 24 wk (P ≤ 0.03) and knee extension strength was 0.07 Nm/kg BW (95% CI: 0.02, 0.12 Nm/kg BW) and 0.05 Nm/kg BW (95% CI: 0.00, 0.09 Nm/kg BW) higher in the APP group than in the CON group at these times (P ≤ 0.015), respectively. The groups did not differ at 4 wk. CONCLUSIONS: Daily intake of a meal containing APP compared with whey protein increases skeletal muscle mass and lower-extremity muscle strength in healthy older women, suggesting that an APP-containing meal may be useful in the prevention of sarcopenia in this group.This trial was registered at as UMIN000035718.

Essential biphasic role for JAK3 catalytic activity in IL-2 receptor signaling.

To drive lymphocyte proliferation and differentiation, common γ-chain (γc) cytokine receptors require hours to days of sustained stimulation. JAK1 and JAK3 kinases are found together in all γc-receptor complexes, but how their respective catalytic activities contribute to signaling over time is not known. Here we dissect the temporal requirements for JAK3 kinase activity with a selective covalent inhibitor (JAK3i). By monitoring phosphorylation of the transcription factor STAT5 over 20 h in CD4(+) T cells stimulated with interleukin 2 (IL-2), we document a second wave of signaling that is much more sensitive to JAK3i than the first wave. Selective inhibition of this second wave is sufficient to block cyclin expression and entry to S phase. An inhibitor-resistant JAK3 mutant (C905S) rescued all effects of JAK3i in isolated T cells and in mice. Our chemical genetic toolkit elucidates a biphasic requirement for JAK3 kinase activity in IL-2-driven T cell proliferation and will find broad utility in studies of γc-receptor signaling.

Covalent docking of large libraries for the discovery of chemical probes.

Chemical probes that form a covalent bond with a protein target often show enhanced selectivity, potency and utility for biological studies. Despite these advantages, protein-reactive compounds are usually avoided in high-throughput screening campaigns. Here we describe a general method (DOCKovalent) for screening large virtual libraries of electrophilic small molecules. We apply this method prospectively to discover reversible covalent fragments that target distinct protein nucleophiles, including the catalytic serine of AmpC ß-lactamase and noncatalytic cysteines in RSK2, MSK1 and JAK3 kinases. We identify submicromolar to low-nanomolar hits with high ligand efficiency, cellular activity and selectivity, including what are to our knowledge the first reported reversible covalent inhibitors of JAK3. Crystal structures of inhibitor complexes with AmpC and RSK2 confirm the docking predictions and guide further optimization. As covalent virtual screening may have broad utility for the rapid discovery of chemical probes, we have made the method freely available through an automated web server (http://covalent.docking.org/).

Effects of Alaska Pollack Protein Ingestion on Neuromuscular Adaptation in Young Healthy Adults: A Randomized, Placebo-Controlled Trial.

Alaska pollack protein (APP), has been reported as a protein source that can enhance muscle hypertrophy more than other protein sources in animal studies. This study aimed to examine the effects of APP ingestion on muscle quantity and quality in young adults. Fifty-five young college students were assigned to two groups: APP and placebo (whey protein: WP) groups, and instructed to ingest 4.5 g of each protein in addition to daily meals, and to maintain their usual daily physical activities for 3 mo. Twenty-one and 23 students completed the intervention and were analyzed in APP and WP groups, respectively. The maximum knee extension torque significantly increased in both groups during the intervention. The motor unit discharge rate, which is an indicator of activation, for a given force level significantly decreased in both groups during the intervention, but its decrease in the APP group was significantly greater than in the WP group. Echo intensity of the vastus lateralis evaluated by ultrasound images significantly decreased in both groups. The muscle thickness and skeletal muscle mass did not change. Small amount of additional APP intake induces greater effects on neural activation than WP, suggesting the greater neural economy of generation of force.

Effects of Immature Soybeans on Lipid Metabolism in Rats Fed Normal and High-Fat High-Fructose NAFLD Diets.

Edamame and chamame (brown edamame) are immature soybeans that contain abundant protein, vitamins, and dietary fiber. Edamame has a different nutrient composition compared with mature soybeans, as evidenced by its higher content of vitamins and folic acid. Recent studies have shown that proteins in mature soybeans, such as ß-conglycinin, and dietary fiber influence the metabolism of lipids such as triglyceride and cholesterol. However, the effect of edamame on lipid metabolism has not been reported. In the present study, we investigated whether oral consumption of edamame or chamame affects lipid metabolism. We focused on the analysis of hepatic lipids, serum cholesterol, and fecal lipids in rats after feeding them a normal or non-alcoholic fatty liver disease (NAFLD) diet containing edamame or chamame for 6 wk. We found that the consumption of dietary edamame and chamame significantly lowered hepatic triglyceride and serum cholesterol levels compared with those in the control groups fed with normal and NAFLD diets. Intake of edamame and chamame improved the NAFLD-like fatty liver. Finally, intake of edamame and chamame resulted in a significantly higher amount of fecal lipids and bile acid than in the control, suggesting the stimulation of lipid excretion. Together, dietary edamame and chamame decrease hepatic and serum cholesterol by promoting lipid excretion, not by the regulation of hepatic gene expression, leading to the improvement of lipid metabolism in both normal and NAFLD model rats.

Lactococcus lactis subsp. cremoris YRC3780 modifies function of mesenteric lymph node dendritic cells to modulate the balance of T cell differentiation inducing regulatory T cells.

Introduction: The intestinal immune system plays a pivotal role in the induction of immune responses against food. In the case of T cell response, dendritic cells (DCs) are especially important. However, the regulation of immune responses to food by intestinal DCs has been poorly described. In this study, we analyzed the effect of Lactococcus lactis subsp. cremoris YRC3780, a lactic acid bacterial strain isolated from kefir, a traditional fermented milk product, on the immune responses induced by antigen presentation by intestinal DCs to T cells as well as the mechanism of action of these immunomodulatory effects. It has been shown that L. cremoris YRC3780 ameliorates the symptoms of pollinosis in both animal and human studies. Methods: CD11c+ cells from mesenteric lymph nodes (MLNs) of BALB/c mice were cultured as MLN DCs with L. cremoris YRC3780 and expression of genes inducing regulatory T cells (Tregs) was examined by qPCR. In addition, MLN DCs were cocultured with CD4+ T cells from DO11.10 transgenic mice expressing an ovalbumin (OVA)-specific TCR and the OVA antigen peptide and L. cremoris YRC3780. Induction of Tregs was examined by flow cytometry, gene expression was analyzed by DNA microarray and qPCR, and the production of cytokines was measured by ELISA. MLN DCs from TLR2-deficient mice and components of L. cremoris YRC3780 were used to examine the recognition of YRC3780 by MLN DCs. Results: L. cremoris YRC3780 enhanced the expression of genes involved in Treg induction in MLN DCs and induced Foxp3+CD4+T cells in an MLN DC and CD4+ T-cell co-culture system. The effect on MLN DCs was likely mediated by receptors other than TLR2. Together with microarray analyses of CD4+ T cell gene expression and cytokine ELISA, it was demonstrated that L. cremoris YRC3780 promoted the induction of Th1 and Tregs, and regulated the balance of Th1/Th2 and Treg/Th17 cells involving multiple genes via the antigen-presentation of MLN DCs. Discussion: Our findings provide insights into the modulation of intestinal immune responses mediated by DCs and the antiallergic effects of lactic acid bacteria.

Comparison of myocardial perfusion imaging between the new high-speed gamma camera and the standard anger camera.

BACKGROUND: Cadmium-zinc-telluride (CZT) solid-state detectors have been recently introduced into the field of myocardial perfusion imaging. The aim of this study was to prospectively compare the diagnostic performance of the CZT high-speed gamma camera (Discovery NM 530c) with that of the standard 3-head gamma camera in the same group of patients. METHODS AND RESULTS: The study group consisted of 150 consecutive patients who underwent a 1-day stress-rest (99m)Tc-sestamibi or tetrofosmin imaging protocol. Image acquisition was performed first on a standard gamma camera with a 15-min scan time each for stress and for rest. All scans were immediately repeated on a CZT camera with a 5-min scan time for stress and a 3-min scan time for rest, using list mode. The correlations between the CZT camera and the standard camera for perfusion and function analyses were strong within narrow Bland-Altman limits of agreement. Using list mode analysis, image quality for stress was rated as good or excellent in 97% of the 3-min scans, and in 100% of the ≥4-min scans. For CZT scans at rest, similarly, image quality was rated as good or excellent in 94% of the 1-min scans, and in 100% of the ≥2-min scans. CONCLUSIONS: The novel CZT camera provides excellent image quality, which is equivalent to standard myocardial single-photon emission computed tomography, despite a short scan time of less than half of the standard time.

A preparation of cow's late colostrum fraction containing αs1-casein promoted the proliferation of cultured rat intestinal IEC-6 epithelial cells.

Colostrum is a complex mixture of bioactives that promotes neonate growth. Recently, we have found by in vivo study that skimmed, sterilized, and concentrated bovine late colostrum (SCBLC), obtained from a Holstein herd on days 6-7 after parturition, had an ability to maintain intestinal integrity. In the present study we investigated effects of SCBLC on rat intestinal IEC-6 cell proliferation in vitro. A fraction containing αs1-casein was found to have a robust stimulation effect as compared to other protein fractions from SCBLC and even the αs1-casein fraction from milk from other Holstein herds. Furthermore, the SCBLC αs1-casein molecule demonstrated not only slightly slower mobility on both SDS- and native-PAGE than other bovine milk αs1-caseins, but also a peculiar conformation reminiscent of moltenglobule in the circular dichroism spectrum. These findings may be of relevant to the competence of SCBLC to preserve intestinal integrity.

Effects of Lactococcus lactis subsp. cremoris YRC3780 daily intake on the HPA axis response to acute psychological stress in healthy Japanese men.

BACKGROUND: Lactococcus lactis subsp. cremoris (YRC3780), which is isolated from kefir, has been associated with anti-allergic effects in humans. However, it remains unknown whether daily intake of YRC3780 attenuates the response to psychological stress in humans in parallel with changes to the gut microbiome. We examined the fundamental role of YRC3780 in the gut microbiome, stress response, sleep, and mental health in humans. METHODS: Effects of daily intake of YRC3780 on the hypothalamic-pituitary-adrenal (HPA) axis response to acute psychological stress were investigated in a double-blind, placebo-controlled clinical trial involving 27 healthy young men (mean age and body mass index: 23.5 years and 21.5 kg/m2) who were randomly assigned to placebo (n = 13) or YRC3780 (n = 14) groups. The HPA axis response to acute psychological stress, the diurnal rhythm of HPA axis activity, and gut microbiome were assessed and compared between the two groups. RESULTS: The results showed that daily intake of YRC3780 significantly lowered morning salivary cortisol levels compared with placebo. In addition, salivary cortisol levels following a social stress test significantly decreased +40 min after beginning the TSST in the YRC3780-treated group compared to placebo. There were no significant differences between the two groups in terms of actigraphy-based sleep quality, but the subjective sleep quality and mental health were significantly improved in the YRC3780-treated group compared to placebo. CONCLUSIONS: Our study suggests that daily intake of YRC3780 improves the HPA axis response to acute psychological stress, which might be associated with a decrease in morning cortisol levels.

Suppressive Effect of Lactococcus lactis subsp. cremoris YRC3780 on a Murine Model of Japanese Cedar Pollinosis.

Accumulating evidence suggests that Lactococcus lactis subsp. cremoris YRC3780 isolated from kefir has the potential to alleviate allergic responses. Herein, we investigated the effect of YRC3780 on a murine model of Japanese cedar pollinosis (JCP). BALB/c mice immunized with cedar pollen extract (CPE) exhibited an increase in serum immunoglobulin E and developed nasal inflammatory responses including sneezing, nasal hyperresponsiveness, and nasal eosinophil accumulation upon intranasal allergen challenge. These responses were suppressed by the oral administration of YRC3780, although the effects on CPE-induced sneezing response and eosinophil infiltration were not statistically significant. Total fecal microbiota diversity was not affected by allergen immunization and challenge or by YRC3780 administration. However, the abundances of Bifidobacteriales, Veillonellaceae, Lactococcus, and Lactococcus lactis were larger and that of Bacteroides was smaller in YRC3780-treated mice compared with those in CPE-challenged and YRC3780-untreated mice. Our findings suggest the usefulness of YRC3780 for alleviating JCP.

Fish Protein Promotes Skeletal Muscle Hypertrophy via the Akt/mTOR Signaling Pathways.

Skeletal muscle is the largest organ in the body and has a broad range of plasticity, undergoing atrophy in response to aging or disease and hypertrophy in response to nutritional supplements or exercise. Loss of skeletal muscle mass and force increases the risk of falls, impairs mobility, and leads to reduced quality of life. In a previous study, we demonstrated that taking in Alaska pollock protein (APP) for only 7 d increased the gastrocnemius muscle mass in rats. This study was conducted to identify hypertrophic myofibers and analyze how hypertrophy occurs within them. Twenty male rats were randomly divided into two groups and administered a diet of casein or APP for 7 d. The expression of each myosin heavy chain (MyHC) isoform in a cross-sectional area was then measured. MyHC IIb and IIx isoforms exhibited hypertrophic features in the gastrocnemius muscles of the APP-fed rats. Furthermore, comprehensive proteomic analyses were conducted to identify changes in protein expression due to muscle hypertrophy. Our results, evaluated by pathway analyses, indicated that the activity of the growth factor signaling pathway was significantly impacted by APP consumption. Moreover, APP could promote protein synthesis by activating the protein kinase B/mechanistic target of the rapamycin signaling pathway, which is also promoted by exercise.

Dietary Alaska Pollack Protein Induces Acute and Sustainable Skeletal Muscle Hypertrophy in Rats.

Our previous studies suggested that Alaska pollack protein (APP) intake increases skeletal muscle mass and that it may cause a slow-to-fast shift in muscle fiber type in rats fed a high-fat diet after 56 days of feeding. In this study, we explored whether dietary APP induces acute and sustainable skeletal muscle hypertrophy in rats fed a normal-fat diet. Male 5-week-old Sprague-Dawley rats were divided into four groups and fed a purified ingredient-based high-fat diet or a purified ingredient-based normal-fat diet with casein or APP, containing the same amount of crude protein. Dietary APP significantly increased gastrocnemius muscle mass (105~110%) after 2, 7 days of feeding, regardless of dietary fat content. Rats were separated into two groups and fed a normal-fat diet with casein or APP. Dietary APP significantly increased gastrocnemius muscle mass (110%) after 56 days of feeding. Dietary APP significantly increased the cross-sectional area of the gastrocnemius skeletal muscle and collagen-rich connective tissue after 7 days of feeding. It decreased the gene expression of Mstn /Myostatin, Trim63/MuRF1, and Fbxo32/atrogin-1, but not other gene expression, such as serum IGF-1 after 7 days of feeding. No differences were observed between casein and APP groups with respect to the percentage of Type I, Type IIA, and Type IIX or IIB fibers, as determined by myosin ATPase staining after 7 days of feeding. In the similar experiment, the puromycin-labeled peptides were not different between dietary casein and APP after 2 days of feeding. These results demonstrate that APP induces acute and sustainable skeletal muscle hypertrophy in rats, regardless of dietary fat content. Dietary APP, as a daily protein source, may be an approach for maintaining or increasing muscle mass.

Fish Protein Ingestion Induces Neural, but Not Muscular Adaptations, Following Resistance Training in Young Adults.

Purpose: Nutritional supplementation in conjunction with exercise is of interest for the prevention or improvement of declines in motor performances in older adults. An understanding of the effects on both young and older adults contributes to its effective application. We investigated the effect of fish protein ingestion with resistance training on neural and muscular adaptations in young adults using interventions and assessments that have already been tested in older adults. Methods: Eighteen young adults underwent 8 weeks of isometric knee extension training. During the intervention, nine participants ingested 5 g of fish protein (n = 9, Alaska pollack protein, APP), and the other nine participants ingested casein as a control (n = 9, CAS) in addition to daily meals. Before, during, and after the intervention, the isometric knee extension force, lower extremity muscle mass, and motor unit firing pattern of knee extensor muscles were measured. Results: Maximum voluntary contraction (MVC) was significantly increased in both APP and CAS groups from 0 weeks to 4, 6, and 8 weeks of intervention (p < 0.001), but there were no significant differences between the groups (p = 0.546-0.931). Muscle mass was not significantly changed during the intervention in either group (p = 0.250-0.698). Significant changes in motor unit firing rates (p = 0.02 and 0.029 for motor units recruited at 20-40% of MVC and at 40-60%) were observed following the intervention in the APP but not CAS (p = 0.120-0.751) group. Conclusions: These results suggest that dietary fish protein ingestion changes motor unit adaptations following resistance training in young adults.

In vitro and in vivo evaluation of the efficacy of bovine colostrum against human rotavirus infection.

We found that skimmed and concentrated bovine late colostrum (SCBLC) obtained from normal cows at 6-7 d after parturition exhibited high potency in inhibiting replication of human rotavirus (HRV) in vitro. Furthermore, prophylactic oral administration of SCBLC once before inoculation of HRV prevented the development of diarrhea in suckling mice in vivo. SCBLC from normal cows might be useful in the prevention of HRV-induced severe gastroenteritis in immunocompromised hosts.

Binding of Norovirus virus-like particles (VLPs) to human intestinal Caco-2 cells and the suppressive effect of pasteurized bovine colostrum on this VLP binding.

Noroviruses (NoVs), which cannot be grown in cell culture, are a major infectious agent of gastroenteritis. An in vitro assay system was established for the evaluation of NoV binding to enterocytes using virus-like particles (VLPs) produced in a baculovirus system expressing a NoV VP1 capsid protein. After confirmation of the purity by MS analysis, VLPs were incubated with human intestinal Caco-2 cells. NoV VLPs were detected clearly by confocal laser microscopy only on a certain population of Caco-2 cells, and were semi-quantified by immunoblotting of cell lysates. Then the suppressive effect of pasteurized bovine colostrum was analyzed on the VLP binding to Caco-2 cells by immunoblotting. The colostrum reduced VLP binding in a dose-dependent manner, at about 50% suppression with 12.5 microg of the colostral proteins. Furthermore, the colostrum contained IgG antibodies reacting to VLPs, suggesting that cross-reactive antibodies in the bovine colostrums block human NoV binding to intestinal cells.

Modulation of Neural and Muscular Adaptation Processes During Resistance Training by Fish Protein Ingestions in Older Adults.

Assessments of both neural and muscular adaptations during interventions would provide valuable information for developing countermeasures to age-related muscle dysfunctions. We investigated the effect of fish protein ingestion on training-induced neural and muscular adaptations in older adults. Twenty older adults participated 8 weeks of isometric knee extension training intervention. The participants were divided into two groups who took fish protein (n = 10, Alaska pollack protein, APP) or casein (n = 10, CAS). Maximal muscle strength during knee extension, lower extremity muscle mass (body impedance method), and motor unit firing pattern of knee extensor muscle (high-density surface electromyography) were measured before, during, and after the intervention. Muscle strength were significantly increased in both CAS (124.7 ± 5.8%) and APP (117.1 ± 4.4%) after intervention (p < .05), but no significant differences between the groups were observed (p > .05). Significant increases in lower extremity muscle mass from 0 to 8 weeks were demonstrated only for APP (102.0 ± 3.2, p < .05). Greater changes in motor unit firing pattern following intervention were represented in CAS more than in APP. These results suggest that nutritional supplementations could modulate neural and muscular adaptations following resistance training and fish protein ingestion preferentially induces muscular adaptation without the detectable neural adaptation in older adults.

Hypothalamic-pituitary-adrenal axis differentially responses to morning and evening psychological stress in healthy subjects.

AIM: The hypothalamic-pituitary-adrenal (HPA) axis responds to changing environmental demands including psychological stressors. The aim of the present study was to assess whether the time of day effects on the acute response of HPA axis activity to acute psychological stress. METHOD: We studied 27 healthy young subjects. The subjects were participated two experiments as follows. In the first experiment, subjects were instructed to keep their regular sleep schedule for 2 weeks which were measured by using a wrist-worn activity monitor. Afterward, to evaluate a diurnal rhythm of salivary cortisol, eight saliva samples were collected during waking period every 2 hours from when the subjects woke up. In the second experiment, the subjects were randomly assigned to two groups. The Trier Social Stress Test (TSST) was performed either in the morning (n = 14) or in the evening (n = 13). We measured diurnal rhythm of salivary cortisol and stress response of salivary cortisol and heart rate by the TSST. Morning and evening tests were started at 2 hours and 10 hours after woke up, respectively. RESULTS: All subjects showed a normal diurnal rhythm of salivary cortisol concentration, with a peak in the morning immediately after awaking and a minimum in the evening. The salivary cortisol response after the TSST was significantly increased from the prestress level in the morning but not in the evening. CONCLUSION: The HPA response to acute psychological stress was more pronounced in the morning than in the evening, correlating with the circadian regulation of cortisol synthesis.