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1.
J Agric Food Chem ; 71(4): 2070-2081, 2023 Feb 01.
Article in English | MEDLINE | ID: mdl-36652684

ABSTRACT

It is well known that repeated exposure to phenolic compounds (PCs) raises astringency perception. However, the link between this increase and the oral cavity's interactions with salivary proteins (SPs) and other oral constituents is unknown. To delve deeper into this connection, a flavonoid-rich green tea extract was tested in a series of exposures to two oral cell-based models using a tongue cell line (HSC3) and a buccal mucosa cell line (TR146). Serial exposures show cumulative PC binding to all oral models at all concentrations of the green tea extract; however, the contribution for the first and second exposures varies. The tongue mucosal pellicle (HSC3-Mu-SP) may contribute more to first-stage astringency (retaining 0.15 ± 0.01 mg mL-1 PCs at the first exposure), whereas the buccal mucosal pellicle (TR146-Mu-SP) retained significantly less (0.08 ± 0.02 mg mL-1). Additionally, increased salivary volume (SV+), which simulates the stimulation of salivary flow brought by a food stimulus, significantly enhances PC binding, particularly for TR146 cells: TR46-Mu-SP_SV+ bound significantly higher total PC concentration (0.17 ± 0.02 mg mL-1) than the model without increased salivary volume TR146-Mu-SP_SV- (0.09 ± 0.03 mg mL-1). This could be associated with a higher contribution of these oral cells for astringency perception during repeated exposures. Furthermore, PCs adsorbed in the first exposure to cell monolayer models (+TR146 and +HSC3) change the profile of PCs bound to these models in the second exposure. Regarding the structure binding activity, PCs with a total higher number of hydroxyl groups were more bound by the models containing SP. Regarding the SP, basic proline-rich proteins (bPRPs) may be involved in the increased perception of astringency upon repeated exposures. The extent of bPRP precipitation by PCs in mucosal pellicle models for both cell lines (HSC3 and TR146) in the second exposure (76 ± 13 and 83 ± 6%, respectively) was significantly higher than in the first one (25 ± 14 and 5 ± 6%, respectively).


Subject(s)
Astringents , Flavonoids , Aspergillus fumigatus/metabolism , Astringents/chemistry , Azoles , Drug Resistance, Fungal , Flavonoids/metabolism , Fungal Proteins/metabolism , Phenols/metabolism , Saliva/chemistry , Salivary Proteins and Peptides/metabolism , Tea/metabolism , Mouth
2.
Sci Rep ; 12(1): 10246, 2022 06 17.
Article in English | MEDLINE | ID: mdl-35715581

ABSTRACT

Horticultural therapy (HT) has been reported to be beneficial to mental and physical health. This study investigated the effects of HT on the psychological status and mucosal immunity of elderly individuals. Twenty-four participants aged 70-93 were recruited from residential facilities and adult day-care services. Six different HT activities were designed and guided by licensed instructors who performed saliva collection and helped the participants complete the questionnaires before and after each activity. The sleep quality scores were collected during the 6 weeks of HT activities. Saliva was collected and analyzed to determine the concentrations of immunoglobulin A (IgA), lactoferrin, chromogranin A (CgA), α-amylase (AA) and total protein (TP). Comparisons of the questionnaire scores between preactivity and postactivity showed that feelings of satisfaction and happiness were significantly enhanced after each activity. In addition, sleep quality was significantly improved after the 6-week course of HT activities. Regarding mucosal immunity, the preactivity IgA and IgA/TP were significantly increased at week 3 and week 6; in addition, the ratio of lactoferrin/TP was significantly decreased at week 6 compared to week 1. The postactivity AA and CgA levels were significantly enhanced at weeks 2, 3 and 5 compared to the corresponding preactivity levels. In conclusions, HT activities significantly improved the happiness, satisfaction, well-being and sleep quality of the elderly. Moreover, mucosal immunity proteins, including IgA, lactoferrin, CgA and AA, were significantly increased.


Subject(s)
Horticultural Therapy , Adult , Aged , Amylases/metabolism , Biomarkers/metabolism , Chromogranin A/metabolism , Humans , Immunity, Mucosal , Immunoglobulin A/metabolism , Lactoferrin/metabolism , Saliva/metabolism , Salivary Proteins and Peptides/metabolism , Sleep Quality , alpha-Amylases/metabolism
3.
Angew Chem Int Ed Engl ; 60(10): 5348-5356, 2021 03 01.
Article in English | MEDLINE | ID: mdl-33345438

ABSTRACT

Blood feeding arthropods, such as leeches, ticks, flies and mosquitoes, provide a privileged source of peptidic anticoagulant molecules. These primarily operate through inhibition of the central coagulation protease thrombin by binding to the active site and either exosite I or exosite II. Herein, we describe the rational design of a novel class of trivalent thrombin inhibitors that simultaneously block both exosites as well as the active site. These engineered hybrids were synthesized using tandem diselenide-selenoester ligation (DSL) and native chemical ligation (NCL) reactions in one-pot. The most potent trivalent inhibitors possessed femtomolar inhibition constants against α-thrombin and were selective over related coagulation proteases. A lead hybrid inhibitor possessed potent anticoagulant activity, blockade of both thrombin generation and platelet aggregation in vitro and efficacy in a murine thrombosis model at 1 mg kg-1 . The rational engineering approach described here lays the foundation for the development of potent and selective inhibitors for a range of other enzymatic targets that possess multiple sites for the disruption of protein-protein interactions, in addition to an active site.


Subject(s)
Anticoagulants/therapeutic use , Platelet Aggregation Inhibitors/therapeutic use , Salivary Proteins and Peptides/therapeutic use , Thrombosis/drug therapy , Amblyomma/chemistry , Animals , Anopheles/chemistry , Anticoagulants/chemical synthesis , Anticoagulants/metabolism , Catalytic Domain , Humans , Male , Mice, Inbred C57BL , Platelet Aggregation Inhibitors/chemical synthesis , Platelet Aggregation Inhibitors/metabolism , Protein Binding , Protein Engineering , Salivary Proteins and Peptides/chemical synthesis , Salivary Proteins and Peptides/metabolism , Thrombin/chemistry , Thrombin/metabolism , Tsetse Flies/chemistry
4.
Sci Rep ; 10(1): 12638, 2020 07 28.
Article in English | MEDLINE | ID: mdl-32724226

ABSTRACT

Phenolic compounds (PC) are linked to astringency sensation. Astringency studies typically use simple models, with pure PC and/or proteins, far from what is likely to occur in the oral cavity. Different oral models have been developed here, comprising different oral epithelia (buccal mucosa (TR146) and tongue (HSC-3)) and other main oral constituents (human saliva and mucosal pellicle). These models, were used to study the interaction with two PC extracts, one rich in flavanols (a green tea extract) and one rich in anthocyanins (a red wine extract). It was observed that within a family of PC, the PC seem to have a similar binding to both TR146 and HSC-3 cell lines. When the oral constituents occur altogether, flavanols showed a higher interaction, driven by the salivary proteins. Conversely, anthocyanins showed a lower interaction when the oral constituents occur altogether, having a higher interaction only with oral cells. Epigallocatechin gallate, epicatechin gallate, epigallocatechin-3-O(3-O-methyl) gallate were the flavanols with the highest interaction. For the studied anthocyanins (delphinidin-3-glucoside, peonidin-3-glucoside, petunidin-3-glucoside and malvidin-3-glucoside), there was not a marked difference on their interaction ability. Overall, the results support that the different oral constituents can have a different function at different phases of food (PC) intake. These differences can be related to the perception of different astringency sub-qualities.


Subject(s)
Anthocyanins/metabolism , Flavonols/metabolism , Mouth Mucosa/metabolism , Plant Extracts/metabolism , Salivary Proteins and Peptides/metabolism , Tea/chemistry , Tongue/metabolism , Wine/analysis , Adult , Cells, Cultured , Female , Humans , In Vitro Techniques , Male , Saliva/metabolism , Young Adult
5.
BMC Oral Health ; 20(1): 169, 2020 06 12.
Article in English | MEDLINE | ID: mdl-32532263

ABSTRACT

BACKGROUND: Recent preventive strategies for dental caries focus on targeting the mechanisms underlying biofilm formation, including the inhibition of bacterial adhesion. A promising approach to prevent bacterial adhesion is to modify the composition of acquired salivary pellicle. This in vitro study investigated the effect and possible underlying mechanism of pellicle modification by casein phosphopeptide (CPP) on Streptococcus mutans (S. mutans) initial adhesion, and the impact of fluoride on the efficacy of CPP. METHODS: The salivary pellicle-coated hydroxyapatite (s-HA) discs were treated with phosphate buffered saline (negative control), heat-inactivated 2.5% CPP (heat-inactivated CPP), 2.5% CPP (CPP) or 2.5% CPP supplemented with 900 ppm fluoride (CPP + F). After cultivation of S. mutans for 30 min and 2 h, the adherent bacteria were visualized by scanning electron microscopy (SEM) and quantitatively evaluated using the plate count method. Confocal laser scanning microscopy (CLSM) was used to evaluate the proportions of total and dead S. mutans. The concentrations of total, free, and bound calcium and fluoride in the CPP and fluoride-doped CPP solutions were determined. The water contact angle and zeta potential of s-HA with and without modification were measured. The data were statistically analyzed using one-way ANOVA followed by a Turkey post hoc multiple comparison test. RESULTS: Compared to the negative control group, the amount of adherent S. mutans significantly reduced in the CPP and CPP + F groups, and was lowest in the CPP + F group. CLSM analysis showed that there was no statistically significant difference in the proportion of dead S. mutans between the four groups. Water contact angle and zeta potential of s-HA surface significantly decreased in the CPP and CPP + F groups as compared to the negative control group, and both were lowest in the CPP + F group. CONCLUSIONS: Pellicle modification by CPP inhibited S. mutans initial adhesion to s-HA, possibly by reducing hydrophobicity and negative charge of the s-HA surface, and incorporating fluoride into CPP further enhanced the anti-adhesion effect.


Subject(s)
Bacterial Adhesion/drug effects , Caseins/pharmacology , Dental Caries/prevention & control , Durapatite/chemistry , Fluorides/pharmacology , Phosphopeptides/pharmacology , Saliva/chemistry , Streptococcus mutans/drug effects , Biofilms , Coated Materials, Biocompatible/chemistry , Dental Caries Susceptibility , Humans , Saliva/microbiology , Salivary Proteins and Peptides/metabolism , Streptococcus mutans/isolation & purification , Streptococcus mutans/physiology , Turkey
6.
BMC Genomics ; 21(1): 331, 2020 Apr 29.
Article in English | MEDLINE | ID: mdl-32349672

ABSTRACT

BACKGROUND: Salivary cell secretion (SCS) plays a critical role in blood feeding by medicinal leeches, making them of use for certain medical purposes even today. RESULTS: We annotated the Hirudo medicinalis genome and performed RNA-seq on salivary cells isolated from three closely related leech species, H. medicinalis, Hirudo orientalis, and Hirudo verbana. Differential expression analysis verified by proteomics identified salivary cell-specific gene expression, many of which encode previously unknown salivary components. However, the genes encoding known anticoagulants have been found to be expressed not only in salivary cells. The function-related analysis of the unique salivary cell genes enabled an update of the concept of interactions between salivary proteins and components of haemostasis. CONCLUSIONS: Here we report a genome draft of Hirudo medicinalis and describe identification of novel salivary proteins and new homologs of genes encoding known anticoagulants in transcriptomes of three medicinal leech species. Our data provide new insights in genetics of blood-feeding lifestyle in leeches.


Subject(s)
Genome , Hirudo medicinalis/genetics , Salivary Proteins and Peptides/genetics , Animals , Anticoagulants/metabolism , Gene Expression Profiling , Gene Expression Regulation , Hirudo medicinalis/metabolism , Leeches/classification , Leeches/genetics , Leeches/metabolism , Proteomics , Saliva/metabolism , Salivary Proteins and Peptides/metabolism
7.
Carbohydr Polym ; 236: 116044, 2020 May 15.
Article in English | MEDLINE | ID: mdl-32172858

ABSTRACT

In this study, water and chelator-soluble pectic polysaccharide fractions were obtained from white grape skins, aiming to study their impact on the interaction between low polymerized grape seed procyanidins and salivary proteins. Water and chelator-soluble polysaccharide fractions were composed by uronic acids and neutral sugars, mainly arabinose and galactose, with water polysaccharide fraction showing a higher amount of branched pectic polysaccharides. Both polysaccharide fractions were able to mitigate salivary protein-procyanidin interactions, by a competition mechanism, resulting in a decrease of the amount of precipitated protein. Water polysaccharide fraction was the most effective in inhibiting salivary protein precipitation, especially for acidic proline-rich proteins, due to the higher affinity to interact with procyanidins (KA = 22222 M-1 and KA = 365 M-1 for water and chelator polysaccharides, respectively). The interaction between polysaccharides and procyanidins showed to be mainly governed by hydrophobic effect.


Subject(s)
Biflavonoids/metabolism , Catechin/metabolism , Pectins/chemistry , Proanthocyanidins/metabolism , Salivary Proteins and Peptides/metabolism , Vitis/chemistry , Biflavonoids/analysis , Biflavonoids/isolation & purification , Catechin/analysis , Catechin/isolation & purification , Fruit/chemistry , Humans , Pectins/analysis , Pectins/isolation & purification , Proanthocyanidins/analysis , Proanthocyanidins/isolation & purification , Protein Binding/drug effects
8.
J Breath Res ; 13(2): 026004, 2019 03 01.
Article in English | MEDLINE | ID: mdl-30530941

ABSTRACT

Mucoadhesive tablets containing herbal formulation have been previously shown to reduce oral malodour. The aim of the present in vitro study was to test the effect of the mucoadhesive agent hydroxyethylcellulose (HEC) added to a liquid phase herbal extract formulation on the mucoadhesive retention of the active ingredients and their effect against malodour production. Experimental oral biofilms were grown on mucin coated glass slides treated with liquid phase solutions of herbal extract with or without HEC as well as saline and 0.2% chlorhexidine as controls. Biofilms were quantified for volatile sulfide compounds (VSC) producing bacteria using CLSM and sampled for a salivary incubation assay to test for malodour production (odour judge), VSC production (Halimeter) and salivary protein degradation (SDS-PAGE). Results showed that the addition of HEC to the herbal extracts solution has significantly increased its mucin retained activity against malodour producing bacteria and their resulting malodour and VSC production and proteolytic activities. Taken together, results of the present study suggest that the addition of HEC to a liquid phase herbal extract solution may increase its bioavailability time and efficacy. However, due to the limitations of this in vitro study additional clinical investigations are needed.


Subject(s)
Adhesives/pharmacology , Biofilms/drug effects , Cellulose/analogs & derivatives , Mouth/microbiology , Mucus/chemistry , Plant Extracts/pharmacology , Sulfides/analysis , Animals , Bacteria/drug effects , Bacteria/metabolism , Cellulose/pharmacology , Humans , Salivary Proteins and Peptides/metabolism , Sus scrofa , Volatilization
9.
Food Funct ; 9(9): 4948-4958, 2018 Sep 19.
Article in English | MEDLINE | ID: mdl-30182113

ABSTRACT

Cancer patients receiving chemotherapy often experience taste and smell abnormalities (TSA). To date, the underlying molecular mechanisms of this frequent side-effect have not been determined and effective treatments are not available. This study assessed the feasibility of lactoferrin (LF) supplementation as a treatment for TSA and investigate the related mechanisms through salivary proteome analysis. Nineteen cancer patients with established TSA following chemotherapy administration were enrolled in this study. Cancer patients and additional 12 healthy subjects took LF supplements, 3 tablets per day (250 mg per tablet), for 30 days. Saliva was collected at three timepoints: baseline, 30-day LF supplementation, and 30-day post-LF supplementation. Patient's TSA level, salivary proteome, and salivary minerals at each LF treatment stage were analyzed. High TSA level was associated with high concentration of salivary Fe and loss of critical salivary immune proteins. LF supplementation significantly decreased the concentration of salivary Fe (P = 0.025), increased the abundance (P < 0.05) of salivary α-amylase and Zn-α-2-GP, and led to an overall increase of expression (≥2-fold changes) of immune proteins including immunoglobulin heavy chain, annexin A1, and proteinase inhibitor. Abundance of α-amylase and SPLUNC2 were further increased (P < 0.05) at 30-day post-LF supplementation in cancer patients. At the same time, total TSA score was significantly reduced (P < 0.001) in chemotherapy patients. This study demonstrated the feasibility of developing lactoferrin supplementation as a treatment to reduce TSA caused by chemotherapy and improve cancer patient's oral immunity.


Subject(s)
Antineoplastic Agents/adverse effects , Dietary Supplements , Lactoferrin/therapeutic use , Olfaction Disorders/therapy , Saliva/metabolism , Salivary Proteins and Peptides/metabolism , Taste Disorders/therapy , Aged , Antioxidants/adverse effects , Antioxidants/therapeutic use , Biomarkers/metabolism , Dietary Supplements/adverse effects , Feasibility Studies , Female , Humans , Immunity, Mucosal/drug effects , Immunoglobulin Heavy Chains/metabolism , Iron/metabolism , Lactoferrin/adverse effects , Male , Middle Aged , Minerals/metabolism , Olfaction Disorders/chemically induced , Olfaction Disorders/metabolism , Olfaction Disorders/physiopathology , Oxidative Stress/drug effects , Proteomics/methods , Saliva/enzymology , Saliva/immunology , Salivary Elimination/drug effects , Self Report , Severity of Illness Index , Taste Disorders/chemically induced , Taste Disorders/metabolism , Taste Disorders/physiopathology
10.
PLoS One ; 12(11): e0187069, 2017.
Article in English | MEDLINE | ID: mdl-29095857

ABSTRACT

Hyposalivation reduces the patient quality of life, as saliva is important for maintaining oral health. Current treatments for hyposalivation are limited to medications such as the muscarinic receptor agonists, pilocarpine and cevimeline. However, these therapies only provide temporary relief. Therefore, alternative therapies are essential to restore salivary gland function. An option is to use bioengineered scaffolds to promote functional salivary gland regeneration. Previous studies demonstrated that the laminin-111 protein is critical for intact salivary gland cell cluster formation and organization. However, laminin-111 protein as a whole is not suitable for clinical applications as some protein domains may contribute to unwanted side effects such as degradation, tumorigenesis and immune responses. Conversely, the use of synthetic laminin-111 peptides makes it possible to minimize the immune reactivity or pathogen transfer. In addition, it is relatively simple and inexpensive as compared to animal-derived proteins. Therefore, the goal of this study was to demonstrate whether a 20 day treatment with laminin-111-derived peptide conjugated fibrin hydrogel promotes tissue regeneration in submandibular glands of a wound healing mouse model. In this study, laminin-111-derived peptide conjugated fibrin hydrogel significantly accelerated formation of salivary gland tissue. The regenerated gland tissues displayed not only structural but also functional restoration.


Subject(s)
Fibrin/chemistry , Hydrogels , Laminin/pharmacology , Peptides/pharmacology , Salivary Glands/drug effects , Animals , Female , Materials Testing , Mice , Mice, Inbred C57BL , Saliva/metabolism , Salivary Glands/physiology , Salivary Proteins and Peptides/metabolism
11.
J Agric Food Chem ; 65(10): 2147-2154, 2017 Mar 15.
Article in English | MEDLINE | ID: mdl-28225606

ABSTRACT

The interaction of astringent substances with salivary proteins, which results in protein precipitation, is considered a key event in the molecular mechanism underlying the oral sensation of puckering astringency. As the chemical nature of orally active astringents is diverse and the knowledge of their interactions with salivary proteins rather fragmentary, human whole saliva samples were incubated with suprathreshold and isointensity solutions of the astringent polyphenol (-)-epigallocatechin gallate, the multivalent metal salt iron(III) sulfate, the amino-functionalized polysaccharide chitosan, and the basic protein lysozyme. After separation of the precipitated proteins, the proteins affected by the astringents were identified and relatively quantified for the first time by complementary bottom-up and top-down mass spectrometry-based proteomics approaches. Major salivary target proteins, which may be involved in astringency perception, are reported here for each astringent stimulus.


Subject(s)
Astringents/metabolism , Mouth/metabolism , Salivary Proteins and Peptides/metabolism , Adult , Astringents/chemistry , Electrophoresis, Polyacrylamide Gel , Female , Humans , Male , Mass Spectrometry , Proteomics , Salivary Proteins and Peptides/chemistry , Taste , Young Adult
12.
Acupunct Med ; 33(6): 451-6, 2015 Dec.
Article in English | MEDLINE | ID: mdl-26449884

ABSTRACT

BACKGROUND: A previous study has reported that low-frequency (LF) electroacupuncture (EA) influences salivary secretory immunoglobulin A (sIgA) and the autonomic nervous system (ANS). The ANS is known to control the secretion volume of sIgA; however, the effect of high-frequency (HF) EA on salivary sIgA has not been determined. We investigated whether HF EA affects salivary sIgA levels and the ANS. METHOD: Sixteen healthy subjects were randomly classified into two groups: a control group and an EA group. After a 5 min rest, subjects in the EA group received EA at 100 Hz bilaterally at LI4 and LI11 for 15 min before resting for a further 40 min post-stimulation. Subjects in the control group rested for a total of 60 min. Measurements of the ANS and sIgA levels in both groups were made before, immediately after, 20 min after, and 40 min after rest or 15 min EA treatment. HF and LF components of heart rate variability were analysed as markers of ANS function. LF/HF ratio and HF were taken as indices of sympathetic and parasympathetic nerve activity, respectively. Salivary protein concentrations and sIgA levels were determined by Bradford protein assay and ELISA, respectively. RESULTS: LF/HF ratio was significantly increased immediately after EA. HF was significantly increased at 20 min after EA and sIgA level was significantly increased at 40 min after EA. In addition, HF and salivary sIgA level were positively correlated with each another. CONCLUSIONS: HF EA exerted sequential positive effects on sympathetic nerve activity, parasympathetic nerve activity, and salivary sIgA level (immediately and after 20 and 40 min, respectively). HF EA may increase salivary sIgA levels by influencing parasympathetic nerve activity.


Subject(s)
Autonomic Nervous System , Electroacupuncture , Immunoglobulin A, Secretory/metabolism , Acupuncture Points , Adult , Aged , Aged, 80 and over , Heart Rate , Humans , Middle Aged , Saliva/immunology , Salivary Proteins and Peptides/metabolism , Young Adult
13.
PLoS One ; 9(9): e108372, 2014.
Article in English | MEDLINE | ID: mdl-25264771

ABSTRACT

The salivary mucins that include MUC5B (gel-forming) and MUC7 (non-gel-forming) are major contributors to the protective mucus barrier in the oral cavity, and it is possible that dietary components may influence barrier properties. We show how one dietary compound, the green tea polyphenol epigallocatechin gallate (EGCG), can substantially alter the properties of both the polymeric MUC5B network and monomeric MUC7. Using rate-zonal centrifugation, MUC5B in human whole saliva and MUC5B purified from saliva sedimented faster in the presence of EGCG. The faster sedimentation by EGCG was shown to be greater with increasing MUC5B concentration. Particle tracking microrheology was employed to determine the viscosity of purified MUC5B solutions and showed that for MUC5B solutions of 200-1600 µg/mL, EGCG caused a significant increase in mucin viscosity, which was greater at higher MUC5B concentrations. Visualisation of the changes to the MUC5B network by EGCG was performed using atomic force microscopy, which demonstrated increased aggregation of MUC5B in a heterogeneous manner by EGCG. Using trypsin-resistant, high-molecular weight oligosaccharide-rich regions of MUC5B and recombinant N-terminal and C-terminal MUC5B proteins, we showed that EGCG causes aggregation at the protein domains of MUC5B, but not at the oligosaccharide-rich regions of the mucin. We also demonstrated that EGCG caused the majority of MUC7 in human whole saliva to aggregate. Furthermore, purified MUC7 also underwent a large increase in sedimentation rate in the presence of EGCG. In contrast, the green tea polyphenol epicatechin caused no change in the sedimentation rate of either MUC5B or MUC7 in human whole saliva. These findings have demonstrated how the properties of the mucin barrier can be influenced by dietary components. In the case of EGCG, these interactions may alter the function of MUC5B as a lubricant, contributing to the astringency (dry puckering sensation) of green tea.


Subject(s)
Catechin/analogs & derivatives , Catechin/metabolism , Mucin-5B/metabolism , Mucins/metabolism , Saliva/metabolism , Salivary Proteins and Peptides/metabolism , Camellia sinensis , Centrifugation, Zonal , Diet , Humans , Microscopy, Atomic Force , Polyphenols/metabolism , Protein Aggregates , Protein Structure, Tertiary , Tea
14.
PLoS One ; 9(1): e87580, 2014.
Article in English | MEDLINE | ID: mdl-24498143

ABSTRACT

The aim of this study was to investigate the short-term effects of green tea consumption on selected salivary defense proteins, antibacterial capacity and anti-oxidation activity in taekwondo (TKD) athletes, following intensive training. Twenty-two TKD athletes performed a 2-hr TKD training session. After training, participants ingested green tea (T, caffeine 6 mg/kg and catechins 22 mg/kg) or an equal volume of water (W). Saliva samples were collected at three time points: before training (BT-T; BT-W), immediately after training (AT-T; AT-W), and 30 min after drinking green tea or water (Rec-T; Rec-W). Salivary total protein, immunoglobulin A (SIgA), lactoferrin, α-amylase activity, free radical scavenger activity (FRSA) and antibacterial capacity were measured. Salivary total protein, lactoferrin, SIgA concentrations and α-amylase activity increased significantly immediately after intensive TKD training. After tea drinking and 30 min rest, α-amylase activity and the ratio of α-amylase to total protein were significantly higher than before and after training. In addition, salivary antibacterial capacity was not affected by intense training, but green tea consumption after training enhanced salivary antibacterial capacity. Additionally, we observed that salivary FRSA was markedly suppressed immediately after training and quickly returned to pre-exercise values, regardless of which fluid was consumed. Our results show that green tea consumption significantly enhances the activity of α-amylase and salivary antibacterial capacity.


Subject(s)
Anti-Bacterial Agents/metabolism , Exercise/physiology , Martial Arts/physiology , Saliva/metabolism , Salivary Proteins and Peptides/metabolism , Tea/metabolism , Adult , Antioxidants/metabolism , Athletes , Drinking/physiology , Female , Free Radical Scavengers/metabolism , Humans , Immunoglobulin A/metabolism , Lactoferrin/metabolism , Male , Young Adult , alpha-Amylases/metabolism
15.
Anal Biochem ; 451: 28-30, 2014 Apr 15.
Article in English | MEDLINE | ID: mdl-24463016

ABSTRACT

The heterologous recombinant expression of proteins in Escherichia coli without start-methionine is a common problem. The nitrophorin 7 heme properties and function strongly depend on the accurate N-terminal amino acid sequence. Leading protein expression into the periplasm by fusion with the leader peptide pelB yields functional protein; however, the folded protein sticks to the cell debris. Therefore, the periplasmic fraction was dissolved in guanidinium chloride and folded by a drop-in method. Separation from impurities including residual pelB-nitrophorin 7 required establishing an unconventional chromatographic technique using calcium-loaded Chelating Sepharose as cation exchanger and elution by a linear CaCl2 gradient.


Subject(s)
Escherichia coli/metabolism , Hemeproteins/metabolism , Rhodnius/metabolism , Salivary Proteins and Peptides/metabolism , Animals , Chromatography, Ion Exchange , Codon, Initiator , Guanidine/chemistry , Hemeproteins/chemistry , Hemeproteins/genetics , Methionine/metabolism , Periplasm/metabolism , Polysaccharide-Lyases/genetics , Polysaccharide-Lyases/metabolism , Protein Folding , Recombinant Fusion Proteins/biosynthesis , Recombinant Fusion Proteins/chemistry , Recombinant Fusion Proteins/isolation & purification , Salivary Proteins and Peptides/chemistry , Salivary Proteins and Peptides/genetics , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
16.
Article in English | MEDLINE | ID: mdl-24111288

ABSTRACT

Such popular head care procedures as shampooing and scalp massages provide physical and mental relaxation. However, they place a big burden such as chapped hands on beauticians and other practitioners. Based on our robot hand technology, we have been developing a head care robot. In this paper, we quantitatively evaluated its relaxation effect using the following biosignals: accelerated plethymography (SDNN, HF/TP, LF/HF), heart rate (HR), blood pressure, salivary amylase (sAA) and peripheral skin temperature (PST). We compared the relaxation of our developed head care robot with the head care provided by nurses. In our experimental result with 54 subjects, the activity of the autonomic nerve system changed before and after head care procedures performed by both a human nurse and our proposed robot. Especially, in the proposed robot, we confirmed significant differences with the procedure performed by our proposed head care robot in five indexes: HF/TP, LF/HF, HR, sAA, and PST. The activity of the sympathetic nerve system decreased, because the values of its indexes significantly decreased: LF/HF, HR, and sAA. On the other hand, the activity of the parasympathetic nerve system increased, because of the increase of its indexes value: HF/TP and PST. Our developed head care robot provided satisfactory relaxation in just five minutes of use.


Subject(s)
Head , Massage , Robotics , Adult , Amylases/metabolism , Blood Pressure/physiology , Female , Heart Rate/physiology , Humans , Male , Massage/instrumentation , Massage/methods , Plethysmography/methods , Robotics/instrumentation , Robotics/methods , Salivary Proteins and Peptides/metabolism , Skin Temperature/physiology
17.
J Food Sci ; 77(5): H96-H104, 2012 May.
Article in English | MEDLINE | ID: mdl-22497429

ABSTRACT

We tested the hypothesis that rats adapt to the iron absorption inhibitory effects of tea by modifying the expression of salivary proteins. Thirty-six weanling rats were allocated into 6 groups. Two control groups were fed a semipurified diet containing 20 mg Fe(2+)/kg diet. Two groups were fed spray dried green tea infusion mixed into the diet (28.6 g tea/kg diet) and 2 groups were fed the control diet with a twice daily gavage of a tea solution (0.25 g tea/mL). Saliva samples were collected in 3 groups (control, gavage, and oral) on day 8 (acute) and in the remaining groups on day 31 (chronic). Iron absorption was assessed using a (58)Fe(3+) tracer administered on day 1 (acute) and day 24 (chronic). 2D gel electrophoresis and mass spectrometry were used to assess the composition of the saliva proteome. There was no significant difference in iron absorption between the 3 groups on either day 1 or day 24. Salivary proline-rich proteins and submandibular gland secretory protein increased to a greater extent in the oral group than in the gavage group, when compared to control, within the same exposure time period. Amylase, chitinase, deoxyribonuclease, cysteine-rich secretory protein 1, and parotid secretory protein all decreased to a greater extent in the oral tea group, compared to the control, within the same exposure time period. Our results show that green tea did not decrease iron absorption in rats but it did have a marked effect on the saliva proteome when given orally.


Subject(s)
Iron/pharmacokinetics , Proteome/chemistry , Saliva/chemistry , Tea/chemistry , Absorption , Amylases/genetics , Amylases/metabolism , Animal Feed , Animals , Chitinases/genetics , Chitinases/metabolism , Deoxyribonucleases/genetics , Deoxyribonucleases/metabolism , Diet , Eating , Liver/drug effects , Liver/metabolism , Male , Membrane Glycoproteins/genetics , Membrane Glycoproteins/metabolism , Proteome/analysis , Proteomics/methods , Rats , Rats, Sprague-Dawley , Salivary Proline-Rich Proteins , Salivary Proteins and Peptides/genetics , Salivary Proteins and Peptides/metabolism , Trypsin/metabolism
18.
J Agric Food Chem ; 60(15): 3966-72, 2012 Apr 18.
Article in English | MEDLINE | ID: mdl-22440016

ABSTRACT

Condensed tannins are a group of polyphenols that are associated with the astringency sensation, as they readily interact and precipitate salivary proteins. As this interaction is affected by carbohydrates, the aim of this work was to study the effect of some carbohydrates used in the food industry [arabic gum (AG), pectin, and poligalacturonic acid (PGA)] on the salivary proteins/grape seed procyanidins interaction. This was assessed monitoring the salivary proteins that remain soluble in the presence of condensed tannins with the addition of carbohydrates (HPLC) and analysis of the respective precipitates (SDS-PAGE). The results show that pectin was the most efficient in inhibiting protein/tannin precipitation, followed by AG and PGA. The results suggest that pectin and PGA exert their effect by formation of a ternary complex protein/polyphenol/carbohydrate, while AG competes with proteins for tannin binding (competition mechanism). The results also point out that both hydrophilic and hydrophobic interactions are important for the carbohydrate effects.


Subject(s)
Gum Arabic/chemistry , Pectins/chemistry , Proanthocyanidins/chemistry , Salivary Proteins and Peptides/chemistry , Down-Regulation , Gum Arabic/metabolism , Humans , Pectins/metabolism , Proanthocyanidins/metabolism , Protein Binding , Salivary Proteins and Peptides/metabolism
19.
Eur J Oral Sci ; 120(2): 132-9, 2012 Apr.
Article in English | MEDLINE | ID: mdl-22409219

ABSTRACT

Green tea is a popular drink throughout the world, and it contains various components, including the green tea polyphenol (-)-epigallocatechin gallate (EGCG). Tea interacts with saliva upon entering the mouth, so the interaction between saliva and EGCG interested us, especially with respect to EGCG-protein binding. SDS-PAGE revealed that several salivary proteins were precipitated after adding EGCG to saliva. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) peptide mass fingerprinting indicated that the major proteins precipitated by EGCG were alpha-amylase, S100, and cystatins. Surface plasmon resonance revealed that EGCG bound to alpha-amylase at dissociation constant (K(d)) = 2.74 × 10(-6) M, suggesting that EGCG interacts with salivary proteins with a relatively strong affinity. In addition, EGCG inhibited the activity of alpha-amylase by non-competitive inhibition, indicating that EGCG is effective at inhibiting the formation of fermentable carbohydrates involved in caries formation. Interestingly, alpha-amylase reduced the antimicrobial activity of EGCG against the periodontal bacterium Aggregatibacter actinomycetemcomitans. Therefore, we considered that EGCG-salivary protein interactions might have both protective and detrimental effects with respect to oral health.


Subject(s)
Catechin/analogs & derivatives , Dental Caries/prevention & control , Salivary Proteins and Peptides/metabolism , Tea , alpha-Amylases/analysis , Adult , Aggregatibacter actinomycetemcomitans/drug effects , Anti-Bacterial Agents/antagonists & inhibitors , Catechin/metabolism , Catechin/pharmacology , Cystatins/antagonists & inhibitors , Dietary Carbohydrates/antagonists & inhibitors , Female , Humans , Male , Middle Aged , Protein Binding , Proteome/analysis , Saliva/enzymology , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Tea/chemistry , Young Adult , alpha-Amylases/antagonists & inhibitors
20.
Biomaterials ; 32(35): 9168-79, 2011 Dec.
Article in English | MEDLINE | ID: mdl-21906807

ABSTRACT

In the oral cavity, omnipresent salivary protein films (pellicle) mediate bacterial adhesion and biofilm formation on natural tissues as well as on artificial implant surfaces, which may cause serious infectious diseases like periimplantitis. The purpose of this in vitro study was to investigate the adsorption/desorption behaviour of human saliva on model surfaces grafted with polyamidoamine (PAMAM) dendrimer molecules compared to self-assembled monolayers (SAMs) exhibiting the same terminal functions (-NH(2), -COOH) by two complementary analytical methods. Furthermore, the role of saliva conditioning of PAMAM and analogous SAM modifications on the adhesion of Streptococcus gordonii DL1, an early oral colonizer, was investigated. In contrast to SAMs, PAMAM-grafted surfaces showed reduced streptococcal adherence in the absence of pre-adsorbed saliva similar to the level obtained for poly(ethylene glycol) (PEG) coatings. Moreover, coatings of PAMAM-NH(2) maintained their bacteria-repellent behaviour even after saliva-conditioning. As a general outcome, it was found that lower amounts of protein adsorbed on PAMAM coatings than on analogous SAMs. Since this study demonstrates that covalently bound PAMAM dendrimers can modulate the oral bacterial response, this approach has significant potential for the development of anti-adhesive biomaterial surfaces that are conditioned with proteinaceous films.


Subject(s)
Bacterial Adhesion/drug effects , Dendrimers/chemistry , Dendrimers/pharmacology , Models, Chemical , Salivary Proteins and Peptides/metabolism , Silicon/pharmacology , Streptococcus gordonii/physiology , Adsorption/drug effects , Humans , Hydrogen-Ion Concentration/drug effects , Isoelectric Point , Microscopy, Atomic Force , Quartz Crystal Microbalance Techniques , Static Electricity , Streptococcus gordonii/cytology , Streptococcus gordonii/drug effects , Surface Properties/drug effects , Water/chemistry
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