Development of a PCL-PEO double network colorimetric pH sensor using electrospun fibers containing Hibiscus rosa sinensis extract and silver nanoparticles for food monitoring.

Major anthocyanin, cyanidin-3-sophoroside (318.1 mg/mL), and other minor copigments were identified in the ethanol extract of Hibiscus rosa sinensis. The extracts can be coelectrospun with polycaprolactone and polyethylene oxide into fiber mats and were sensitive to pH changes from 1 to 13 with a unique color code (ΔE > 5). The pH sensor was used to monitor shrimp quality under isothermal conditions to obtain the respective activation energy (Ea in kJ/mol) of the sensors' color-change response (20.2), measured pH (20.6), and trimethylamine nitrogen (24.6), indole (27.1), and total microbial counts (30.8). Together with the Pearson correlation coefficient, the results showed high correlations between the sensors' color change and other quality parameters (p < 0.001). The regression equation developed by conducting the kinetic analysis was also suitable for predicting shrimp quality at refrigeration temperatures (4-10 °C) and can be used as a marker to monitor shrimp quality by visually inspecting the item condition.

Identification and characterization of immunomodulatory peptides from pepsin-soy protein hydrolysates.

To obtain immunomodulatory peptides from isolated soy protein (ISP), pepsin was selected to prepare hydrolysates and 4-h treatment (Pepsin-ISPH4h) showed the highest yield and immunomodulatory activities. The Pepsin-ISPH4h was sequentially fractionated by 30, 10 and 1-kDa molecular weight cut-off (MWCO) membranes, in which 1-kDa MWCO permeate (1P) exhibited the most significant enhancement of phagocytosis activity without causing excessive inflammation as compared with Pepsin-ISPH4h. To further purify and enhance the immunomodulatory activity, 1P was distinct by high-performance liquid chromatography equipped with a reverse-phase column and in vivo immunomodulatory activity of fractions was examined in mice. Fraction 1 (F1) significantly elevated phagocytosis activity of mice spleen macrophages and neutrophils. However, increase of phagocytosis activity did not result from the induction of macrophages M1 or M2 polarization. The immunomodulatory peptide sequence, EKPQQQSSRRGS, from F1 was identified by LC-MS/MS. Phagocytosis activity and macrophage M1 polarization were elevated by synthetic peptide treatment. Hence, our results indicated that isolated soy protein hydrolysates prepared by pepsin could provide a source of peptides with immunomodulatory effects.

Effects of Dietary Inclusion of Dry Hydrastis canadensis on Laying Performance, Egg Quality, Serum Biochemical Parameters and Cecal Microbiota in Laying Hens.

Alternative growth promoters are able to not only effectively replace the traditional use of antibiotics but also provide additional health benefits for livestock and reduce food safety concerns. This study investigated the effects of dry Hydrastis canadensis on the laying performance and fecal microbial community of laying hens. Twenty-four Lohmann (LSL, white layer strain) hens were reared from 40 to 48 weeks of age and randomly allotted to four dietary treatments (six birds/treatment). The dietary treatments comprised a basal diet with no treatment as control, a basal diet plus 0.6% powder of dry Hydrastis canadensis roots (R) or leaves (L), and a basal diet plus 0.6% powder of a mixture of dry Hydrastis canadensis roots and leaves (1:1, LR). No mortality was observed in the whole experimental period. The results indicated that albumen height in the LR group was significantly greater than that in the control group. The diet supplemented with Hydrastis canadensis had no significant effects on egg production rate, egg weight, eggshell strength, eggshell thickness, Haugh unit, or yolk height during the whole experimental phase. However, principal coordinate analysis, comparative heat map analysis, and cluster dendrogram analysis of cecal microbiota showed distinct clusters among the groups treated with Hydrastis canadensis and the control group. Regarding blood biochemical parameters, serum cholesterol levels were significantly lower in all Hydrastis canadensis-treated groups compared with those in the control group. Moreover, serum low-density lipoprotein levels were lower in hens supplemented with the leaf of Hydrastis canadensis. The abundances of the phyla Fusobacteria and Kiritimatiellaeota were increased (p < 0.05) in laying hens fed with 0.6% Hydrastis canadensis leaves, whereas the abundance of the phylum Firmicutes in cecum digesta decreased in response to treatment with Hydrastis canadensis roots and leaves. The relative abundance of the Fusobacterium genus was higher in the LR group compared with that in the control. On the contrary, we found a different trend in the Synergistes genus. The potential influences of these microbiota on the performance of laying hens were discussed. The results demonstrate that Hydrastis canadensis can improve the egg albumen height and modulate the cecum digesta microbiota composition of laying hens.

Evaluation of the Efficacy of Supplementation with Planox® Lemon Verbena Extract in Improving Oxidative Stress and Muscle Damage: A Randomized Double-Blind Controlled Trial.

Excessive exercise load can cause muscle soreness and fatigue, as well as inflammation and oxidative stress. Lemon verbena (Aloysia triphylla; Lippia citriodora) is often used as a spice in tea or beverages. Its leaves are rich in polyphenols, which have antioxidant and anti-inflammatory bioactivities. In the present study, we investigated whether supplementation with Planox® lemon verbena extract (LVE) could improve muscle damage and biochemical indicators after exhaustive exercise challenge. All subjects (30 males and 30 females) underwent a double-blind trial and were randomly divided into a placebo group (0 mg/human/day) and an LVE supplement group (400 mg/human/day), with gender-equal distribution. All subjects started supplementation 10 days before exhaustive exercise and continued it until all tests were completed. Before the intervention, after the exhaustive exercise, and on the following 3 days, the participants underwent 12-minute Cooper running/walking; blood collection; assessments of pain, muscle stiffness, maximum jump heights, and isometric maximum muscle strength. The results showed that supplementation with LVE effectively increased GPx and reduced CK, IL-6, 8-OHdG and muscle pain after the exhaustive exercise, but it had significant effect on strength recovery. In summary, LVE is a safe and edible natural plant extract that can reduce muscle damage and soreness after exercise. This trial was registered at clinicaltrials.gov as NCT04742244.

Shikonin Inhibits Der p 2-Induced Cytokine and Chemokine Expression in Dendritic Cells in Patients with Atopic Dermatitis.

Atopic dermatitis (AD) is a common inflammatory skin disorder. Shikonin, the active component of Lithospermum erythrorhizon extract, exhibits anti-inflammatory effects. The objective of the present study was to investigate the effect of shikonin on proinflammatory cytokines and chemokine in patients with AD. Ten patients with AD who were allergic to house dust mite (HDM) and seven healthy controls were recruited in this study. Peripheral blood mononuclear cells were isolated, and CD14+ cells were further selected and differentiated to dendritic cells. Dendritic cells stimulated using Der p 2, the major HDM allergen, were cotreated with shikonin for 24 hours, and dexamethasone was used as a control. Culture supernatants were collected, and proinflammatory cytokine and chemokine concentrations were analyzed using a multiplex assay system. Shikonin significantly inhibited Der p 2-induced expression of interleukin (IL)-6, IL-9, and IL-17A; monocyte chemoattractant protein (MCP)-1; macrophage inflammatory protein (MIP)-1α; MIP-1ß; and Chemokine (C-C motif) ligand 5 (RANTES). The inhibitory effects of shikonin on IL-9, MIP-1ß, and RANTES expression were stronger than those of dexamethasone. Therefore, Shikonin can be considered a promising drug for AD treatment because it inhibits different inflammatory cytokines expression.

Protein hydrolysate from potato confers hepatic-protection in hamsters against high fat diet induced apoptosis and fibrosis by suppressing Caspase-3 and MMP2/9 and by enhancing Akt-survival pathway.

BACKGROUND: A potato protein hydrolysate, APPH is a potential anti-obesity diet ingredient. Since, obesity leads to deterioration of liver function and associated liver diseases, in this study the effect of APPH on high fat diet (HFD) associated liver damages was investigated. METHODS: Six week old male hamsters were randomly separated to six groups (n = 8) as control, HFD (HFD fed obese), L-APPH (HFD + 15 mg/kg/day of APPH), M-APPH (HFD + 30 mg/kg/day), H-APPH (HFD + 75 mg/kg/day of APPH) and PB (HFD + 500 mg/kg/day of probucol). HFD fed hamsters were administered with APPH 50 days through oral gavage. The animals were euthanized and the number of apoptotic nuclei in liver tissue was determined by TUNEL staining and the extent of interstitial fibrosis was determined by Masson's trichrome staining. Modulation in the molecular events associated with apoptosis and fibrosis were elucidated from the western blotting analysis of the total protein extracts. RESULTS: Hamsters fed with high fat diet showed symptoms of liver damage as measured from serum markers like alanine aminotransferase and aspartate aminotransferase levels. However a 50 day long supplementation of APPH effectively ameliorated the effects of HFD. HFD also modulated the expression of survival and apoptosis proteins in the hamster liver. Further the HFD groups showed elevated levels of fibrosis markers in liver. The increase in fibrosis and apoptosis was correlated with the increase in the levels of phosphorylated extracellular signal-regulated kinases (pERK1/2) revealing a potential role of ERK in the HFD mediated liver damage. However APPH treatment reduced the effect of HFD on the apoptosis and fibrosis markers considerably and provided hepato-protection. CONCLUSION: APPH can therefore be considered as an efficient therapeutic agent to ameliorate high fat diet related liver damages.

Oral administration of alcalase potato protein hydrolysate-APPH attenuates high fat diet-induced cardiac complications via TGF-ß/GSN axis in aging rats.

Consumption of high fat diet (HFD) is associated with increased cardiovascular risk factors among elderly people. Aging and obesity induced-cardiac remodeling includes hypertrophy and fibrosis. Gelsolin (GSN) induces cardiac hypertrophy and TGF-ß, a key cytokine, which induces fibrosis. The relationship between TGF-ß and GSN in aging induced cardiac remodeling is still unknown. We evaluated the expressions of TGF-ß and GSN in HFD fed 22 months old aging SD rats, followed by the administration of either probucol or alcalase potato protein hydrolysate (APPH). Western blotting and Masson trichrome staining showed that APPH (45 and 75 mg/kg/day) and probucol (500 mg/kg/day) treatments significantly reduced the aging and HFD-induced hypertrophy and fibrosis. Echocardiograph showed that the performance of the hearts was improved in APPH, and probucol treated HFD aging rats. Serum from all rats was collected and H9c2 cells were cultured with collected serums separately. The GSN dependent hypertrophy was inhibited with an exogenous TGF-ß in H9c2 cells cultured in HFD+ APPH treated serum. Thus, we propose that along with its role in cardiac fibrosis, TGF-ß also acts as an upstream activator of GSN dependent hypertrophy. Hence, TGF-ß in serum could be a promising therapeutic target for cardiac remodeling in aging and/or obese subjects.

Lipolysis-Stimulating Peptide from Soybean Protects Against High Fat Diet-Induced Apoptosis in Skeletal Muscles.

Obesity is generally associated with low-grade chronic inflammation that involves the recruitment of macrophages and other inflammation factors to the adipocytes of obese individuals. Tumor necrosis factor-alpha (TNF-α), a cytokine associated with systemic inflammation, is elevated in conditions of obesity. TNF-α is an important factor that plays an important role in skeletal muscle wasting. Apoptosis of myonuclei contributes to the loss of muscle mass and therefore plays an important role in skeletal muscle atrophy. In mouse models that were fed a high fat diet (HFD), a lipolysis-stimulating peptide-VHVV (purified from hydrolysate resulting from flavourzyme treatment of soy protein) was found to reduce HFD-related apoptotic effects in mice skeletal muscle and potentially control atrophy. HFD fed mice had heavier body weight than those fed with normal chow, and VHVV administration restricted lipid accumulation in muscle tissues of mice fed with HFD but increased nutrient uptake. Moreover, specific concentrations of VHVV regulated TNF-α expression that was elevated by HFD, suppressed apoptosis-related proteins and regulated the proteins of lipid metabolism.

Lithospermum erythrorhizon extract inhibits Der p2-induced inflammatory response through alleviation of thymic stromal lymphopoietin, nuclear factor Kappa B, and inflammasome expression in human bronchial epithelial cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Lithospermum erythrorhizon (LE) and Angelica sinensis (AS), widely used in several folk medicine for wound, pus discharge and dermatitis for the history of several hundred years in Asian countries. AIM OF STUDY: To investigate the therapeutic effect of LE and AS on Der p2-induced inflammatory response in human bronchial epithelial (BEAS-2B) cells. METHODS: The effects of Der p2 stimulation on thymic stromal lymphopoietin (TSLP), the nuclear factor kappa B (NF-κB) pathway, the inflammasome (specifically, the apoptosis speck-like protein [ASC] and nod-like receptor 3 [NLRP3]), Caspase-1 and the signal transducer and activator of transcription (STAT) 3 pathway were evaluated in the human bronchial epithelial (BEAS-2B) cells. RESULTS: The results indicated that LE, AS, and LE+AS reduced TSLP, I kappa B kinase-α, and NLRP3 levels; LE and AS reduced Caspase-1; LE and LE+AS also reduced NF-κB p50, NF-κB p65, ASC, and STAT3 levels. CONCLUSION: Both LE and AS aqueous extracts exert anti-inflammatory effects in Der p2-stimulated BEAS-2B cells. These effects may involve multiple mechanisms, including the inhibition of TSLP production as well as the suppression of IKKα, Caspase-1 and NLRP3; however, additional studies are warranted to elucidate the underlying mechanisms.

Do demographic characteristics influence the eating competence of elderly Taiwanese?

BACKGROUND AND OBJECTIVES: To investigate the association between demographic characteristics and eating competence (EC), "the behaviour and attitudinal conceptualization of eating", among elderly Taiwanese. METHODS AND STUDY DESIGN: Random questionnaire survey. Overall, 564 questionnaires were analyzed, and independent t-tests, analysis of variance, chi-square tests, were used to compare the differences in the EC scores when stratified by various demographic variables. RESULTS: The mean score of both EC and non-EC (NEC) for all respondents was 29.3. Younger respondents (65-70 years old) reported significantly higher EC scores than did older respondents (71-76 years old) (p<0.05). Similarly, respondents with high school or a higher education level reported higher scores than did respondents with elementary or a lower education level (p<0.001), and respondents with partners had higher scores than did those with no partners (p<0.001). Respondents who lived in urban areas reported higher EC scores than those living in rural areas (p<0.001). No significant gender differences were found in both EC and NEC scores. CONCLUSIONS AND IMPLICATIONS: Thirty-seven percent of elderly Taiwanese were not eating competent. Older respondents with higher education levels, those with partners, and those residing in urban areas showed significantly higher EC scores. Nutritional practitioners should therefore develop nutrition education programs tailored for those with low EC scores.

Lipolysis stimulating peptides of potato protein hydrolysate effectively suppresses high-fat-diet-induced hepatocyte apoptosis and fibrosis in aging rats.

BACKGROUND: Non-alcoholic fatty liver disease (NAFLD) is one of the most common outcomes of obesity and is characterized by the accumulation of triglycerides, increased tissue apoptosis, and fibrosis. NAFLD is more common among elderly than in younger age groups, and it causes serious hepatic complications. OBJECTIVE: In this study, alcalase treatment derived potato protein hydrolysate (APPH) with lipolysis-stimulating property has been evaluated for its efficiency to provide hepato-protection in a high-fat-diet (HFD)-fed aging rats. DESIGN: Twenty-four-month-old SD rats were randomly divided into six groups (n=8): aged rats fed with standard chow, HFD-induced aged obese rats, HFD with low-dose (15 mg/kg/day) APPH treatment, HFD with moderate (45 mg/kg/day) APPH treatment, HFD with high (75 mg/kg/day) APPH treatment, and HFD with probucol. RESULTS: APPH was found to reduce the NAFLD-related effects in rat livers induced by HFD and all of the HFD-fed rats exhibited heavier body weight than those with control chow diet. However, the HFD-induced hepatic fat accumulation was effectively attenuated in rats administered with low (15 mg/kg/day), moderate (45 mg/kg/day), and high (75 mg/kg/day) doses of APPH. APPH oral administration also suppressed the hepatic apoptosis- and fibrosis-related proteins induced by HFD. CONCLUSIONS: Our results thus indicate that APPH potentially attenuates hepatic lipid accumulation and anti-apoptosis and fibrosis effects in HFD-induced rats. APPH may have therapeutic potential in the amelioration of NAFLD liver damage.

The Heart Protection Effect of Alcalase Potato Protein Hydrolysate Is through IGF1R-PI3K-Akt Compensatory Reactivation in Aging Rats on High Fat Diets.

The prevalence of obesity is high in older adults. Alcalase potato protein hydrolysate (APPH), a nutraceutical food, might have greater benefits and be more economical than hypolipidemic drugs. In this study, serum lipid profiles and heart protective effects were evaluated in high fat diet (HFD) induced hyperlipidemia in aging rats treated with APPH (15, 45 and 75 mg/kg/day) and probucol (500 mg/kg/day). APPH treatments reduced serum triacylglycerol (TG), total cholesterol (TC), and low density lipoprotein (LDL) levels to the normal levels expressed in the control group. Additionally, the IGF1R-PI3K-Akt survival pathway was reactivated, and Fas-FADD (Fas-associated death domain) induced apoptosis was inhibited by APPH treatments (15 and 45 mg/kg/day) in HFD aging rat hearts. APPH (75 mg/kg/day) rather than probucol (500 mg/kg/day) treatment could reduce serum lipids without affecting HDL expression. The heart protective effect of APPH in aging rats with hyperlipidemia was through lowering serum lipids and enhancing the activation of the compensatory IGF1R-PI3K-Akt survival pathway.

Hepatoprotective Effects of Swimming Exercise against D-Galactose-Induced Senescence Rat Model.

This study investigates whether a 12-week swimming exercise training can prevent liver damage or senescence associated biomarkers in an experimental aging model in rats. Twenty-three male Sprague-Dawley rats were divided into four groups: vehicle treatment with sedentary control (C, n = 6), aging induction with sedentary (A, n = 6), vehicle treatment with swimming exercise (SW, n = 5), and aging induction with swimming exercise (A + SW, n = 6). Rats in groups A and AS received intraperitoneal d-galactose injections (150 mg/kg/day) for 12 weeks to induce aging. Rats in groups SW and A + SW were subjected to swimming exercise training for 12 weeks. Body weight, liver weight, epididymal fat mass, blood biochemistry, and liver pathology were performed at the end of the experiment. Hepatic senescence protein markers such as ß-galactosidase, p53, and p21, as well as the inflammatory mediator, IL-6, were examined. The d-galactose-treated rats exhibited increases in AST and γ -GT plasma levels and ß-galactosidase protein expression compared to the control group. Swimming exercise significantly reduced BW, epididymal fat mass, γ -GT activity, and p53, p21, and IL-6 protein levels compared to the aging group. These results suggest that a 12-week swimming exercise program suppresses senescence markers and downregulates inflammatory mediator in the liver tissues of d-galactose-induced aging rats.

Purification and identification of lipolysis-stimulating peptides derived from enzymatic hydrolysis of soy protein.

The aim of this study was to purify and identify lipolysis-stimulating peptides derived from Flavourzyme®-soy protein isolate (SPI) hydrolysate (F-SPIH). Glycerol release was employed as a marker for lipolysis in 3T3-L1 adipocytes. A higher glycerol release represents a better lipolysis-stimulating activity. The peptide fraction with highest glycerol release obtained from F-SPIH fractionated by sequential ultrafiltration membranes was further purified using gel filtration chromatography and two steps of reverse-phase high-performance liquid chromatography. The peptides were identified using liquid chromatography-tandem mass spectrometry (LC/MS/MS). Three lipolysis-stimulating peptides were obtained, and the amino acid sequences were ILL, LLL and VHVV, respectively. The in vitro effect of gastrointestinal proteases on lipolysis-stimulating activity of synthetic ILL, LLL and VHVV, respectively, was also investigated. The result suggested that the gastrointestinal protease did not affect lipolysis-stimulating activity of the three novel peptides, which reveals their potential to act as anti-obesity ingredients.

Influence of alanine uptake on Staphylococcus aureus surface charge and its susceptibility to two cationic antibacterial agents, nisin and low molecular weight chitosan.

Low molecular weight chitosan (LMWC) and nisin, recognized as cationic antibacterial agents (CAAs), inhibit bacterial growth by interacting with the anionically charged cell wall. In this study, alanine uptake significantly reduced the anionic cell surface charge, as determined by the zeta potential (ZP) measurements, of Staphylococcus aureus, resulting from the incorporation of d-alanine into the cell wall. Minimum inhibitory concentration (MIC) tests and growth inhibition curves revealed that LMWC and nisin possessed inverse antibacterial activity against three strains of S. aureus, depending on the strains' net charge. A twofold reduction in the MIC value of nisin was obtained against S. aureus, inoculated in a 1.0% d- or l-alanine-augmented trypticase soy broth medium. A flocculation test demonstrated that neutralizing the anionic surface charge using d-alanine reduced the adsorption of S. aureus onto LMWC. Furthermore, the reduced surface net charge could enhance the colonization capacity of S. aureus on glass.

Solvent selection and optimization of α-chymotrypsin-catalyzed synthesis of N-Ac-Phe-Tyr-NH2 using mixture design and response surface methodology.

A peptide, N-Ac-Phe-Tyr-NH(2) , with angiotensin I-converting enzyme (ACE) inhibitor activity was synthesized by an α-chymotrypsin-catalyzed condensation reaction of N-acetyl phenylalanine ethyl ester (N-Ac-Phe-OEt) and tyrosinamide (Tyr-NH(2) ). Three kinds of solvents: a Tris-HCl buffer (80 mM, pH 9.0), dimethylsulfoxide (DMSO), and acetonitrile were employed in this study. The optimum reaction solvent component was determined by simplex centroid mixture design. The synthesis efficiency was enhanced in an organic-aqueous solvent (Tris-HCl buffer: DMSO: acetonitrile = 2:1:1) in which 73.55% of the yield of N-Ac-Phe-Tyr-NH(2) could be achieved. Furthermore, the effect of reaction parameters on the yield was evaluated by response surface methodology (RSM) using a central composite rotatable design (CCRD). Based on a ridge max analysis, the optimum condition for this peptide synthesis included a reaction time of 7.4 min, a reaction temperature of 28.1°C, an enzyme activity of 98.9 U, and a substrate molar ratio (Phe:Tyr) of 1:2.8. The predicted and the actual (experimental) yields were 87.6 and 85.5%, respectively. The experimental design and RSM performed well in the optimization of synthesis of N-Ac-Phe-Tyr-NH(2) , so it is expected to be an effective method for obtaining a good yield of enzymatic peptide. © 2012 American Institute of Chemical Engineers Biotechnol. Prog., 2012.

Enhancing the lipolysis-stimulating activity of soy protein using limited hydrolysis with Flavourzyme and ultrafiltration.

The aim of this study was to explore the lipolysis-stimulating activity of soy protein isolate (SPI) hydrolysate using 3T3-L1 adipocytes. Intracellular triglyceride residue (TR) was employed as a marker for lipolysis in cells. The lower TR represents the better lipolysis-stimulating activity. SPI was hydrolysed with Flavourzyme for 2 h to obtain the hydrolysate FH2h, which showed lipolysis-stimulating activity in adipocytes at 400-1600 ppm levels. The sequential fractionation of FH2h with 30-0.3 kDa molecular weight cut-off (MWCO) membranes in order to obtain a 1 kDa retentate resulted in further enhancement of its lipolysis-stimulating activity in the cells. The TR decreased significantly from 2.73 to 2.30 µmole/mg protein at the 400 ppm level (p<0.05). Based on the western immunoblot and immunostaining analysis, the 1 kDa retentate promotes lipolysis by increasing phosphorylation and translocation of the hormone-sensitive lipase in 3T3-L1 adipocytes.