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1.
Food Res Int ; 178: 113964, 2024 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-38309881

RESUMEN

Astringency corresponds to the sensation of dryness and roughness that is experienced in the oral cavity in association with the interaction between salivary proteins and food polyphenols. In this study, the phenolic composition of seven varietal wines, the intensity of astringency they evoke and the physicochemical reactivity of these wines with whole human saliva were evaluated. Phenolic composition of wines was characterized by spectrophotometry and HPLC chromatography. Intensity of astringency was evaluated by trained sensory panels. Saliva from a single volunteer subject was used to assess wine-saliva interactions. To this end, binary mixtures were produced at different v/v wine/saliva ratios and each of them assayed for the ability of the salivary protein to diffuse on a cellulose membrane (diffusion test) and to remain in solution (precipitation test). Physicochemical reactivities between wine components and the protein fraction of saliva were contrasted against the astringency and the phenolic profile of each varietal wine. The study supports the view that astringency depends on physicochemical interactions between two complex matrices -wine and saliva- and not between some of their particular components.


Asunto(s)
Vino , Humanos , Vino/análisis , Saliva/química , Gusto , Astringentes/análisis , Polifenoles/análisis , Fenoles/análisis , Proteínas y Péptidos Salivales/análisis
2.
Microbiome ; 11(1): 69, 2023 03 31.
Artículo en Inglés | MEDLINE | ID: mdl-37004076

RESUMEN

BACKGROUND: Dental erosion is a disease of the oral cavity where acids cause a loss of tooth enamel and is defined as having no bacterial involvement. The tooth surface is protected from acid attack by salivary proteins that make up the acquired enamel pellicle (AEP). Bacteria have been shown to readily degrade salivary proteins, and some of which are present in the AEP. This study aimed to explore the role of bacteria in dental erosion using a multi-omics approach by comparing saliva collected from participants with dental erosion and healthy controls. RESULTS: Salivary proteomics was assessed by liquid-chromatography mass spectrometry (LC-MS) and demonstrated two altered AEP proteins in erosion, prolactin inducible protein (PIP), and zinc-alpha-2 glycoprotein (ZAG). Immunoblotting further suggested that degradation of PIP and ZAG is associated with erosion. Salivary microbiome analysis was performed by sequencing the bacterial 16S rRNA gene (V1-V2 region, Illumina) and showed that participants with dental erosion had a significantly (p < 0.05) less diverse microbiome than healthy controls (observed and Shannon diversity). Sequencing of bacterial mRNA for gene expression (Illumina sequencing) demonstrated that genes over-expressed in saliva from erosion participants included H + proton transporter genes, and three protease genes (msrAB, vanY, and ppdC). Salivary metabolomics was assessed using nuclear magnetic resonance spectrometry (NMR). Metabolite concentrations correlated with gene expression, demonstrating that the dental erosion group had strong correlations between metabolites associated with protein degradation and amino acid fermentation. CONCLUSIONS: We conclude that microbial proteolysis of salivary proteins found in the protective acquired enamel pellicle strongly correlates with dental erosion, and we propose four novel microbial genes implicated in this process. Video Abstract.


Asunto(s)
Erosión de los Dientes , Humanos , Erosión de los Dientes/metabolismo , Proteolisis , Disbiosis/metabolismo , ARN Ribosómico 16S/metabolismo , Saliva , Proteínas y Péptidos Salivales/análisis , Proteínas y Péptidos Salivales/metabolismo , Péptido Hidrolasas
3.
J Exp Bot ; 74(21): 6874-6888, 2023 11 21.
Artículo en Inglés | MEDLINE | ID: mdl-37103882

RESUMEN

The brown planthopper (BPH) Nilaparvata lugens (Stål) is a main pest on rice. It secretes saliva to regulate plant defense responses, when penetrating rice plant and sucking phloem sap through its stylet. However, the molecular mechanisms of BPH salivary proteins regulating plant defense responses remain poorly understood. A N. lugens DNAJ protein (NlDNAJB9) gene was highly expressed in salivary glands, and the knock down of NlDNAJB9 significantly enhanced honeydew excretion and fecundity of the BPH. NlDNAJB9 could induce plant cell death, and the overexpression of NlDNAJB9 gene in Nicotiana benthamiana induced calcium signaling, mitogen-activated protein kinase (MAPK) cascades, reactive oxygen species (ROS) accumulation, jasmonic acid (JA) hormone signaling and callose deposition. The results from different NlDNAJB9 deletion mutants indicated that the nuclear localization of NlDNAJB9 was not necessary to induce cell death. The DNAJ domain was the key region to induce cell death, and the overexpression of DNAJ domain in N. benthamiana significantly inhibited insect feeding and pathogenic infection. NlDNAJB9 might interact indirectly with NlHSC70-3 to regulate plant defense responses. NlDNAJB9 and its orthologs were highly conserved in three planthopper species, and could induce ROS burst and cell death in plants. Our study provides new insights into the molecular mechanisms of insect-plant interactions.


Asunto(s)
Hemípteros , Oryza , Animales , Especies Reactivas de Oxígeno/metabolismo , Saliva/química , Hemípteros/fisiología , Inmunidad de la Planta/genética , Proteínas y Péptidos Salivales/análisis , Proteínas y Péptidos Salivales/metabolismo , Oryza/genética
4.
Molecules ; 28(6)2023 Mar 09.
Artículo en Inglés | MEDLINE | ID: mdl-36985492

RESUMEN

Beer is one of the most consumed beverages worldwide with unique organoleptic properties. Bitterness and astringency are well-known key features and, when perceived with high intensity, could lead to beer rejection. Most studies on beer astringency and bitterness use sensory assays and fail to study the molecular events that occur inside the oral cavity responsible for those perceptions. This work focused on deepening this knowledge based on the interaction of salivary proteins (SP) and beer phenolic compounds (PCs) and their effect toward these two sensory attributes. The astringency and bitterness of four different beers were assessed by a sensory panel and were coupled to the study of the SP changes and PC profile characterization of beers. The human SP content was measured before (basal) and after each beer intake using HPLC analysis. The beers' PC content and profile were determined using Folin-Ciocalteu and LC-MS spectrometry, respectively. The results revealed a positive correlation between PCs and astringency and bitterness and a negative correlation between SP changes and these taste modalities. Overall, the results revealed that beers with higher PC content (AAL and IPA) are more astringent and bitter than beers with a lower PC content (HL and SBO). The correlation results suggested that an increase in whole SP content, under stimulation, should decrease astringency and bitterness perception. No correlation was found between the changes in specific families of SP and astringency and bitterness perception.


Asunto(s)
Astringentes , Gusto , Humanos , Astringentes/análisis , Cerveza/análisis , Percepción del Gusto , Fenoles/análisis , Proteínas y Péptidos Salivales/análisis
5.
Clin Oral Investig ; 27(1): 285-298, 2023 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-36149517

RESUMEN

OBJECTIVES: This study aimed to characterise diurnal dynamics of salivary peptidome and variations induced by sampling procedures. MATERIALS AND METHODS: A supervised short-term longitudinal study was conducted amongst ten healthy participants. Saliva samples were collected by different procedures (stimulated/unstimulated conditions, forepart/midstream segments) on three consecutive days. The peptidome compositions of saliva samples were analysed using matrix-assisted laser desorption/ionisation-time of flight-mass spectrometry (MALDI-TOF MS). RESULTS: The salivary peptidome exhibited a stable trend generally, even though some diurnal dynamics happened in aspects of both overall structure and certain single peptides. The results indicated saliva samples collected under unstimulated and stimulated conditions have significantly different structures of peptidome, whilst the peptidome profile of stimulated saliva was more abundant than that of unstimulated saliva. It was also indicated that the midstream segment effect might exist in the segmented process of saliva sampling. CONCLUSIONS: In summary, salivary peptidome was able to maintain stability though some dynamic changes might happen within a short-term period. Stimulated and unstimulated saliva samples had significantly different peptidome profiles, whilst the stimulated whole saliva was a larger pool of low molecular weight peptides. CLINICAL RELEVANCE: The stability of the salivary peptidome highlights the reliability of salivary peptidome as a source of diagnostic biomarker. We recommend keeping one collection condition (stimulated/unstimulated) consistently within one study on salivary peptidome. Stimulated whole saliva would be preferred if more abundant peptidome profile is needed.


Asunto(s)
Péptidos , Saliva , Humanos , Estudios Longitudinales , Reproducibilidad de los Resultados , Péptidos/análisis , Péptidos/química , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/métodos , Saliva/química , Proteínas y Péptidos Salivales/análisis
6.
J Dent ; 129: 104389, 2023 02.
Artículo en Inglés | MEDLINE | ID: mdl-36526084

RESUMEN

OBJECTIVES: To evaluate and compare the proteomic profile of acquired pellicle on smooth bovine tooth and tooth-coloured restorative materials, including resin composite (RC), glass ionomer cement (GIC), and casein phosphopeptide-amorphous calcium phosphate modified GIC (CPP-ACP GIC). METHODS: Two-hour in situ pellicles on tooth/materials specimens mounted in oral appliances worn by ten healthy adults were investigated. Pellicle proteins and corresponding unstimulated whole saliva were quantitatively analysed through liquid chromatography-tandem mass spectrometry. RESULTS: Significantly higher amounts of protein were adsorbed onto tooth surface than restorative materials tested (4.11 ± 0.69 vs. 2.54 ± 0.38/2.98 ± 0.80/3.01 ± 0.37 µg, RC/GIC/CPP-ACP GIC). From the ten participants, 1,444 (487-1,086/person), 1,454 (645-1,051/person), 1,731 (454-1,475/person), or 1,597 (423-1,261/person) pellicle proteins were detected at least once on bovine tooth, RC, GIC, or CPP-ACP GIC, respectively, and with 1,072 (304-793/person) salivary proteins identified. Comparative quantification revealed minor differences between tooth and restorative materials pellicle profiles. High inter-individual variations in pellicle protein composition were demonstrated. Compared to the salivary protein profile, 214/57 proteins showed significantly increased/decreased abundance in pellicle formed on at least one substrate (fold change > 3.325/fold change < 0.301). Gene Ontology enrichment analysis showed some pellicle proteins detected with increased affinity to tooth/material surface were identified as being related to "calcium-dependent protein binding" or "cell-cell adhesion mediator activity". CONCLUSION: Similar protein quantity and composition was observed in 2 h in situ pellicles formed on different smooth restorative material surfaces. The proteomic profile of pellicles appeared distinct from that of the corresponding unstimulated whole saliva. CLINICAL SIGNIFICANCE: Host backgrounds appeared more influential on the proteomic profile of the in situ acquired pellicle than the underlying substrate characteristics among systemically and orally healthy adults. Pellicle proteins preferentially adsorbed on tooth/materials were putatively associated with calcium ion homeostasis or host-microbiota interaction.


Asunto(s)
Calcio , Proteómica , Adulto , Humanos , Animales , Bovinos , Calcio/análisis , Materiales Dentales/química , Película Dental/química , Resinas Compuestas , Proteínas y Péptidos Salivales/análisis , Cementos de Ionómero Vítreo/química
7.
J Hum Evol ; 175: 103305, 2023 02.
Artículo en Inglés | MEDLINE | ID: mdl-36586354

RESUMEN

Herbivorous animals that regularly consume tannin-rich food are known to secrete certain tannin-binding salivary proteins (TBSPs), especially proline-rich proteins and histidine-rich proteins, as an effective measure to counteract the antinutritive effects of dietary tannins. Due to their high binding capacity, TBSPs complex with tannins in the oral cavity, and thereby protect dietary proteins and digestive enzymes. Although the natural diet of great apes (Hominidae) is biased toward ripe fruits, analyses of food plants revealed that their natural diet contains considerable amounts of tannins, which is raising the question of possible counter-measures to cope with dietary tannins. In our study, we investigated the salivary amino acid profiles of zoo-housed Pan paniscus, Pan troglodytes, Gorilla gorilla, and Pongo abelii, and compared their results with corresponding data from Homo sapiens. Individual saliva samples of 42 apes and 17 humans were collected and quantitated by amino acid analysis, using cation-exchange chromatography with postcolumn derivatization, following acid hydrolysis. We found species-specific differences in the salivary amino acid profiles with average total salivary protein concentration ranging from 308.8 mg/dL in Po. abelii to 1165.6 mg/dL in G. gorilla. Total salivary protein was consistently higher in ape than in human saliva samples (174 mg/dL). All apes had on average also higher relative proline levels than humans did. Histidine levels had the highest concentration in the samples from Po. abelii followed by P. paniscus. In all ape species, the high salivary concentrations of proline and histidine are considered to be indicative of high concentrations of TBSPs in hominids. Given that the species differences in salivary composition obtained in this study correspond with overall patterns of secondary compound content in the diet of wild populations, we assume that salivary composition is resilient to acute and long-lasting changes in diet composition in general and tannin content in particular.


Asunto(s)
Aminoácidos , Gorilla gorilla , Pan paniscus , Pan troglodytes , Pongo abelii , Animales , Humanos , Aminoácidos/análisis , Gorilla gorilla/metabolismo , Histidina/análisis , Pan paniscus/metabolismo , Pan troglodytes/metabolismo , Pongo abelii/metabolismo , Prolina/análisis , Saliva/química , Saliva/metabolismo , Proteínas y Péptidos Salivales/análisis , Taninos/análisis , Taninos/metabolismo , Dieta
8.
J Cancer Res Ther ; 19(Suppl 2): S669-S676, 2023 Jan 01.
Artículo en Inglés | MEDLINE | ID: mdl-38384037

RESUMEN

BACKGROUND: Radiation therapy leads to salivary gland damage that causes xerostomia, the standard radiation-induced complication during radiotherapy that affects the quality of life in head and neck cancer patients. This study was conducted at a tertiary cancer institute in Punjab state to analyze the influence of radiation therapy on various parameters and substances of saliva. MATERIALS AND METHODS: Sixty head and neck cancer patients who underwent conventional radiotherapy on a Cobalt machine were included. Saliva was collected in both stimulated and unstimulated states. Stimulated whole saliva was collected by applying two to three drops of citric acid solution (2%) over the dorsum of the tongue bilaterally at 30-s intervals for 2 min. Biochemical changes in the whole saliva were evaluated by biochemical methods at baseline, completion of therapy, and 3 and 6 months post-radiotherapy completion. RESULTS: The lowest concentration of proteins was seen after the therapy in unstimulated and stimulated saliva. Salivary protein levels showed a rising trend toward baseline in 3- and 6-month posttherapy samples. The peak value (0.4 mg/dl) was reached in the stimulated saliva after therapy. Salivary amylase did not show a consistent concentration graph. The salivary concentrations of sodium, potassium, and chloride showed peak values after radiotherapy. The lowest salivary pH was obtained at completion of therapy, both in unstimulated and stimulated saliva. After 3 months of chemoradiotherapy, the saliva reached a pH value of 8.3, whereas 6-month posttherapy sample showed a pH value of 8.4 in both unstimulated and stimulated saliva. CONCLUSIONS: At the completion of chemoradiotherapy, the total salivary protein, albumin, and inorganic components (calcium, magnesium, phosphorus) showed a downward trend from the baseline values due to the damage caused to the acinar part of the salivary gland by radiotherapy. The rise in salivary electrolytes' concentrations is attributed to the fact that even though there is loss of absorptive property of the tubular portion of the salivary gland, it retains its secretory property. Saliva becomes thick, scarce, tenacious, and acidic during the period of chemoradiotherapy.


Asunto(s)
Neoplasias de Cabeza y Cuello , Traumatismos por Radiación , Xerostomía , Humanos , Saliva/química , Calidad de Vida , Xerostomía/diagnóstico , Xerostomía/etiología , Neoplasias de Cabeza y Cuello/radioterapia , Neoplasias de Cabeza y Cuello/complicaciones , Traumatismos por Radiación/complicaciones , Quimioradioterapia/efectos adversos , Proteínas y Péptidos Salivales/análisis , Proteínas y Péptidos Salivales/metabolismo
9.
PeerJ ; 10: e14402, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-36452077

RESUMEN

Tannins are a chemical defense mechanism of plants consumed by herbivores. Variations in salivary physicochemical characteristics such as pH, total protein concentration (TP), and presence of proline-rich proteins (PRPs) in animals have been reported as a mechanism to protect the oral cavity when consuming food with variations in pH and tannins. Variations in salivary physiochemistry as adaptations for consuming tannin-rich foods have been found in omnivorous and folivorous primates, but have not yet been reported in frugivorous species such as spider monkeys. We therefore assessed changes in pH using test strips, TP concentration by measuring absorbance at 595 nm in a spectrophotometer and salivary PRPs using the SDS-PAGE electrophoresis technique in the saliva of nine captive spider monkeys in response to the consumption of solutions with different concentrations of tannic acid. The results showed variations in pH, TP concentration and the presence and variation of possible salivary PRPs associated with tannic acid concentration. These findings suggest that spider monkeys may tailor their salivary physicochemical characteristics in response to the ingestion of potentially toxic compounds.


Asunto(s)
Ateles geoffroyi , Atelinae , Animales , Saliva/química , Taninos/análisis , Proteínas y Péptidos Salivales/análisis
10.
Cold Spring Harb Protoc ; 2022(10): Pdb.top107699, 2022 10 03.
Artículo en Inglés | MEDLINE | ID: mdl-35960615

RESUMEN

Studying protein localization in mosquito salivary glands provides novel insights on the function and physiological relevance of salivary proteins and also provides an avenue to study interactions between mosquitoes and pathogens. Salivary proteins display compartmentalization. For example, proteins involved in blood feeding are stored in the medial and distal lateral lobes, whereas proteins related to sugar metabolism localize to the proximal portion of the lateral lobes. Immunohistochemistry assays use antibodies raised against recombinant salivary proteins to reveal the protein localization and interactions within the tissue. In this assay, permeabilization of the salivary glands allows the antibodies to enter the cells and bind their target proteins. The primary antibody-antigen complexes are later marked with fluorescently labeled secondary antibodies. Antibodies that recognize pathogen-specific proteins can also be incorporated in these assays, providing information about pathogen localization within the salivary glands or pathogen interactions with mosquito salivary proteins. Here, we introduce immunohistochemistry assays for use in mosquito salivary glands.


Asunto(s)
Anopheles , Animales , Inmunohistoquímica , Glándulas Salivales/química , Glándulas Salivales/metabolismo , Proteínas y Péptidos Salivales/análisis , Proteínas y Péptidos Salivales/metabolismo , Azúcares/análisis , Azúcares/metabolismo
11.
J Colloid Interface Sci ; 620: 153-167, 2022 Aug 15.
Artículo en Inglés | MEDLINE | ID: mdl-35421752

RESUMEN

The interaction of emulsions with the tongue is key to the sensory appeal of food and can potentially be exploited for oral/buccal pharmaceutical delivery. Whilst there is good understanding of the different mucoadhesive forces governing emulsion interaction with the tongue, their relative importance is not well understood. In addition, the physical location of emulsions within the saliva papillae on the tongue is not understood at all. A combination of ex vivo salivary film, and in vivo oral coating experiments were used to determine the importance of different mucoadhesive forces. Mucoadhesion of cationic emulsions was largely driven by electrostatic complexation. SDS-PAGE of the in vivo saliva coating highlighted that mucins were largely responsible for cationic emulsion mucoadhesion. Anionic emulsions were bound via hydrophobic/steric interactions to small salivary proteins typically located away from the mucin anchor points. The physical location and clustering of emulsions relative to the salivary film/papillae was probed via the invention of a fluorescent oral microscope. Cationic emulsions were densely clustered close to the papillae whilst anionic emulsions were suspended in the salivary film above the papillae. Interestingly, non-ionic emulsions were also trapped within the salivary film above the papillae as individual droplets. These findings highlight that whilst electrostatic complexation with saliva is a powerful mucoadhesive force, hydrophobic and steric interactions also act to induce oral retention of emulsions. The differences in physical location and clustering of emulsions within the salivary film hint at the 3D locations of the different salivary proteins driving each mucoadhesive interaction. This novel understanding of emulsion saliva/papillae interactions has potential to aid efficacy of buccal pharmaceutical delivery and the reduction of astringency in plant-based foods.


Asunto(s)
Boca , Proteínas y Péptidos Salivales , Emulsiones/química , Mucinas/química , Saliva/química , Proteínas y Péptidos Salivales/análisis
12.
Front Cell Infect Microbiol ; 12: 831744, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-35310855

RESUMEN

The oral cavity remains an underappreciated site for SARS-CoV-2 infection despite the myriad oral conditions observed in COVID-19 patients. Recently, replicating SARS-CoV-2 was found inside salivary epithelial cells resulting in inflammation and atrophy of salivary glands. Saliva possesses healing properties crucial for maintaining the health of the oral mucosa. Specifically, salivary antimicrobial peptides, most notable, histatin-5 exclusively produced in salivary glands, plays a vital role in innate immunity against colonizing microbial species. The demonstration of SARS-CoV-2 destruction of gland tissue where histatin-5 is produced strongly indicate that histatin-5 production is compromised due to COVID-19. Here we present a case of a patient presenting with unexplained chronic oral dysesthesia and dysgeusia post-recovery from COVID-19. To explore potential physiological mechanisms behind the symptoms, we comparatively analyzed saliva samples from the patient and matched healthy subject for histatin-5 and key cytokines. Findings demonstrated significantly reduced histatin-5 levels in patient's saliva and activation of the Th17 inflammatory pathway. As histatin-5 exhibits potent activity against the opportunistic oral pathogen Candida albicans, we evaluated saliva potency against C. albicans ex vivo. Compared to control, patient saliva exhibited significantly reduced anti-candidal efficacy. Although speculative, based on history and salivary analysis we hypothesize that salivary histatin-5 production may be compromised due to SARS-CoV-2 mediated salivary gland destruction. With the current lack of emphasis on implications of COVID-19 on oral health, this report may provide lacking mechanistic insights that may lead to reassessment of risks for oral opportunistic infections and mucosal inflammatory processes in acutely-ill and recovered COVID-19 patients.


Asunto(s)
COVID-19 , COVID-19/complicaciones , Humanos , Boca , SARS-CoV-2 , Saliva/química , Proteínas y Péptidos Salivales/análisis
13.
Mediators Inflamm ; 2021: 9198249, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34658668

RESUMEN

Allergic disease has risen to epidemic proportions since the last decade and is among the most common noncommunicable, chronic diseases in children and adolescents worldwide. Allergic disease usually occurs in early life; thus, early biomarkers of allergic susceptibility are required for preventive measures to high-risk infants which enable early interventions to decrease allergic severity. However, to date, there is no reliable general or specific allergy phenotype detection method that is easy and noninvasive for children. Most reported allergic phenotype detection methods are invasive, such as the skin prick test (SPT), oral food challenge (OFC), and blood test, and many involve not readily accessible biological samples, such as cord blood (CB), maternal blood, or newborn vernix. Saliva is a biological sample that has great potential as a biomarker measurement as it consists of an abundance of biomarkers, such as genetic material and proteins. It is easily accessible, noninvasive, collected via a painless procedure, and an easy bedside screening for real-time measurement of the ongoing human physiological system. All these advantages emphasise saliva as a very promising diagnostic candidate for the detection and monitoring of disease biomarkers, especially in children. Furthermore, protein biomarkers have the advantages as modifiable influencing factors rather than genetic and epigenetic factors that are mostly nonmodifiable factors for allergic disease susceptibility in childhood. Saliva has great potential to replace serum as a biological fluid biomarker in diagnosing clinical allergy. However, to date, saliva is not considered as an established medically acceptable biomarker. This review considers whether the saliva could be suitable biological samples for early detection of allergic risk. Such tools may be used as justification for targeted interventions in early childhood for disease prevention and assisting in reducing morbidity and mortality caused by childhood allergy.


Asunto(s)
Hipersensibilidad/diagnóstico , Saliva/química , Proteínas y Péptidos Salivales/análisis , Biomarcadores/análisis , Niño , Diagnóstico Precoz , Humanos , Hipersensibilidad/terapia , Manejo de Especímenes
14.
J Chem Ecol ; 47(8-9): 747-754, 2021 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-34550513

RESUMEN

Plant Growth-Promoting Rhizobacteria (PGPR) induce systemic resistance (SR) in plants, decreasing the development of phytopathogens. The FZB42 strain of Bacillus velezensis is known to induce an SR against pathogens in various plant species. Previous studies suggested that it could also influence the interactions between plants and associated pests. However, insects have developed several strategies to counteract plant defenses, including salivary proteins that allow the insect escaping detection, manipulating defensive pathways to its advantage, deactivating early signaling processes, or detoxifying secondary metabolites. Because Brown Marmorated Stink Bug (BMSB) Halyomorpha halys is highly invasive and polyphagous, we hypothesized that it could detect the PGPR-induced systemic defenses in the plant, and efficiently adapt its salivary compounds to counteract them. Therefore, we inoculated a beneficial rhizobacterium on Vicia faba roots and soil, previous to plant infestation with BMSB. Salivary gland proteome of BMSB was analyzed by LC-MS/MS and a label-free quantitative proteomic method. Among the differentially expressed proteins, most were up-regulated in salivary glands of insects exposed to PGPR-treated plants for 24 h. We could confirm that BMSB was confronted with a stress during feeding on PGPR-treated plants. The to-be-confirmed defensive state of the plant would have been rapidly detected by the invasive H. halys pest, which consequently modified its salivary proteins. Among the up-regulated proteins, many could be associated with a role in plant defense counteraction, and more especially in allelochemicals detoxification or sequestration.


Asunto(s)
Bacillus/crecimiento & desarrollo , Heterópteros/metabolismo , Proteínas y Péptidos Salivales/análisis , Vicia faba/microbiología , Animales , Cromatografía Líquida de Alta Presión , Heterópteros/crecimiento & desarrollo , Larva/metabolismo , Glándulas Salivales/metabolismo , Estrés Fisiológico , Espectrometría de Masas en Tándem , Regulación hacia Arriba , Vicia faba/química , Vicia faba/parasitología
15.
Tuberculosis (Edinb) ; 130: 102118, 2021 09.
Artículo en Inglés | MEDLINE | ID: mdl-34371310

RESUMEN

BACKGROUND: There is an urgent need for new, accurate, rapid, and affordable tuberculosis (TB) diagnostic tests. The aim of the present study was to use mass spectrometry to identify new preliminary candidate TB diagnostic protein biomarkers in saliva obtained from individuals with TB, and patients with other respiratory diseases (ORD). METHODS: Saliva samples were collected from 22 individuals who self-presented with symptoms suggestive of TB as part of a larger TB biomarker project. Purified salivary proteins were subjected to tryptic digestion peptides were analyzed using a QExactive Orbitrap Mass Spectrometer. Data are available via ProteomeXchange with identifier PXD027294. Identified proteins were subjected to gene ontology and ingenuity pathway analysis for functional enrichment analysis. RESULTS: 26 of the 652 identified proteins significantly discriminated individuals with TB from those with ORD after Benjamini Hochberg correction (5% FDR), with five of these proteins diagnosing TB with an AUC ≥ 0.80. A 5-protein biosignature comprising of P01011, Q8NCW5, P28072, A0A2Q2TTZ9, and Q99574 diagnosed TB with an AUC of 1.00 (95% CI, 1.00-1.00), sensitivity of 100% (95% CI, 76.2-100%) and specificity of 90.9% (95% CI, 58.7-99.8%) after leave-one-out cross validation. CONCLUSIONS: We identified novel candidate salivary protein biomarkers and biosignatures with strong potential as TB diagnostic candidates. Our results are preliminary and require validation in larger studies.


Asunto(s)
Proteínas y Péptidos Salivales/análisis , Tuberculosis/diagnóstico , Adulto , Biomarcadores/análisis , Femenino , Humanos , Masculino , Persona de Mediana Edad , Proteoma , Sensibilidad y Especificidad , Sudáfrica
16.
Parasit Vectors ; 14(1): 396, 2021 Aug 11.
Artículo en Inglés | MEDLINE | ID: mdl-34380568

RESUMEN

BACKGROUND: The argasid tick Ornithodoros moubata is the main vector in mainland Africa of African swine fever virus and the spirochete Borrelia duttoni, which causes human relapsing fever. The elimination of populations of O. moubata would contribute to the prevention and control of these two serious diseases. Anti-tick vaccines are an eco-friendly and sustainable means of eliminating tick populations. Tick saliva forms part of the tick-host interface, and knowledge of its composition is key to the identification and selection of vaccine candidate antigens. The aim of the present work is to increase the body of data on the composition of the saliva proteome of adult O. moubata ticks, particularly of females, since in-depth knowledge of the O. moubata sialome will allow the identification and selection of novel salivary antigens as targets for tick vaccines. METHODS: We analysed samples of female and male saliva using two different mass spectrometry (MS) approaches: data-dependent acquisition liquid chromatography-tandem MS (LC-MS/MS) and sequential window acquisition of all theoretical fragment ion spectra-MS (SWATH-MS). To maximise the number of proteins identified, a proteomics informed by transcriptomics analysis was applied using the O. moubata salivary transcriptomic dataset previously obtained by RNA-Seq. RESULTS: SWATH-MS proved to be superior to LC-MS/MS for the study of female saliva, since it identified 61.2% more proteins than the latter, the reproducibility of results was enhanced with its use, and it provided a quantitative picture of salivary components. In total, we identified 299 non-redundant proteins in the saliva of O. moubata, and quantified the expression of 165 of these in both male and female saliva, among which 13 were significantly overexpressed in females and 40 in males. These results indicate important quantitative differences in the saliva proteome between the sexes. CONCLUSIONS: This work expands our knowledge of the O. moubata sialome, particularly that of females, by increasing the number of identified novel salivary proteins, which have different functions at the tick-host feeding interface. This new knowledge taken together with information on the O. moubata sialotranscriptome will allow a more rational selection of salivary candidates as antigen targets for tick vaccine development.


Asunto(s)
Perfilación de la Expresión Génica , Ornithodoros/genética , Proteoma , Proteómica , Saliva/química , Proteínas y Péptidos Salivales/análisis , Animales , Proteínas de Artrópodos , Cromatografía Liquida , Femenino , Masculino , Ornithodoros/química , Reproducibilidad de los Resultados , Espectrometría de Masas en Tándem
17.
Molecules ; 26(8)2021 Apr 13.
Artículo en Inglés | MEDLINE | ID: mdl-33924512

RESUMEN

Taste plays an important role in processes such as food choices, nutrition status and health. Salivary proteins contribute to taste sensitivity. Taste reduction has been associated with obesity. Gender influences the obesity predisposition and the genetic ability to perceive the bitterness of 6-n-propylthiouracil (PROP), oral marker for food preferences and consumption. We investigated variations in the profile of salivary proteome, analyzed by HPLC-ESI-MS, between sixty-one normal weight subjects (NW) and fifty-seven subjects with obesity (OB), based on gender and PROP sensitivity. Results showed variations of taste-related salivary proteins between NW and OB, which were differently associated with gender and PROP sensitivity. High levels of Ps-1, II-2 and IB-1 proteins belonging to basic proline rich proteins (bPRPs) and PRP-1 protein belonging to acid proline rich proteins (aPRPs) were found in OB males, who showed a lower body mass index (BMI) than OB females. High levels of Ps-1 protein and Cystatin SN (Cyst SN) were found in OB non-tasters, who had lower BMI than OB super-tasters. These new insights on the role of salivary proteins as a factor driving the specific weight gain of OB females and super-tasters, suggest the use of specific proteins as a strategic tool modifying taste responses related to eating behavior.


Asunto(s)
Propiltiouracilo , Proteínas y Péptidos Salivales/análisis , Gusto/fisiología , Adulto , Anciano , Índice de Masa Corporal , Femenino , Genotipo , Humanos , Masculino , Persona de Mediana Edad , Obesidad , Adulto Joven
18.
Cancer Med ; 10(6): 2063-2074, 2021 03.
Artículo en Inglés | MEDLINE | ID: mdl-33624385

RESUMEN

AIMS: To investigate novel biomarker for diagnosis of cervical cancer, we analyzed the datasets in Gene Expression Omnibus (GEO) and confirmed the candidate biomarker in patient sample. MATERIALS AND METHODS: We collected major datasets of cervical cancer in GEO, and analyzed the differential expression of normal and cancer samples online with GEO2R and tested the differences, then focus on the GSE63514 to screen the target genes in different histological grades by using the R-Bioconductor package and R-heatmap. Then human specimens from the cervix in different histological grades were used to confirm the top 8 genes expression by immunohistochemical staining using Ki67 as a standard control. RESULTS: We identified genes differentially expressed in normal and cervical cancer, 274 upregulated genes and 206 downregulated genes. After intersection with GSE63514, we found the obvious tendency in different histological grades. Then we screened the top 24 genes, and confirmed the top 8 genes in human cervix tissues. Immunohistochemical (IHC) results confirmed that keratin 17 (KRT17) was not expressed in normal cervical tissues and was over-expressed in cervical cancer. Cysteine-rich secretory protein-2 (CRISP2) was less expressed in high-grade squamous intraepithelial lesions (HSILs) than in other histological grades. CONCLUSION: For the good repeatability and consistency of KRT17 and CRISP2, they may be good candidate biomarkers. Combined analysis of KRT17, CRISP2 expression at both genetic and protein levels can determine different histological grades of cervical squamous cell carcinoma. Such combined analysis is capable of improving diagnostic accuracy of cervical cancer.


Asunto(s)
Biomarcadores de Tumor/genética , Carcinoma de Células Escamosas/genética , Moléculas de Adhesión Celular/genética , Queratina-17/genética , Displasia del Cuello del Útero/genética , Neoplasias del Cuello Uterino/genética , Biomarcadores de Tumor/análisis , Carcinoma de Células Escamosas/química , Carcinoma de Células Escamosas/patología , Moléculas de Adhesión Celular/análisis , Proteínas de Ciclo Celular/análisis , Proteínas de Ciclo Celular/genética , Cuello del Útero/metabolismo , Proteínas de Unión al ADN/análisis , Proteínas de Unión al ADN/genética , Bases de Datos Genéticas , Conjuntos de Datos como Asunto , Desmogleína 1/análisis , Desmogleína 1/genética , Regulación hacia Abajo , Femenino , Perfilación de la Expresión Génica/métodos , Marcadores Genéticos , Humanos , Inmunohistoquímica , Péptidos y Proteínas de Señalización Intracelular/análisis , Péptidos y Proteínas de Señalización Intracelular/genética , Queratina-17/análisis , Antígeno Ki-67/genética , Antígeno Ki-67/metabolismo , Clasificación del Tumor , Proteínas de Neurofilamentos/análisis , Proteínas de Neurofilamentos/genética , Proteínas y Péptidos Salivales/análisis , Proteínas y Péptidos Salivales/genética , Proteínas de Plasma Seminal/análisis , Proteínas de Plasma Seminal/genética , Regulación hacia Arriba , Neoplasias del Cuello Uterino/química , Neoplasias del Cuello Uterino/patología , Displasia del Cuello del Útero/química , Displasia del Cuello del Útero/patología
19.
Dis Mon ; 67(9): 101165, 2021 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-33640174

RESUMEN

BACKGROUND: Saliva provides a primary defense mechanism against several infectious diseases through its numerous immunological and non-immunological factors. Alteration in the composition of saliva often compromises its defense mechanisms, predisposing the oral cavity to disease entities. HIV patients under antiretroviral therapy (ART) have shown to exhibit altered salivary composition. These changes are postulated to be a result of the effect of ART on the salivary protein and electrolytes levels. OBJECTIVES: The present study aims to assess the potential difference in the salivary total protein and electrolyte levels in HIV patients with and without ART. METHODS: Patients were divided into 3 groups- Group A (HIV-1 positive patient under ART for at least 6 months)-66, Group B (HIV-1 positive patient not started on ART)-66, Group C (HIV negative patients)-66. Saliva samples were collected and evaluated for total salivary protein and electrolyte levels in all the 3 groups. RESULTS: There was a statistically significant difference in the salivary protein (p = 0.000) and electrolyte (Sodium, p = 0.000; Potassium, p = 0.039; chlorine, p = 0.027; ionized calcium, p = 0.002) levels among the three groups. CONCLUSION: HIV positive individuals with and without ART have alteration in the salivary composition. Some of these alterations (total protein and iCa levels) are due to the HIV infection, while others (Na, K, Cl) could be due to ART or a combined effect of both. Salivary changes in HIV positive individuals could predispose them to oral diseases. Thus, regular oral examination and prophylactic regimen must be formulated to maintain their oral hygiene and quality of life.


Asunto(s)
Fármacos Anti-VIH/efectos adversos , Terapia Antirretroviral Altamente Activa/efectos adversos , Electrólitos/análisis , Infecciones por VIH , Saliva/efectos de los fármacos , Proteínas y Péptidos Salivales/análisis , Adulto , Fármacos Anti-VIH/uso terapéutico , Calcio/metabolismo , Cloro/metabolismo , Femenino , Infecciones por VIH/complicaciones , Infecciones por VIH/tratamiento farmacológico , Humanos , Masculino , Persona de Mediana Edad , Enfermedades de la Boca , Potasio/metabolismo , Calidad de Vida , Saliva/química
20.
Medicine (Baltimore) ; 100(6): e24539, 2021 Feb 12.
Artículo en Inglés | MEDLINE | ID: mdl-33578547

RESUMEN

ABSTRACT: The purpose of the present study was to investigate the accuracy of a screening method using salivary tests to screen for periodontal disease.In total, 1888 individuals older than 30 years in 2017 and 2296 in 2018 who underwent medical check-ups for metabolic syndrome agreed to participate and simultaneously underwent a dental examination by dentists and salivary tests. Salivary occult blood, protein, and ammonia levels and white blood cell counts were evaluated in salivary tests using commercially available kits. The relationship between the results of the salivary tests and dental examination was examined and classification performance was analyzed.The prevalence of periodontal disease was 69.9% in 2017 and 66.8% in 2018. Salivary ammonia showed the highest classification performance in both years (sensitivity 83.5 and 83.1%, precision 73.0 and 69.3%, F-measure 0.779 and 0.756). Occult blood, which was assessed using a monoclonal antibody to human hemoglobin, also showed good performance (sensitivity 69.5%, precision 70.6%, F-measure 0.701). Questions regarding self-reported gingival bleeding were not sufficient to screen for periodontitis. The present results suggest that screening tests using salivary samples may detect periodontal disease in approximately 70% to 80% of subjects in a large population.Conclusion: Salivary ammonia and hemoglobin have potential as salivary markers in the screening of periodontal disease.


Asunto(s)
Tamizaje Masivo , Enfermedades Periodontales/diagnóstico , Saliva/química , Anciano , Amoníaco/análisis , Femenino , Humanos , Japón , Recuento de Leucocitos , Masculino , Tamizaje Masivo/métodos , Persona de Mediana Edad , Sangre Oculta , Examen Físico/métodos , Juego de Reactivos para Diagnóstico , Reproducibilidad de los Resultados , Proteínas y Péptidos Salivales/análisis , Sensibilidad y Especificidad
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