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Zhang S, Clasen F, Cai H, Do T, Shoaie S, Carpenter GH. Nitrate supplementation affects taste by changing the oral metabolome and microbiome. NPJ Biofilms Microbiomes 2025; 11:69. [PMID: 40316518 PMCID: PMC12048645 DOI: 10.1038/s41522-025-00689-8] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/16/2024] [Accepted: 04/01/2025] [Indexed: 05/04/2025] Open
Abstract
Nitrate, an inorganic anion found in various foods is also present in saliva and has emerged as a potential prebiotic for the oral microbiome. Salivary glands concentrate nitrate from the bloodstream and release it into the oral cavity via the anion transporter sialin SLC17A5. In previous studies dietary nitrate supplementation altered oral bacteria composition, favouring genera like Rothia and Neisseria while reducing Streptococcus, Veillonella, Prevotella, and Actinomyces. The present study hypothesized that taste intensity might adapt to changes in the oral microbiome caused by nitrate supplementation. Participants underwent taste tests before, during, and after supplementation. All subjects showed greater levels of salivary nitrate during supplementation and had higher levels of Neisseria compared to before. Subjects were then grouped according to taste tests (before vs. during) as responders (ANOVA p < 0.05, n = 7), and non-responders (ANOVA p > 0.05, n = 6) and their salivary metabolome and oral microbiome further analysed. Responders had significantly less 5-amino pentanoate, formate, propionate and butyrate in saliva while non-responders showed no metabolite changes between before and during supplementation. In contrast, non-responders had increased Capnocytophaga gingivalis and altered lysosomal degradation pathways. Overall, nitrate supplementation shifted the oral microbiome composition in all subjects and when taste intensity was altered this correlated to bacteria-derived short-chain fatty acid production. This suggests taste perception is affected by the oral microbiome.
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Affiliation(s)
- Shuyuan Zhang
- Centre for Host Microbiome Interactions, Faculty of Dentistry, Oral and Craniofacial Sciences, King's College London, London, UK.
| | - Frederick Clasen
- Centre for Host Microbiome Interactions, Faculty of Dentistry, Oral and Craniofacial Sciences, King's College London, London, UK
| | - Haizhuang Cai
- Centre for Host Microbiome Interactions, Faculty of Dentistry, Oral and Craniofacial Sciences, King's College London, London, UK
| | - Thuy Do
- School of Dentistry, Faculty of Medicine and Health, University of Leeds, LS2 9LU, Leeds, UK
| | - Saeed Shoaie
- Centre for Host Microbiome Interactions, Faculty of Dentistry, Oral and Craniofacial Sciences, King's College London, London, UK
| | - Guy H Carpenter
- Centre for Host Microbiome Interactions, Faculty of Dentistry, Oral and Craniofacial Sciences, King's College London, London, UK.
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Albahri J, Allison H, Whitehead KA, Muhamadali H. The role of salivary metabolomics in chronic periodontitis: bridging oral and systemic diseases. Metabolomics 2025; 21:24. [PMID: 39920480 PMCID: PMC11805826 DOI: 10.1007/s11306-024-02220-0] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 07/24/2024] [Accepted: 12/31/2024] [Indexed: 02/09/2025]
Abstract
BACKGROUND Chronic periodontitis is a condition impacting approximately 50% of the world's population. As chronic periodontitis progresses, the bacteria in the oral cavity change resulting in new microbial interactions which in turn influence metabolite production. Chronic periodontitis manifests with inflammation of the periodontal tissues, which is progressively developed due to bacterial infection and prolonged bacterial interaction with the host immune response. The bi-directional relationship between periodontitis and systemic diseases has been reported in many previous studies. Traditional diagnostic methods for chronic periodontitis and systemic diseases such as chronic kidney diseases (CKD) have limitations due to their invasiveness, requiring practised individuals for sample collection, frequent blood collection, and long waiting times for the results. More rapid methods are required to detect such systemic diseases, however, the metabolic profiles of the oral cavity first need to be determined. AIM OF REVIEW In this review, we explored metabolomics studies that have investigated salivary metabolic profiles associated with chronic periodontitis and systemic illnesses including CKD, oral cancer, Alzheimer's disease, Parkinsons's disease, and diabetes to highlight the most recent methodologies that have been applied in this field. KEY SCIENTIFIC CONCEPTS OF THE REVIEW Of the rapid, high throughput techniques for metabolite profiling, Nuclear magnetic resonance (NMR) spectroscopy was the most applied technique, followed by liquid chromatography-mass spectrometry (LC-MS) and gas chromatography-mass spectrometry (GC-MS). Furthermore, Raman spectroscopy was the most used vibrational spectroscopic technique for comparison of the saliva from periodontitis patients to healthy individuals, whilst Fourier Transform Infra-Red Spectroscopy (FT-IR) was not utilised as much in this field. A recommendation for cultivating periodontal bacteria in a synthetic medium designed to replicate the conditions and composition of saliva in the oral environment is suggested to facilitate the identification of their metabolites. This approach is instrumental in assessing the potential of these metabolites as biomarkers for systemic illnesses.
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Affiliation(s)
- Jawaher Albahri
- Centre for Metabolomics Research, Department of Biochemistry, Cell and Systems Biology, Institute of Systems, Molecular and Integrative Biology, University of Liverpool, Liverpool, L69 7ZB, UK
- Department of Pharmaceutical Chemistry, College of Pharmacy, King Khalid University, Abha, 62529, Saudi Arabia
| | - Heather Allison
- Institute of Infection, Veterinary and Ecological Sciences, University of Liverpool, Liverpool, UK
| | - Kathryn A Whitehead
- Microbiology at Interfaces, Department of Life Sciences, Manchester Metropolitan University, Chester St, Manchester, M1 5GD, UK.
| | - Howbeer Muhamadali
- Centre for Metabolomics Research, Department of Biochemistry, Cell and Systems Biology, Institute of Systems, Molecular and Integrative Biology, University of Liverpool, Liverpool, L69 7ZB, UK.
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Rovera A, Alzahrani AS, Rovera G, Anderson P. Influence of Parotid Saliva Composition on Phosphate Species' Chemical State in Relation to Dental Caries. Clin Exp Dent Res 2025; 11:e70089. [PMID: 39940089 PMCID: PMC11821548 DOI: 10.1002/cre2.70089] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/01/2024] [Revised: 01/19/2025] [Accepted: 01/28/2025] [Indexed: 02/14/2025] Open
Abstract
OBJECTIVES The aim of the present study was to analyze the influence of parotid saliva (PS) composition on the phosphate species' chemical state in relation to dental caries. METHODS Unilateral stimulated PS samples were collected from 21 healthy adult subjects using a Lashley cup. Clinical caries scores of each subject were classified using the ICDAS score. The pH was recorded for each saliva sample. The concentration of specific inorganic elements (K+, Na+, Ca2+, Al3+, Sr2+, Li+, Zn2+, and Mg2+) was measured using an Inductively Coupled Plasma-Optical Emission Spectrometer (ICP-OES). The PS effective concentration of ions and the degree of saturation (DS) with respect to enamel mineral were determined by speciation calculation. The PS chemical environment was determined using 31P-Nuclear-Magnetic-Resonance-Spectroscopy (31P-NMR). Pearson's correlation test was applied to evaluate the influence of PS composition on the ICDAS score. RESULTS The pH varied from 5.9 to 7.6. The ICDAS scores varied from 0.03 to 2.48. PS was supersaturated with respect to HAp at all pH values. The 31P-NMR peak position value reflects the phosphate chemical state within PS and its change in relation to pH. Only calcium ion concentration [Ca2+], aluminum activity {Al3+}, and DSHAp significantly correlated with the 31P-NMR peak position value (ppm), whereas no correlation was observed between the 31P-NMR peak position value and the activities of K+, Na+, Sr2+, Mg2+, Li+, and Zn2+. CONCLUSIONS This parotid saliva 31P-NMR study has shown that calcium ion concentration [Ca2+], aluminum activity {Al3+}, and DSHAp significantly influence the phosphate species' chemical state existing within PS and provides extended knowledge on the main biochemical determinants of the caries process.
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Affiliation(s)
- Angela Rovera
- Dental Physical Sciences Unit, Centre for Oral Bioengineering, Institute of DentistryQueen Mary University of LondonLondonUK
| | - Ali S. Alzahrani
- Department of Basic Medical, Oral and Allied Dental Sciences, College of DentistryTaif UniversityTaifSaudi Arabia
| | - Guido Rovera
- Nuclear Medicine Division, Department of Medical SciencesUniversity of TurinTurinItaly
| | - Paul Anderson
- Dental Physical Sciences Unit, Centre for Oral Bioengineering, Institute of DentistryQueen Mary University of LondonLondonUK
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Nakhod VI, Butkova TV, Malsagova KA, Petrovskiy DV, Izotov AA, Nikolsky KS, Kaysheva AL. Sample Preparation for Metabolomic Analysis in Exercise Physiology. Biomolecules 2024; 14:1561. [PMID: 39766268 PMCID: PMC11673972 DOI: 10.3390/biom14121561] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/03/2024] [Revised: 11/22/2024] [Accepted: 11/25/2024] [Indexed: 01/11/2025] Open
Abstract
Metabolomics investigates final and intermediate metabolic products in cells. Assessment of the human metabolome relies principally on the analysis of blood, urine, saliva, sweat, and feces. Tissue biopsy is employed less frequently. Understanding the metabolite composition of biosamples from athletes can significantly improve our knowledge of molecular processes associated with the efficiency of training and recovery. Such knowledge may also lead to new management opportunities. Successful execution of metabolomic studies requires simultaneous qualitative and quantitative analyses of numerous small biomolecules in samples under test. Unlike genomics and proteomics, which do not allow for direct assessment of enzymatic activity, metabolomics focuses on biochemical phenotypes, providing unique information about health and physiological features. Crucial factors in ensuring the efficacy of metabolomic analysis are the meticulous selection and pre-treatment of samples.
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Affiliation(s)
| | | | - Kristina A. Malsagova
- Institute of Biomedical Chemistry, 109028 Moscow, Russia; (V.I.N.); (T.V.B.); (D.V.P.); (A.A.I.); (K.S.N.); (A.L.K.)
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Catussi BLC, Lo Turco EG, Pereira DM, Teixeira RMN, Castro BP, Massaia IFD. Metabolomics: Unveiling biological matrices in precision nutrition and health. Clin Nutr ESPEN 2024; 64:314-323. [PMID: 39427750 DOI: 10.1016/j.clnesp.2024.10.148] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/11/2024] [Revised: 10/07/2024] [Accepted: 10/14/2024] [Indexed: 10/22/2024]
Abstract
Precision nutrition, an expanding field at the intersection of nutrition science and personalized medicine, is rapidly evolving with metabolomics integration. Metabolomics, facilitated by advanced technologies like mass spectrometry (MS) and nuclear magnetic resonance (NMR) spectroscopy, facilitates comprehensive profiling of metabolites across diverse biological samples. From the perspective of health care systems, precision nutrition gains relevance due to the substantial impact of prevalent non-communicable diseases (NCDs) on societal well-being, which is directly linked with dietary habits and eating behavior. Furthermore, biomarker products derived from metabolomics have been utilized in Europe, the USA, and Brazil to understand metabolic dysregulations and tailor diets accordingly. Despite its burgeoning status, metabolomics holds great potential in revolutionizing nutritional science, particularly with the integration of artificial intelligence and machine learning, offering novel insights into personalized dietary interventions and disease prediction. This narrative review emphasizes the transformative impact of metabolomics in precision and delineates avenues for future research and application, paving the way for a more tailored and practical approach to nutrition management.
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Garcia PN, de Souza MM, Izidoro MA, Juliano L, Lourenço SV, Camillo CMC. Saliva metabolomics: concepts and applications in oral disorders. Clin Oral Investig 2024; 28:579. [PMID: 39377832 DOI: 10.1007/s00784-024-05990-y] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/12/2024] [Accepted: 10/02/2024] [Indexed: 10/09/2024]
Abstract
OBJECTIVES The purpose of this review was to present the basic concepts of metabolomics methodology and the use of saliva for diagnostic, prognostic, and predictive strategies. MATERIAL AND METHODS This review followed the focus in: "saliva metabolomics" and "oral diseases". The authors searched studies on PubMed database. The inclusion criteria were original studies and reviews that assessed metabolomics techniques. A descriptive analysis was performed considering the study design, approach system, clinical steps, and tools for the determination of profile or biomarkers metabolites, and the advantages and disadvantages. RESULTS Metabolomic analyses use a combination of analytical instrumentation and informatic tools to provide information on metabolite characteristics. In this review we described different technologies applied and the advantages and limitations of each technique. Furthermore, in the literature search, we retrieved 25 studies that investigated saliva metabolites in oral diseases: 8 studies used targeted analysis and 17 untargeted metabolomics approaches. Most studies included patients with periodontal diseases, oral squamous cell carcinoma, and Sjögren Syndrome. The most frequently reported metabolites were glycine, leucine, phenylalanine, dipeptides, linoleic acid, arachidonic acid, tyrosine, choline, taurine, lactate, valine, and proline. CONCLUSIONS Metabolomics analysis has emerged as a powerful tool for tumor diagnosis and to enhance tumor classification, including salivary gland tumors (SGTs). It also holds promise for developing personalized treatment plans and defining more precise prognostic categories. CLINICAL RELEVANCE Metabolomics is the most functional and comprehensive technique for monitoring and understanding gene functions and identifying the biochemical state of an organism in response to genetic and environmental changes.
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Affiliation(s)
- Pedro Nunes Garcia
- International Research Center, Centro Internacional de Pesquisa, A.C. Camargo Cancer Center, Rua Taguá, 440 - Primeiro andar, São Paulo, 01508-010, Brazil
| | - Milena Monteiro de Souza
- International Research Center, Centro Internacional de Pesquisa, A.C. Camargo Cancer Center, Rua Taguá, 440 - Primeiro andar, São Paulo, 01508-010, Brazil.
| | | | - Luiz Juliano
- Department of Biophysics, Federal University of São Paulo, São Paulo, Brazil
| | | | - Cláudia Malheiros Coutinho Camillo
- International Research Center, Centro Internacional de Pesquisa, A.C. Camargo Cancer Center, Rua Taguá, 440 - Primeiro andar, São Paulo, 01508-010, Brazil
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Alkahtani A, Grootveld M, Bhogadia M, Baysan A. Exploring Salivary Metabolic Alterations in Type 2 Diabetes: Implications for Dental Caries and Potential Influences of HbA1c and Vitamin D Levels. Metabolites 2024; 14:372. [PMID: 39057695 PMCID: PMC11279097 DOI: 10.3390/metabo14070372] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/25/2024] [Revised: 06/16/2024] [Accepted: 06/25/2024] [Indexed: 07/28/2024] Open
Abstract
Diabetes mellitus is considered to be the most common health issue affecting almost 1 in 11 adults globally. Oral health complications including xerostomia, periodontal disease, dental caries, and soft tissue lesions are prevalent among individuals with diabetes, and therefore an understanding of the potential association between salivary metabolites and dental caries progression would enable the early detection and prevention of this non-communicable disease. Therefore, the aim of this study was to compare salivary biomarkers between individuals with type 2 diabetes (T2DM) with those without this disorder (ND) using 1H NMR-based metabolomics strategies. The objectives were to identify T2DM-associated biomarker signatures and their potential impact on dental caries. In addition, HbA1c and vitamin D levels were also analysed for this purpose. METHODS Stimulated whole-mouth saliva (SWS) samples were collected from T2DM and ND (n = 30 in each case) participants randomly selected from a group of 128 participants recruited for this case-control study. All participants were advised to refrain from eating, drinking, and smoking for at least 1-2 h prior to sample collection. Following preparation, SWS supernatants underwent 1H NMR analysis at an operating frequency of 800 MHz, and the dataset acquired was analysed using a range of multivariate metabolomics techniques. RESULTS Metabolomics analysis of data acquired demonstrated that, together with up- and downregulated blood HbA1c and vitamin D levels, key salivary discriminators between these two classifications included lactate, taurine, creatinine, α-glucose, and formate to a lesser extent. The bacterial catabolites lactate and formate were both significantly upregulated in the T2DM group, and these have previously been implicated in the pathogenesis of dental caries. Significance analysis of metabolites (SAM)-facilitated AUROC analysis yielded an 83% accuracy for this distinction. CONCLUSION In conclusion, this study highlights the significant differences in salivary metabolites between individuals with T2DM and healthy controls. Such differences appear to be related to the development and progression of dental caries in T2DM patients.
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Affiliation(s)
- Ashwaq Alkahtani
- Institute of Dentistry, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, London E1 2AD, UK;
| | - Martin Grootveld
- Leicester School of Pharmacy, De Montfort University, The Gateway, Leicester LE1 9BH, UK; (M.B.); (M.G.)
| | - Mohammed Bhogadia
- Leicester School of Pharmacy, De Montfort University, The Gateway, Leicester LE1 9BH, UK; (M.B.); (M.G.)
| | - Aylin Baysan
- Institute of Dentistry, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, London E1 2AD, UK;
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Antonelli R, Ferrari E, Gallo M, Ciociola T, Calciolari E, Spisni A, Meleti M, Pertinhez TA. The Association between Salivary Metabolites and Gingival Bleeding Score in Healthy Subjects: A Pilot Study. Int J Mol Sci 2024; 25:5448. [PMID: 38791486 PMCID: PMC11122368 DOI: 10.3390/ijms25105448] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/13/2024] [Revised: 05/06/2024] [Accepted: 05/15/2024] [Indexed: 05/26/2024] Open
Abstract
Periodontal diseases, including gingivitis and periodontitis, are among the most prevalent diseases in humans. Gingivitis is the mildest form of periodontal disease, characterized by inflammation of the gingiva caused by the accumulation of dental plaque. Salivary diagnostics are becoming increasingly popular due to the variation in saliva composition in response to pathological processes. We used a metabolomics approach to investigate whether a specific saliva metabolic composition could indicate preclinical stage of gingivitis. 1H-NMR spectroscopy was used to obtain the salivary metabolite profiles of 20 healthy subjects. Univariate/multivariate statistical analysis evaluated the whole saliva metabolite composition, and the Full-Mouth Bleeding Score (FMBS) was employed as a classification parameter. Identifying a signature of specific salivary metabolites could distinguish the subjects with high FMBS scores but still within the normal range. This set of metabolites may be due to the enzymatic activities of oral bacteria and be associated with the early stages of gingival inflammation. Although this analysis is to be considered exploratory, it seems feasible to establish an FMBS threshold that distinguishes between the absence and presence of early inflammatory alterations at the salivary level.
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Affiliation(s)
- Rita Antonelli
- Department of Medicine and Surgery, Centro Universitario di Odontoiatria, University of Parma, 43126 Parma, Italy; (R.A.); (E.C.); (M.M.)
| | - Elena Ferrari
- Laboratory of Biochemistry and Metabolomics, Department of Medicine and Surgery, University of Parma, 43125 Parma, Italy; (E.F.); (M.G.); (T.A.P.)
| | - Mariana Gallo
- Laboratory of Biochemistry and Metabolomics, Department of Medicine and Surgery, University of Parma, 43125 Parma, Italy; (E.F.); (M.G.); (T.A.P.)
| | - Tecla Ciociola
- Laboratory of Microbiology and Virology, Department of Medicine and Surgery, University of Parma, 43125 Parma, Italy;
| | - Elena Calciolari
- Department of Medicine and Surgery, Centro Universitario di Odontoiatria, University of Parma, 43126 Parma, Italy; (R.A.); (E.C.); (M.M.)
- Center for Oral Clinical Research, Institute of Dentistry, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, London E1 4NS, UK
| | - Alberto Spisni
- Laboratory of Biochemistry and Metabolomics, Department of Medicine and Surgery, University of Parma, 43125 Parma, Italy; (E.F.); (M.G.); (T.A.P.)
| | - Marco Meleti
- Department of Medicine and Surgery, Centro Universitario di Odontoiatria, University of Parma, 43126 Parma, Italy; (R.A.); (E.C.); (M.M.)
| | - Thelma A. Pertinhez
- Laboratory of Biochemistry and Metabolomics, Department of Medicine and Surgery, University of Parma, 43125 Parma, Italy; (E.F.); (M.G.); (T.A.P.)
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Fang L, Zhai Q, Zhang H, Ji P, Chen C, Zhang H. Comparisons of different extraction methods and solvents for saliva samples. Metabolomics 2024; 20:38. [PMID: 38460055 DOI: 10.1007/s11306-024-02105-2] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 02/27/2023] [Accepted: 02/19/2024] [Indexed: 03/11/2024]
Abstract
INTRODUCTION Changes in the categories and concentrations of salivary metabolites may be closely related to oral, intestinal or systemic diseases. To study salivary metabolites, the first analytical step is to extract them from saliva samples as much as possible, while reducing interferences to a minimum. Frequently used extraction methods are protein precipitation (PPT), liquid-liquid extraction (LLE) and solid-phase extraction (SPE), with various organic solvents. The types and quantities of metabolites extracted with different methods may vary greatly, but few studies have systematically evaluated them. OBJECTIVES This study aimed to select the most suitable methods and solvents for the extraction of saliva according to different analytical targets. METHODS An untargeted metabolomics approach based on liquid chromatography-mass spectrometry was applied to obtain the raw data. The numbers of metabolites, repeatability of the data and intensities of mass spectrometry signals were used as evaluation criteria. RESULTS PPT resulted in the highest coverage. Among the PPT solvents, acetonitrile displayed the best repeatability and the highest coverage, while acetone resulted in the best signal intensities for the extracted compounds. LLE with the mixture of chloroform and methanol was the most suitable for the extraction of small hydrophobic compounds. CONCLUSION PPT with acetonitrile or acetone was recommended for untargeted analysis, while LLE with the mixture of chloroform and methanol was recommended for small hydrophobic compounds.
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Affiliation(s)
- Lingli Fang
- Chongqing Key Laboratory of Oral Disease and Biomedical Sciences, Stomatological Hospital of Chongqing Medical University, Chongqing, China
| | - Qiming Zhai
- Chongqing Key Laboratory of Oral Disease and Biomedical Sciences, Stomatological Hospital of Chongqing Medical University, Chongqing, China
| | - Hua Zhang
- Department of Obstetrics, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China
| | - Ping Ji
- Chongqing Key Laboratory of Oral Disease and Biomedical Sciences, Stomatological Hospital of Chongqing Medical University, Chongqing, China
| | - Chang Chen
- Institute of Life Sciences, Chongqing Medical University, Chongqing, China.
| | - Hongmei Zhang
- Department of Pediatric Dentistry, Stomatological Hospital of Chongqing Medical University, Chongqing, China.
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Rovera A, Anderson P. Influence of gender and combined estrogen-progestin oral contraceptive on parotid saliva flow rate, pH, and electrolytes concentration. Clin Exp Dent Res 2024; 10:e800. [PMID: 37933471 PMCID: PMC10860451 DOI: 10.1002/cre2.800] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/14/2023] [Revised: 10/02/2023] [Accepted: 10/07/2023] [Indexed: 11/08/2023] Open
Abstract
OBJECTIVES Endocrinal variations within an individual impact the electrolyte composition, pH, and flow-rate (FR) of saliva. The aim of this study was to evaluate the gender-specific differences and the effect of combined estrogen-progestin oral contraceptives (COCs) on FR, pH, and electrolyte concentrations in the parotid saliva (PS) of a group of healthy adults. MATERIAL AND METHODS Stimulated PS was collected from 20 healthy adults using a Lashley cup; 11 males, 8 females, and 1 female undertaking combined contraceptive therapy (levonorgestrel/ethinyloestradiol 0.1 mg + 0.02 mg). FR and pH were recorded for each saliva sample. Electrolytes concentrations (Na+ , Ca2+ , K+ , Mg2+ ) were measured using inductively coupled plasma optical emission spectrometer (ICP-OES). Statistical analysis was performed, and the significance level was set at p < .05. RESULTS PS FR varied from 0.13 to 0.42 mL/min in females not taking any medication and from 0.08 to 0.5 mL/min in males not taking any medication. PS pH of females and males not taking any medication ranged from 6.23 to 7.50 and from 6.15 to 7.55. PS pH and FR of the female taking COCs were 6.5 and 0.1 mL/min. PS pH, FR, and electrolytes concentrations (Ca2+ , Na+ , K+ , Mg2+ ) were not statistically significantly different between females and males not taking any medication. PS concentrations of Ca2+ and Na+ were significantly higher in the females taking COCs than in the females not taking any medication. Whereas, concentrations of K+ and Mg2+ did not differ significantly between the females taking COCs and the females not taking any medication. CONCLUSIONS There are no significant gender-specific differences in PS flow rate, pH, and electrolyte concentrations of Na+ , Ca2+ , Mg2+ , and K+ . Combined hormonal oral contraceptive has a significant effect on PS flow rate, pH, Ca2+ , and Na+ concentrations. Whereas the PS concentration of K+ and Mg2+ are not influenced by COCs. These results warrant further investigation.
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Affiliation(s)
- Angela Rovera
- Dental Physical Sciences Unit, Centre for Oral Bioengineering, Institute of DentistryQueen Mary University of LondonLondonUK
| | - Paul Anderson
- Dental Physical Sciences Unit, Centre for Oral Bioengineering, Institute of DentistryQueen Mary University of LondonLondonUK
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Zheng L, Shi L, Wu X, Hu P, Zhang B, Han X, Wang K, Li X, Yang F, Wang Y, Li X, Qiao R. Advances in Research on Pig Salivary Analytes: A Window to Reveal Pig Health and Physiological Status. Animals (Basel) 2024; 14:374. [PMID: 38338017 PMCID: PMC10854898 DOI: 10.3390/ani14030374] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/12/2023] [Revised: 01/17/2024] [Accepted: 01/22/2024] [Indexed: 02/12/2024] Open
Abstract
Saliva is an important exocrine fluid that is easy to collect and is a complex mixture of proteins and other molecules from multiple sources from which considerable biological information can be mined. Pig saliva, as an easily available biological liquid rich in bioactive ingredients, is rich in nucleic acid analytes, such as eggs, enzymes, amino acids, sugars, etc. The expression levels of these components in different diseases have received extensive attention, and the analysis of specific proteins, metabolites, and biological compositions in pig saliva has become a new direction for disease diagnosis and treatment. The study of the changes in analytes in pig saliva can provide a new strategy for early diagnosis, prognosis assessment, and treatment of diseases. In this paper, the detection methods and research progress of porcine salivary analytes are reviewed, the application and research progress of porcine salivary analytes in diseases are discussed, and the future application prospect is presented.
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Affiliation(s)
| | | | | | | | | | | | | | | | | | | | | | - Ruimin Qiao
- College of Animal Science and Technology, Henan Agricultural University, Zhengzhou 450046, China; (L.Z.); (L.S.)
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Musalem-Dominguez O, Montiel-Company JM, Ausina-Márquez V, Morales-Tatay JM, Almerich-Silla JM. Salivary metabolomic profile associated with cariogenic risk in children. J Dent 2023; 136:104645. [PMID: 37524196 DOI: 10.1016/j.jdent.2023.104645] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/24/2023] [Revised: 07/27/2023] [Accepted: 07/28/2023] [Indexed: 08/02/2023] Open
Abstract
OBJECTIVES To identify the metabolomic differences in the saliva of healthy children versus children with active carious lesions and to estimate the predictive capacity of a model based on the salivary metabolomic profile. METHODS A study of cases (n = 31) and controls (n = 37) was designed for children aged between 6 and 12 (mean age of the cases: 8.9; controls: 8.7). The said children attended public health centers in Valencia, Spain. Intraoral examinations were performed by a single examiner using ICDAS II diagnostic criteria. Unstimulated total saliva samples were analyzed by nuclear magnetic resonance (NMR) spectroscopy. RESULTS The dft index for cases was 2.84 while it was 0.19 for the control group, the DMFT index was 1.13 and 0.11, respectively. The predictive model generated by the multivariate PLS-DA analysis projects a separation between the cases and the controls on the score chart with a predictive capacity and generating an area under the curve of 0.71. The metabolites: 3-methyl-2-oxovalerate, 3-hydroxybutyrate, lactate, acetone, citrate, ornithine, ethanolamine, taurine, proline, glycine, mannose, glucose, 1-6-Anhydro-β-d-glucose and citraconate, are those that show greater significance in the model. In the controls, glycine (Cohen's d = 0.430) and glucose (Cohen's d = 0.560) present higher means compared to the cases. On the contrary, taurine (Cohen's d= -0.474) and mannose (Cohen's d= -0.456) show higher means in cases compared to controls. CONCLUSIONS Our findings show a difference in the salivary metabolomic profiles, specifically in the groups of saccharides and amino acids, suggesting an association of these with the level of caries risk. CLINICAL SIGNIFICANCE The results reported in the present study reinforce the use of salivary metabolomics as a research method for the search for salivary biomarkers that allow the evaluation of caries risk in patients. Furthermore, it brings us closer to a personalized medicine that will help in dental caries prevention strategies.
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Affiliation(s)
- Oscar Musalem-Dominguez
- Departament d`Estomatologia, Facultat de Medicina i Odontologia, Universitat de Valencia, Gascó Oliag, 1, Valencia 46010, Spain
| | - José María Montiel-Company
- Departament d`Estomatologia, Facultat de Medicina i Odontologia, Universitat de Valencia, Gascó Oliag, 1, Valencia 46010, Spain.
| | - Verónica Ausina-Márquez
- Department of Dentistry, European University of Valencia, Passeig de l'Albereda, 7, Valencia 46010, Spain
| | - José Manuel Morales-Tatay
- Department of Pathology, Medicine and Odontology Faculty, University of Valencia, Valencia 46010, Spain; Research Unit, INCLIVA Health Research Institute, Valencia 46010, Spain
| | - José Manuel Almerich-Silla
- Departament d`Estomatologia, Facultat de Medicina i Odontologia, Universitat de Valencia, Gascó Oliag, 1, Valencia 46010, Spain
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Foratori-Junior GA, Le Guennec A, Fidalgo TKDS, Jarvis J, Mosquim V, Buzalaf MAR, Carpenter GH, Sales-Peres SHDC. Comparison of the Metabolic Profile between Unstimulated and Stimulated Saliva Samples from Pregnant Women with/without Obesity and Periodontitis. J Pers Med 2023; 13:1123. [PMID: 37511736 PMCID: PMC10381358 DOI: 10.3390/jpm13071123] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/16/2023] [Revised: 07/08/2023] [Accepted: 07/09/2023] [Indexed: 07/30/2023] Open
Abstract
This study aimed to compare the metabolic profile of unstimulated (US) and stimulated (SS) saliva samples from pregnant women with/without obesity and periodontitis. Ninety-six pregnant women were divided into: obesity + periodontitis (OP = 20); obesity/no periodontitis (OWP = 27); normal BMI + periodontitis (NP = 20); and normal BMI/no periodontitis (NWP = 29). US and SS samples were collected by expectoration and chewing of sterilized parafilm gum, respectively, and samples were individually analyzed by Proton Nuclear Magnetic Resonance (1H-NMR). Univariate (t test and correlations) and multivariate (Principal Component Analysis-PCA, and Partial Least Square-Discriminant Analysis-PLS-DA with Variance Importance Projection-VIP scores) and Metabolite Set Enrichment Analysis were done (p < 0.05). Metabolites commonly found in all groups in elevated concentration in US samples were 5-Aminopentoate, Acetic acid, Butyric acid, Propionic acid, Pyruvic acid, and Succinic acid. They were mainly related to the butyrate metabolism, citric acid cycle, amino sugar metabolism, fatty acids biosynthesis, pyruvate metabolism, glutamate metabolism, and Warburg effect. Metabolites commonly found in all groups that were in elevated concentration in SS samples were Citrulline, Fumaric acid, Histidine, N-acetyl glutamine, N-acetylneuraminic acid, para-hydroxyphenylacetic acid, Proline, Tyrosine. Although some differences were found between unstimulated and stimulated saliva samples from pregnant women with/without obesity and periodontitis, stimulated saliva collection seems adequate, demonstrating similar metabolic pathways to unstimulated saliva samples when groups are compared.
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Affiliation(s)
- Gerson Aparecido Foratori-Junior
- Department of Pediatric Dentistry, Orthodontics and Public Health, Bauru School of Dentistry, University of São Paulo, Bauru 17012-901, Brazil
| | - Adrien Le Guennec
- Nuclear Magnetic Resonance Facility, Guy’s Campus, King’s College London, London SE1 1UL, UK
| | - Tatiana Kelly da Silva Fidalgo
- Department of Preventive and Community Dentistry, School of Dentistry, Rio de Janeiro State University, Rio de Janeiro 20551-030, Brazil
| | - James Jarvis
- Randall Division of Cell and Molecular Biophysics and Centre for Biomolecular Spectroscopy, Guy’s Campus, King’s College London, London SE1 1UL, UK
| | - Victor Mosquim
- Department of Operative Dentistry, Endodontics and Dental Materials, Bauru School of Dentistry, University of São Paulo, Bauru 17012-901, Brazil
| | | | - Guy Howard Carpenter
- Centre for Host-Microbiome Interactions, Faculty of Dental, Oral & Craniofacial Sciences, Guy’s Campus, King’s College London, London SE1 1UL, UK
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Hyvärinen E, Kashyap B, Kullaa AM. Oral Sources of Salivary Metabolites. Metabolites 2023; 13:metabo13040498. [PMID: 37110157 PMCID: PMC10145445 DOI: 10.3390/metabo13040498] [Citation(s) in RCA: 5] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/16/2023] [Revised: 03/10/2023] [Accepted: 03/28/2023] [Indexed: 03/31/2023] Open
Abstract
The oral cavity is very diverse, where saliva plays an important role in maintaining oral health. The metabolism of saliva has been used to investigate oral diseases as well as general diseases, mainly to detect diagnostic biomarkers. There are many sources of salivary metabolites in the mouth. The online English language search and PubMed databases were searched to retrieve relevant studies on oral salivary metabolites. The physiological balance of the mouth is influenced by many factors that are reflected in the salivary metabolite profile. Similarly, the dysbiosis of microbes can alter the salivary metabolite profile, which may express oral inflammation or oral diseases. This narrative review highlights the factors to be considered when examining saliva and its use as a diagnostic biofluid for different diseases. Salivary metabolites, mainly small molecular metabolites may enter the bloodstream and cause illness elsewhere in the body. The importance of salivary metabolites produced in the oral cavity as risk factors for general diseases and their possible relationship to the body’s function are also discussed.
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15
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Metabolic Profile of Whole Unstimulated Saliva in Patients with Sjögren’s Syndrome. Metabolites 2023; 13:metabo13030348. [PMID: 36984788 PMCID: PMC10054882 DOI: 10.3390/metabo13030348] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/05/2023] [Revised: 02/22/2023] [Accepted: 02/24/2023] [Indexed: 03/03/2023] Open
Abstract
(1) Background: Primary Sjögren’s Syndrome (pSS) is a multi-system autoimmune disease that involves the exocrine glands. Lymphocytes infiltrate the gland tissue, leading to anatomical modification and hypofunction. Even if the prognosis of pSS is favorable, quality of life is typically reduced due to the diverse manifestations of the disease. The aim of this study is to compare the salivary metabolomes of pSS with healthy controls (HCs). (2) Methods: Seven cases were selected from a cohort of pSS patients, and six age- and sex-matched HCs were recruited from a cohort of volunteers. Whole unstimulated saliva was collected for NMR analysis. Our metabolomic analysis focused on 360 ms total echo 1D 1H NMR CPMG spectra. Metabolites detected with CPMG NMR spectra were assigned through 2D NMR spectra (COSY, TOCSY, and HSQC). (3) Results: About 50 metabolites were detected and assigned. Unsupervised exploratory PCA returned partial clustering, and PLS-DA improved the separation between pSS and HCs, highlighting a pool of metabolites distinctly describing each group. (4) Conclusions: Despite the limited number of samples, the presented preliminary data are promising. PLS-DA indicated well-defined group separation, suggesting that the application of 1H-NMR metabolomics is suitable for the study of pSS.
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16
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Garwolińska D, Kot-Wasik A, Hewelt-Belka W. Pre-analytical aspects in metabolomics of human biofluids - sample collection, handling, transport, and storage. Mol Omics 2023; 19:95-104. [PMID: 36524542 DOI: 10.1039/d2mo00212d] [Citation(s) in RCA: 4] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/16/2022]
Abstract
Metabolomics is the field of omics research that offers valuable insights into the complex composition of biological samples. It has found wide application in clinical diagnostics, disease investigation, therapy prediction, monitoring of treatment efficiency, drug discovery, or in-depth analysis of sample composition. A suitable study design constitutes the fundamental requirements to ensure robust and reliable results from the study data. The study design process should include a careful selection of conditions for each experimental step, from sample collection to data analysis. The pre-analytical variability that can introduce bias to the subsequent analytical process, decrease the outcome reliability, and confuse the final results of the metabolomics research, should also be considered. Herein, we provide key information regarding the pre-analytical variables affecting the metabolomics studies of biological fluids that are the most desirable type of biological samples. Our work offers a practical review that can serve and guide metabolomics pre-analytical design. It indicates pre-analytical factors, which can introduce artificial data variation and should be identified and understood during experimental design (through literature overview or analytical experiments).
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Affiliation(s)
- Dorota Garwolińska
- Department of Analytical Chemistry, Faculty of Chemistry, Gdańsk University of Technology, Gabriela Narutowicza 11/12, 80-233 Gdańsk, Poland.
| | - Agata Kot-Wasik
- Department of Analytical Chemistry, Faculty of Chemistry, Gdańsk University of Technology, Gabriela Narutowicza 11/12, 80-233 Gdańsk, Poland.
| | - Weronika Hewelt-Belka
- Department of Analytical Chemistry, Faculty of Chemistry, Gdańsk University of Technology, Gabriela Narutowicza 11/12, 80-233 Gdańsk, Poland.
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Huang Z, Yang X, Huang Y, Tang Z, Chen Y, Liu H, Huang M, Qing L, Li L, Wang Q, Jie Z, Jin X, Jia B. Saliva - a new opportunity for fluid biopsy. Clin Chem Lab Med 2023; 61:4-32. [PMID: 36285724 DOI: 10.1515/cclm-2022-0793] [Citation(s) in RCA: 26] [Impact Index Per Article: 13.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/11/2022] [Accepted: 09/29/2022] [Indexed: 12/15/2022]
Abstract
Saliva is a complex biological fluid with a variety of biomolecules, such as DNA, RNA, proteins, metabolites and microbiota, which can be used for the screening and diagnosis of many diseases. In addition, saliva has the characteristics of simple collection, non-invasive and convenient storage, which gives it the potential to replace blood as a new main body of fluid biopsy, and it is an excellent biological diagnostic fluid. This review integrates recent studies and summarizes the research contents of salivaomics and the research progress of saliva in early diagnosis of oral and systemic diseases. This review aims to explore the value and prospect of saliva diagnosis in clinical application.
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Affiliation(s)
- Zhijie Huang
- Department of Oral Surgery, Stomatological Hospital, Southern Medical University, Guangzhou, P.R. China
| | - Xiaoxia Yang
- Department of Endodontics, Stomatological Hospital, Southern Medical University, Guangzhou, P.R. China
| | - Yisheng Huang
- Department of Oral Surgery, Stomatological Hospital, Southern Medical University, Guangzhou, P.R. China
| | - Zhengming Tang
- Department of Oral Surgery, Stomatological Hospital, Southern Medical University, Guangzhou, P.R. China
| | - Yuanxin Chen
- Department of Oral Surgery, Stomatological Hospital, Southern Medical University, Guangzhou, P.R. China
| | - Hongyu Liu
- Department of Oral Surgery, Stomatological Hospital, Southern Medical University, Guangzhou, P.R. China
| | - Mingshu Huang
- Department of Oral Surgery, Stomatological Hospital, Southern Medical University, Guangzhou, P.R. China
| | - Ling Qing
- Department of Oral Surgery, Stomatological Hospital, Southern Medical University, Guangzhou, P.R. China
| | - Li Li
- Department of Oral Surgery, Stomatological Hospital, Southern Medical University, Guangzhou, P.R. China
| | - Qin Wang
- Department of Oral Surgery, Stomatological Hospital, Southern Medical University, Guangzhou, P.R. China
| | - Zhuye Jie
- BGI Genomics, BGI-Shenzhen, Shenzhen, P.R. China
- Shenzhen Key Laboratory of Human Commensal Microorganisms and Health Research, BGI-Shenzhen, Shenzhen, P.R. China
- Laboratory of Genomics and Molecular Biomedicine, Department of Biology, University of Copenhagen, Copenhagen, Denmark
| | - Xin Jin
- BGI Genomics, BGI-Shenzhen, Shenzhen, P.R. China
- School of Medicine, South China University of Technology, Guangzhou, P.R. China
| | - Bo Jia
- Department of Oral Surgery, Stomatological Hospital, Southern Medical University, Guangzhou, P.R. China
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18
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Qualitative and Quantitative Mass Spectrometry in Salivary Metabolomics and Proteomics. Metabolites 2023; 13:metabo13020155. [PMID: 36837774 PMCID: PMC9964739 DOI: 10.3390/metabo13020155] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/02/2022] [Revised: 01/11/2023] [Accepted: 01/17/2023] [Indexed: 01/22/2023] Open
Abstract
The metabolomics and proteomics analysis of saliva, an excellent biofluid that is a rich source of biological compounds, allows for the safe and frequent screening of drugs, their metabolites, and molecular biomarkers of various diseases. One of the most frequently used analytical methods in saliva analysis is liquid chromatography coupled with mass spectrometry (LC-MS) and tandem mass spectrometry. The low ionisation efficiency of some compounds and a complex matrix makes their identification by MS difficult. Furthermore, quantitative analysis by LC-MS frequently cannot be performed without isotopically labelled standards, which usually have to be specially synthesised. This review presented reports on qualitative and quantitative approaches in salivary metabolomics and proteomics. The purpose of this manuscript was to present the challenges, advances, and future prospects of mass spectrometry, both in the analysis of salivary metabolites and proteins. The presented review should appeal to those interested in the recent advances and trends in qualitative and quantitative mass spectrometry in salivary metabolomics and proteomics, which may facilitate a diagnostic accuracy, the evaluation of treatment efficacy, the early diagnosis of disease, and a forensic investigation of some unapproved drugs for any medical or dietary administration.
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Sugimoto M, Aizawa Y, Tomita A. Data Processing and Analysis in Liquid Chromatography-Mass Spectrometry-Based Targeted Metabolomics. Methods Mol Biol 2023; 2571:241-255. [PMID: 36152165 DOI: 10.1007/978-1-0716-2699-3_21] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 06/16/2023]
Abstract
Mass spectrometry (MS)-based metabolomics provides high-dimensional datasets; that is, the data include various metabolite features. Data analysis begins by converting the raw data obtained from the MS to produce a data matrix (metabolite × concentrations). This is followed by several steps, such as peak integration, alignment of multiple data, metabolite identification, and calculation of metabolite concentrations. Each step yields the analytical results and the accompanying information used for the quality assessment of the anterior steps. Thus, the measurement quality can be analyzed through data processing. Here, we introduce a typical data processing procedure and describe a method to utilize the intermediate data as quality control. Subsequently, commonly used data analysis methods for metabolomics data, such as statistical analyses, are also introduced.
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Affiliation(s)
- Masahiro Sugimoto
- Institute of Medical Science, Tokyo Medical University, Tokyo, Japan.
- Institute for Advanced Biosciences, Yamagata, Japan.
| | - Yumi Aizawa
- Institute of Medical Science, Tokyo Medical University, Tokyo, Japan
| | - Atsumi Tomita
- Institute of Medical Science, Tokyo Medical University, Tokyo, Japan
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20
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Grootveld M, Page G, Bhogadia M, Hunwin K, Edgar M. Updates and Original Case Studies Focused on the NMR-Linked Metabolomics Analysis of Human Oral Fluids Part III: Implementations for the Diagnosis of Non-Cancerous Disorders, Both Oral and Systemic. Metabolites 2023; 13:metabo13010066. [PMID: 36676991 PMCID: PMC9864626 DOI: 10.3390/metabo13010066] [Citation(s) in RCA: 4] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/20/2022] [Revised: 12/27/2022] [Accepted: 12/28/2022] [Indexed: 01/03/2023] Open
Abstract
This communication represents Part III of our series of reports based on the applications of human saliva as a useful and conveniently collectable medium for the discovery, identification and monitoring of biomarkers, which are of some merit for the diagnosis of human diseases. Such biomarkers, or others reflecting the dysfunction of specific disease-associated metabolic pathways, may also be employed for the prognostic pathological tracking of these diseases. Part I of this series set the experimental and logistical groundwork for this report, and the preceding paper, Part II, featured the applications of newly developed metabolomics technologies to the diagnosis and severity grading of human cancer conditions, both oral and systemic. Clearly, there are many benefits, both scientific and economic, associated with the donation of human saliva samples (usually as whole mouth saliva) from humans consenting to and participating in investigations focused on the discovery of biomolecular markers of diseases. These include usually non-invasive collection protocols, relatively low cost when compared against blood sample collection, and no requirement for clinical supervision during collection episodes. This paper is centred on the employment and value of 'state-of-the-art' metabolomics technologies to the diagnosis and prognosis of a wide range of non-cancerous human diseases. Firstly, these include common oral diseases such as periodontal diseases (from type 1 (gingivitis) to type 4 (advanced periodontitis)), and dental caries. Secondly, a wide range of extra-oral (systemic) conditions are covered, most notably diabetes types 1 and 2, cardiovascular and neurological diseases, and Sjögren's syndrome, along with a series of viral infections, e.g., pharyngitis, influenza, HIV and COVID-19. Since the authors' major research interests lie in the area of the principles and applications of NMR-linked metabolomics techniques, many, but not all, of the studies reviewed were conducted using these technologies, with special attention being given to recommended protocols for their operation and management, for example, satisfactory experimental model designs; sample collection and laboratory processing techniques; the selection of sample-specific NMR pulse sequences for saliva analysis; and strategies available for the confirmation of resonance assignments for both endogenous and exogenous molecules in this biofluid. This article also features an original case study, which is focussed on the use of NMR-based salivary metabolomics techniques to provide some key biomarkers for the diagnosis of pharyngitis, and an example of how to 'police' such studies and to recognise participants who perceive that they actually have this disorder but do not from their metabolic profiles and multivariate analysis pattern-based clusterings. The biochemical and clinical significance of these multidimensional metabolomics investigations are discussed in detail.
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Foratori-Junior GA, Guennec AL, Fidalgo TKDS, Cleaver L, Buzalaf MAR, Carpenter GH, Sales-Peres SHDC. Metabolomic Profiles Associated with Obesity and Periodontitis during Pregnancy: Cross-Sectional Study with Proton Nuclear Magnetic Resonance ( 1H-NMR)-Based Analysis. Metabolites 2022; 12:metabo12111029. [PMID: 36355112 PMCID: PMC9694155 DOI: 10.3390/metabo12111029] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/29/2022] [Revised: 10/18/2022] [Accepted: 10/24/2022] [Indexed: 12/27/2022] Open
Abstract
This study aimed to elucidate the metabolomic signature associated with obesity and periodontitis during pregnancy in plasma and saliva biofluids. Ninety-eight pregnant women were divided into: with obesity and periodontitis (OP = 20), with obesity but without periodontitis (OWP = 27), with normal BMI but with periodontitis (NP = 21), with normal BMI and without periodontitis (NWP = 30). Saliva and plasma were analyzed by 1H-NMR for metabolites identification. Partial Least Squares-Discriminant Analysis (PLS-DA), Sparse PLS-DA (sPLS-DA), and Variable Importance of Projection (VIP) were performed. ANOVA and Pearson’s correlation were applied (p < 0.05). Plasmatic analysis indicated the levels of glucose (p = 0.041) and phenylalanine (p = 0.015) were positively correlated with periodontal parameters and BMI, respectively. In saliva, periodontitis was mainly associated with high levels of acetic acid (p = 0.024), isovaleric acid, butyric acid, leucine, valine, isoleucine, and propionic acid (p < 0.001). High salivary concentrations of glycine (p = 0.015), succinic acid (p = 0.015), and lactate (p = 0.026) were associated with obesity. Saliva demonstrated a more elucidative difference than plasma, indicating the glucose-alanine cycle, alanine metabolism, valine, leucine and isoleucine degradation, glutamate metabolism, and Warburg effect as the main metabolic pathways.
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Affiliation(s)
- Gerson Aparecido Foratori-Junior
- Department of Pediatric Dentistry, Orthodontics and Public Health, Bauru School of Dentistry, University of São Paulo, Bauru 17012-901, Brazil
- Centre for Host-Microbiome Interactions, Faculty of Dental, Oral & Craniofacial Sciences, Guy’s Campus, King’s College London, London SE1 1UL, UK
- Correspondence: (G.A.F.-J.); (S.H.d.C.S.-P.)
| | - Adrien Le Guennec
- Nuclear Magnetic Resonance Facility, Guy’s Campus, King’s College London, London SE1 1UL, UK
| | - Tatiana Kelly da Silva Fidalgo
- Department of Preventive and Community Dentistry, School of Dentistry, Rio de Janeiro State University, Rio de Janeiro 20551-030, Brazil
| | - Leanne Cleaver
- Centre for Host-Microbiome Interactions, Faculty of Dental, Oral & Craniofacial Sciences, Guy’s Campus, King’s College London, London SE1 1UL, UK
| | | | - Guy Howard Carpenter
- Centre for Host-Microbiome Interactions, Faculty of Dental, Oral & Craniofacial Sciences, Guy’s Campus, King’s College London, London SE1 1UL, UK
| | - Silvia Helena de Carvalho Sales-Peres
- Department of Pediatric Dentistry, Orthodontics and Public Health, Bauru School of Dentistry, University of São Paulo, Bauru 17012-901, Brazil
- Correspondence: (G.A.F.-J.); (S.H.d.C.S.-P.)
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Updates and Original Case Studies Focused on the NMR-Linked Metabolomics Analysis of Human Oral Fluids Part II: Applications to the Diagnosis and Prognostic Monitoring of Oral and Systemic Cancers. Metabolites 2022; 12:metabo12090778. [PMID: 36144183 PMCID: PMC9505390 DOI: 10.3390/metabo12090778] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/30/2022] [Revised: 08/12/2022] [Accepted: 08/15/2022] [Indexed: 11/24/2022] Open
Abstract
Human saliva offers many advantages over other biofluids regarding its use and value as a bioanalytical medium for the identification and prognostic monitoring of human diseases, mainly because its collection is largely non-invasive, is relatively cheap, and does not require any major clinical supervision, nor supervisory input. Indeed, participants donating this biofluid for such purposes, including the identification, validation and quantification of surrogate biomarkers, may easily self-collect such samples in their homes following the provision of full collection details to them by researchers. In this report, the authors have focused on the applications of metabolomics technologies to the diagnosis and progressive severity monitoring of human cancer conditions, firstly oral cancers (e.g., oral cavity squamous cell carcinoma), and secondly extra-oral (systemic) cancers such as lung, breast and prostate cancers. For each publication reviewed, the authors provide a detailed evaluation and critical appraisal of the experimental design, sample size, ease of sample collection (usually but not exclusively as whole mouth saliva (WMS)), their transport, length of storage and preparation for analysis. Moreover, recommended protocols for the optimisation of NMR pulse sequences for analysis, along with the application of methods and techniques for verifying and resonance assignments and validating the quantification of biomolecules responsible, are critically considered. In view of the authors’ specialisms and research interests, the majority of these investigations were conducted using NMR-based metabolomics techniques. The extension of these studies to determinations of metabolic pathways which have been pathologically disturbed in these diseases is also assessed here and reviewed. Where available, data for the monitoring of patients’ responses to chemotherapeutic treatments, and in one case, radiotherapy, are also evaluated herein. Additionally, a novel case study featured evaluates the molecular nature, levels and diagnostic potential of 1H NMR-detectable salivary ‘acute-phase’ glycoprotein carbohydrate side chains, and/or their monomeric saccharide derivatives, as biomarkers for cancer and inflammatory conditions.
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Panneerselvam K, Ishikawa S, Krishnan R, Sugimoto M. Salivary Metabolomics for Oral Cancer Detection: A Narrative Review. Metabolites 2022; 12:metabo12050436. [PMID: 35629940 PMCID: PMC9144467 DOI: 10.3390/metabo12050436] [Citation(s) in RCA: 12] [Impact Index Per Article: 4.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/30/2022] [Revised: 05/07/2022] [Accepted: 05/11/2022] [Indexed: 12/24/2022] Open
Abstract
The development of low- or non-invasive screening tests for cancer is crucial for early detection. Saliva is an ideal biofluid containing informative components for monitoring oral and systemic diseases. Metabolomics has frequently been used to identify and quantify numerous metabolites in saliva samples, serving as novel biomarkers associated with various conditions, including cancers. This review summarizes the recent applications of salivary metabolomics in biomarker discovery in oral cancers. We discussed the prevalence, epidemiologic characteristics, and risk factors of oral cancers, as well as the currently available screening programs, in India and Japan. These data imply that the development of biomarkers by itself is inadequate in cancer detection. The use of current diagnostic methods and new technologies is necessary for efficient salivary metabolomics analysis. We also discuss the gap between biomarker discovery and nationwide screening for the early detection of oral cancer and its prevention.
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Affiliation(s)
- Karthika Panneerselvam
- Department of Oral Pathology and Microbiology, Karpaga Vinayaga Institute of Dental Sciences, GST Road, Chinna Kolambakkam, Palayanoor PO, Madurantagam Taluk, Kancheepuram 603308, Tamil Nadu, India;
| | - Shigeo Ishikawa
- Department of Dentistry, Oral and Maxillofacial Plastic and Reconstructive Surgery, Faculty of Medicine, Yamagata University, Yamagata 990-9585, Japan;
| | - Rajkumar Krishnan
- Department of Oral Pathology, SRM Dental College, Bharathi Salai, Ramapuram, Chennai 600089, Tamil Nadu, India;
| | - Masahiro Sugimoto
- Institute of Medical Research, Tokyo Medical University, Tokyo 160-0022, Japan
- Institute for Advanced Biosciences, Keio University, Yamagata 997-0811, Japan
- Correspondence: ; Tel.: +81-235-29-0528
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Ejaz S, Ejaz S, Shahid R, Noor T, Shabbir S, Imran M. Chitosan-curcumin complexation to develop functionalized nanosystems with enhanced antimicrobial activity against hetero-resistant gastric pathogen. Int J Biol Macromol 2022; 204:540-554. [PMID: 35157901 DOI: 10.1016/j.ijbiomac.2022.02.039] [Citation(s) in RCA: 15] [Impact Index Per Article: 5.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/11/2021] [Revised: 02/01/2022] [Accepted: 02/08/2022] [Indexed: 12/13/2022]
Abstract
With the apparent stagnation in the antibiotic discovery and the propagation of multidrug resistance, Helicobacter pylori associated gastric infections are hard to eradicate. In pursuance of alternative medicines, in this study, covalent modification of chitosan (CS) polymer with curcumin (Cur) was accomplished. Proton Nuclear Magnetic Resonance and Fourier Transform Infrared spectroscopy elucidated the covalent interaction between Cur and CS with characteristic peak of imine functional group (C=N). Scanning Electron Microscopy provided visual proof for surface topology, while size and zeta potential values further affirmed the development of curcumin functionalized chitosan nanosystems (Cur-FCNS). The complexation efficiency of CS with Cur was found as 70 ± 3% at an optimal ratio of 5:1 for CS and Cur, respectively. Cur-FCNS developed with ionic gelation and ultrasonication method demonstrated synergistic anti-H. pylori activity in growth-kinetics and anti-biofilm assays, which was superior to free Cur and even chitosan nanosystems. Under simulated gastric conditions, Cur-FCNS revealed cumulative-release of only 16 ± 0.8% till 40 h, which indicated its improved stability to interact with H. pylori. In silico findings affirmed high binding affinity of Cur-FCNS with multiple bacterial virulence factors. Thus, our results affirmed the exceptional potential of Cur-FCNS as next-generation alternative-medicine to treat resistant H. pylori.
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Affiliation(s)
- Sadaf Ejaz
- Department of Biosciences, Faculty of Sciences, COMSATS University Islamabad (CUI), Park Road, Islamabad, Pakistan
| | - Saima Ejaz
- Research Centre for Modelling and Simulation (RCMS), National University of Science and Technology (NUST), Islamabad, Pakistan
| | - Ramla Shahid
- Department of Biosciences, Faculty of Sciences, COMSATS University Islamabad (CUI), Park Road, Islamabad, Pakistan
| | - Tayyaba Noor
- School of Chemical and Materials Engineering (SCME), National University of Science and Technology (NUST), Islamabad, Pakistan
| | - Saima Shabbir
- Department of Materials Science and Engineering, Institute of Space Technology (IST), Islamabad 44000, Pakistan
| | - Muhammad Imran
- Department of Biosciences, Faculty of Sciences, COMSATS University Islamabad (CUI), Park Road, Islamabad, Pakistan.
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25
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Quartieri E, Casali E, Ferrari E, Ghezzi B, Gallo M, Spisni A, Meleti M, Pertinhez TA. Sample optimization for saliva 1H-NMR metabolic profiling. Anal Biochem 2022; 640:114412. [PMID: 34656613 DOI: 10.1016/j.ab.2021.114412] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/29/2020] [Revised: 09/06/2021] [Accepted: 10/08/2021] [Indexed: 11/01/2022]
Abstract
Nuclear Magnetic Resonance (NMR) based metabolomic analysis of whole saliva has provided potential diagnostic biomarkers for numerous human diseases contributing to a better understanding of their mechanisms. However, a comprehensive interpretation of the significance of metabolites in whole, parotid, and submandibular/sublingual saliva subtypes is still missing. Precision and reproducibility of sample preparation is an essential step. Here, we present a simple and efficient protocol for saliva 1H-NMR metabolic profiling. This procedure has been specifically designed and optimized for the identification and quantification of low concentration metabolites (as low as 1.1 μM) and is suitable for all the saliva subtypes.
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Affiliation(s)
- Eleonora Quartieri
- Department of Medicine and Surgery, University of Parma, Parma, Italy; Transfusion Medicine Unit, Azienda USL-IRCCS di Reggio Emilia, Reggio Emilia, Italy
| | - Emanuela Casali
- Department of Medicine and Surgery, University of Parma, Parma, Italy
| | - Elena Ferrari
- Department of Medicine and Surgery, University of Parma, Parma, Italy.
| | - Benedetta Ghezzi
- Department of Medicine and Surgery, University of Parma, Parma, Italy; Centro Universitario di Odontoiatria, University of Parma, Parma, Italy
| | - Mariana Gallo
- Department of Medicine and Surgery, University of Parma, Parma, Italy
| | - Alberto Spisni
- Department of Medicine and Surgery, University of Parma, Parma, Italy
| | - Marco Meleti
- Department of Medicine and Surgery, University of Parma, Parma, Italy; Centro Universitario di Odontoiatria, University of Parma, Parma, Italy
| | - Thelma A Pertinhez
- Department of Medicine and Surgery, University of Parma, Parma, Italy; Transfusion Medicine Unit, Azienda USL-IRCCS di Reggio Emilia, Reggio Emilia, Italy
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Predicting Satiety from the Analysis of Human Saliva Using Mid-Infrared Spectroscopy Combined with Chemometrics. Foods 2022; 11:foods11050711. [PMID: 35267343 PMCID: PMC8909147 DOI: 10.3390/foods11050711] [Citation(s) in RCA: 3] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/10/2022] [Revised: 02/24/2022] [Accepted: 02/24/2022] [Indexed: 02/04/2023] Open
Abstract
The aim of this study was to evaluate the ability of mid-infrared (MIR) spectroscopy combined with chemometrics to analyze unstimulated saliva as a method to predict satiety in healthy participants. This study also evaluated features in saliva that were related to individual perceptions of human–food interactions. The coefficient of determination (R2) and standard error in cross validation (SECV) for the prediction of satiety in all saliva samples were 0.62 and 225.7 satiety area under the curve (AUC), respectively. A correlation between saliva and satiety was found, however, the quantitative prediction of satiety using unstimulated saliva was not robust. Differences in the MIR spectra of saliva between low and high satiety groups, were observed in the following frequency ratios: 1542/2060 cm−1 (total protein), 1637/3097 cm−1 (α-amino acids), and 1637/616 (chlorides) cm−1. In addition, good to excellent models were obtained for the prediction of satiety groups defined as low or high satiety participants (R2 0.92 and SECV 0.10), demonstrating that this method could be used to identify low or high satiety perception types and to select participants for appetite studies. Although quantitative PLS calibration models were not achieved, a qualitative model for the prediction of low and high satiety perception types was obtained using PLS-DA. Furthermore, this study showed that it might be possible to evaluate human/food interactions using MIR spectroscopy as a rapid and cost-effective tool.
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Updates and Original Case Studies Focused on the NMR-Linked Metabolomics Analysis of Human Oral Fluids Part I: Emerging Platforms and Perspectives. APPLIED SCIENCES-BASEL 2022. [DOI: 10.3390/app12031235] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 02/06/2023]
Abstract
1H NMR-based metabolomics analysis of human saliva, other oral fluids, and/or tissue biopsies serves as a valuable technique for the exploration of metabolic processes, and when associated with ’state-of-the-art’ multivariate (MV) statistical analysis strategies, provides a powerful means of examining the identification of characteristic metabolite patterns, which may serve to differentiate between patients with oral health conditions (e.g., periodontitis, dental caries, and oral cancers) and age-matched heathy controls. This approach may also be employed to explore such discriminatory signatures in the salivary 1H NMR profiles of patients with systemic diseases, and to date, these have included diabetes, Sjörgen’s syndrome, cancers, neurological conditions such as Alzheimer’s disease, and viral infections. However, such investigations are complicated in view of quite a large number of serious inconsistencies between the different studies performed by independent research groups globally; these include differing protocols and routes for saliva sample collection (e.g., stimulated versus unstimulated samples), their timings (particularly the oral activity abstention period involved, which may range from one to 12 h or more), and methods for sample transport, storage, and preparation for NMR analysis, not to mention a very wide variety of demographic variables that may influence salivary metabolite concentrations, notably the age, gender, ethnic origin, salivary flow-rate, lifestyles, diets, and smoking status of participant donors, together with their exposure to any other possible convoluting environmental factors. In view of the explosive increase in reported salivary metabolomics investigations, in this update, we critically review a wide range of critical considerations for the successful performance of such experiments. These include the nature, composite sources, and biomolecular status of human saliva samples; the merits of these samples as media for the screening of disease biomarkers, notably their facile, unsupervised collection; and the different classes of such metabolomics investigations possible. Also encompassed is an account of the history of NMR-based salivary metabolomics; our recommended regimens for the collection, transport, and storage of saliva samples, along with their preparation for NMR analysis; frequently employed pulse sequences for the NMR analysis of these samples; the supreme resonance assignment benefits offered by homo- and heteronuclear two-dimensional NMR techniques; deliberations regarding salivary biomolecule quantification approaches employed for such studies, including the preprocessing and bucketing of multianalyte salivary NMR spectra, and the normalization, transformation, and scaling of datasets therefrom; salivary phenotype analysis, featuring the segregation of a range of different metabolites into ‘pools’ grouped according to their potential physiological sources; and lastly, future prospects afforded by the applications of LF benchtop NMR spectrometers for direct evaluations of the oral or systemic health status of patients at clinical ‘point-of-contact’ sites, e.g., dental surgeries. This commentary is then concluded with appropriate recommendations for the conduct of future salivary metabolomics studies. Also included are two original case studies featuring investigations of (1) the 1H NMR resonance line-widths of selected biomolecules and their possible dependence on biomacromolecular binding equilibria, and (2) the combined univariate (UV) and MV analysis of saliva specimens collected from a large group of healthy control participants in order to potentially delineate the possible origins of biomolecules therein, particularly host- versus oral microbiome-derived sources. In a follow-up publication, Part II of this series, we conduct censorious reviews of reported observations acquired from a diversity of salivary metabolomics investigations performed to evaluate both localized oral and non-oral diseases. Perplexing problems encountered with these again include those arising from sample collection and preparation protocols, along with 1H NMR spectral misassignments.
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Vilca-Melendez S, Uthaug MV, Griffin JL. 1H Nuclear Magnetic Resonance: A Future Approach to the Metabolic Profiling of Psychedelics in Human Biofluids? Front Psychiatry 2021; 12:742856. [PMID: 34966300 PMCID: PMC8710695 DOI: 10.3389/fpsyt.2021.742856] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 07/19/2021] [Accepted: 11/18/2021] [Indexed: 11/25/2022] Open
Abstract
While psychedelics may have therapeutic potential for treating mental health disorders such as depression, further research is needed to better understand their biological effects and mechanisms of action when considering the development of future novel therapy approaches. Psychedelic research could potentially benefit from the integration of metabonomics by proton nuclear magnetic resonance (1H NMR) spectroscopy which is an analytical chemistry-based approach that can measure the breakdown of drugs into their metabolites and their metabolic consequences from various biofluids. We have performed a systematic review with the primary aim of exploring published literature where 1H NMR analysed psychedelic substances including psilocin, lysergic acid diethylamide (LSD), LSD derivatives, N,N-dimethyltryptamine (DMT), 5-methoxy-N,N-dimethyltryptamine (5-MeO-DMT) and bufotenin. The second aim was to assess the benefits and limitations of 1H NMR spectroscopy-based metabolomics as a tool in psychedelic research and the final aim was to explore potential future directions. We found that the most current use of 1H NMR in psychedelic research has been for the structural elucidation and analytical characterisation of psychedelic molecules and that no papers used 1H NMR in the metabolic profiling of biofluids, thus exposing a current research gap and the underuse of 1H NMR. The efficacy of 1H NMR spectroscopy was also compared to mass spectrometry, where both metabonomics techniques have previously shown to be appropriate for biofluid analysis in other applications. Additionally, potential future directions for psychedelic research were identified as real-time NMR, in vivo 1H nuclear magnetic resonance spectroscopy (MRS) and 1H NMR studies of the gut microbiome. Further psychedelic studies need to be conducted that incorporate the use of 1H NMR spectroscopy in the analysis of metabolites both in the peripheral biofluids and in vivo to determine whether it will be an effective future approach for clinical and naturalistic research.
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Affiliation(s)
- Sylvana Vilca-Melendez
- Department of Brain Sciences, Faculty of Medicine, Imperial College London, London, United Kingdom
| | - Malin V. Uthaug
- The Centre for Psychedelic Research, Department of Brain Sciences, Faculty of Medicine, Imperial College London, London, United Kingdom
- Department of Neuropsychology and Psychopharmacology, Faculty of Psychology and Neuroscience, Maastricht University, Maastricht, Netherlands
| | - Julian L. Griffin
- Department of Metabolism, Digestion and Reproduction, Faculty of Medicine, Imperial College London, London, United Kingdom
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Duś-Ilnicka I, Krala E, Cholewińska P, Radwan-Oczko M. The Use of Saliva as a Biosample in the Light of COVID-19. Diagnostics (Basel) 2021; 11:1769. [PMID: 34679467 PMCID: PMC8534561 DOI: 10.3390/diagnostics11101769] [Citation(s) in RCA: 10] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/10/2021] [Revised: 09/18/2021] [Accepted: 09/22/2021] [Indexed: 12/13/2022] Open
Abstract
Saliva is easy to collect and a biofluid that is readily available without the need for special equipment for its collection. The collection process, which is non-invasive and inexpensive, leads to obtaining a biomaterial that can serve as a source of information for molecular diagnostics of diseases in general medicine, genetics and dentistry. Unfortunately, many of the salivary methodologies are lacking important parameters to provide for not only the safety of the operator, but also the quality and reproducibility of the research. Since the COVID-19 pandemic, salivary diagnostics demonstrate a great potential for research of SARS-CoV 2. In this review, good practice for unstimulated saliva collection and patient preparation was provided, based on the latest literature and available guidelines. Schemes for saliva collection procedures were presented following an extended literature search. Descriptions of salivary probes/cups, techniques of saliva collection, and the use of specific buffering solutions for the stability of collected samples for SARS-CoV-2 detection were also evaluated.
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Affiliation(s)
- Irena Duś-Ilnicka
- Oral Pathology Department, Faculty of Dentistry, Wroclaw Medical University, ul. Krakowska 26, 50-425 Wroclaw, Poland; (E.K.); (M.R.-O.)
| | - Elżbieta Krala
- Oral Pathology Department, Faculty of Dentistry, Wroclaw Medical University, ul. Krakowska 26, 50-425 Wroclaw, Poland; (E.K.); (M.R.-O.)
| | - Paulina Cholewińska
- Institute of Animal Breeding, Faculty of Biology and Animal Breeding, Wroclaw University of Enviromental and Life Sciences, ul. Chełmońskiego 38C, 51-630 Wroclaw, Poland;
| | - Małgorzata Radwan-Oczko
- Oral Pathology Department, Faculty of Dentistry, Wroclaw Medical University, ul. Krakowska 26, 50-425 Wroclaw, Poland; (E.K.); (M.R.-O.)
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Salivary Metabolomics for Diagnosis and Monitoring Diseases: Challenges and Possibilities. Metabolites 2021; 11:metabo11090587. [PMID: 34564402 PMCID: PMC8469343 DOI: 10.3390/metabo11090587] [Citation(s) in RCA: 44] [Impact Index Per Article: 11.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/08/2021] [Revised: 08/27/2021] [Accepted: 08/28/2021] [Indexed: 12/21/2022] Open
Abstract
Saliva is a useful biological fluid and a valuable source of biological information. Saliva contains many of the same components that can be found in blood or serum, but the components of interest tend to be at a lower concentration in saliva, and their analysis demands more sensitive techniques. Metabolomics is starting to emerge as a viable method for assessing the salivary metabolites which are generated by the biochemical processes in elucidating the pathways underlying different oral and systemic diseases. In oral diseases, salivary metabolomics has concentrated on periodontitis and oral cancer. Salivary metabolites of systemic diseases have been investigated mostly in the early diagnosis of different cancer, but also neurodegenerative diseases. This mini-review article aims to highlight the challenges and possibilities of salivary metabolomics from a clinical viewpoint. Furthermore, applications of the salivary metabolic profile in diagnosis and prognosis, monitoring the treatment success, and planning of personalized treatment of oral and systemic diseases are discussed.
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31
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Wren S, Collins L, Hughes LP, Jones I. Measuring the rate of in-vitro drug release from polymeric nanoparticles by 19F solution state NMR spectroscopy. J Pharm Sci 2021; 110:3546-3549. [PMID: 34400184 DOI: 10.1016/j.xphs.2021.08.011] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/24/2021] [Revised: 08/10/2021] [Accepted: 08/10/2021] [Indexed: 11/24/2022]
Abstract
We report what we believe is the first use of 19F NMR spectroscopy to directly measure in-vitro release (IVR) from polymeric nanoparticles (PNPs). Using 19F NMR we selectively measured IVR of AZD2811 from PNPs. Due to rapid nuclear relaxation in solid-like environments only AZD2811 in solution is detected, and physical separation from the PNPs isn't required. The NMR approach and ultra-centrifugation/UHPLC were shown to be equivalent. The selectivity of 19F NMR means it is readily applied to complex IVR media such as recombinant human serum albumin (rHSA).
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Affiliation(s)
- Stephen Wren
- New Modalities and Parenterals Development, Pharmaceutical Technology & Development, AstraZeneca, Macclesfield, UK.
| | - Laura Collins
- New Modalities and Parenterals Development, Pharmaceutical Technology & Development, AstraZeneca, Macclesfield, UK
| | - Leslie P Hughes
- Oral Product Development, Pharmaceutical Technology & Development, AstraZeneca, Macclesfield, UK
| | - Ian Jones
- New Modalities and Parenterals Development, Pharmaceutical Technology & Development, AstraZeneca, Macclesfield, UK
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32
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Havsed K, Stensson M, Jansson H, Carda-Diéguez M, Pedersen A, Neilands J, Svensäter G, Mira A. Bacterial Composition and Metabolomics of Dental Plaque From Adolescents. Front Cell Infect Microbiol 2021; 11:716493. [PMID: 34395316 PMCID: PMC8362896 DOI: 10.3389/fcimb.2021.716493] [Citation(s) in RCA: 38] [Impact Index Per Article: 9.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/28/2021] [Accepted: 07/12/2021] [Indexed: 01/04/2023] Open
Abstract
Supragingival dental plaque samples were collected from 40 Swedish adolescents, including 20 with caries lesions (CAR) and 20 caries-free (CF). Fresh plaque samples were subjected to an ex vivo acid tolerance (AT) test where the proportion of bacteria resistant to an acid shock was evaluated through confocal microscopy and live/dead staining, and the metabolites produced were quantified by 1H Nuclear Magnetic Resonance (1H NMR). In addition, DNA was extracted and the 16S rRNA gene was sequenced by Illumina sequencing, in order to characterize bacterial composition in the same samples. There were no significant differences in AT scores between CAR and CF individuals. However, 7 out of the 10 individuals with highest AT scores belonged to the CAR group. Regarding bacterial composition, Abiotrophia, Prevotella and Veillonella were found at significantly higher levels in CAR individuals (p=0.0085, 0.026 and 0.04 respectively) and Rothia and Corynebacterium at significantly higher levels in CF individuals (p=0.026 and 0.003). The caries pathogen Streptococcus mutans was found at low frequencies and was absent in 60% of CAR individuals. Random-forest predictive models indicate that at least 4 bacterial species or 9 genera are needed to distinguish CAR from CF adolescents. The metabolomic profile obtained by NMR showed a significant clustering of organic acids with specific bacteria in CAR and/or high AT individuals, being Scardovia wiggsiae the species with strongest associations. A significant clustering of ethanol and isopropanol with health-associated bacteria such as Rothia or Corynebacterium was also found. Accordingly, several relationships involving these compounds like the Ethanol : Lactate or Succinate : Lactate ratios were significantly associated to acid tolerance and could be of predictive value for caries risk. We therefore propose that future caries risk studies would benefit from considering not only the use of multiple organisms as potential microbial biomarkers, but also their functional adaptation and metabolic output.
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Affiliation(s)
- Kristian Havsed
- Department of Pediatric Dentistry, Institute for Postgraduate Dental Education, Jönköping, Sweden.,Centre for Oral Health, School of Health and Welfare, Jönköping University, Jönköping, Sweden.,Section for Oral Biology and Pathology, Faculty of Odontology, Malmö University, Malmö, Sweden
| | - Malin Stensson
- Centre for Oral Health, School of Health and Welfare, Jönköping University, Jönköping, Sweden.,Folktandvården Skåne, The Swedish Dental Service of Skåne, Lund, Sweden
| | - Henrik Jansson
- Centre for Oral Health, School of Health and Welfare, Jönköping University, Jönköping, Sweden.,Folktandvården Skåne, The Swedish Dental Service of Skåne, Lund, Sweden.,Department of Periodontology, Faculty of Odontology, Malmö University, Malmö, Sweden
| | - Miguel Carda-Diéguez
- Department of Health & Genomics, Foundation for the Promotion of Health and Biomedical Research (FISABIO) Foundation, Valencia, Spain
| | - Anders Pedersen
- Swedish NMR Centre, The University of Gothenburg, Gothenburg, Sweden
| | - Jessica Neilands
- Section for Oral Biology and Pathology, Faculty of Odontology, Malmö University, Malmö, Sweden.,Biofilms - Research Center for Biointerfaces, Malmö University, Malmö, Sweden
| | - Gunnel Svensäter
- Section for Oral Biology and Pathology, Faculty of Odontology, Malmö University, Malmö, Sweden.,Biofilms - Research Center for Biointerfaces, Malmö University, Malmö, Sweden
| | - Alex Mira
- Centre for Oral Health, School of Health and Welfare, Jönköping University, Jönköping, Sweden.,Department of Health & Genomics, Foundation for the Promotion of Health and Biomedical Research (FISABIO) Foundation, Valencia, Spain
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33
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Rovera A, Hector M, Anderson P. Parotid saliva 1 H-NMR analysis for colon cancer metabolomics: A case report. SPECIAL CARE IN DENTISTRY 2021; 42:80-85. [PMID: 34293207 DOI: 10.1111/scd.12635] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/18/2021] [Revised: 07/11/2021] [Accepted: 07/13/2021] [Indexed: 11/28/2022]
Abstract
BACKGROUND A key priority in colon cancer research is the identification of molecular biomarkers to improve early diagnosis, guide prognosis, and the design of new therapeutic approaches. Saliva is a powerful diagnostic biofluid that can be used to detect systemic alterations. This study aimed to investigate the parotid saliva (PS) metabolic Proton Nuclear Magnetic Resonance (1 H-NMR) profile of a patient diagnosed with colon cancer, and the subsequent changes 1 year after the end of chemotherapy. CASE REPORT We describe the 1 H-NMR PS spectrum of a 65-year-old woman diagnosed with colon cancer (G3 pT3 pN1c) (T0), and the changes in the spectrum from PS collected 1 year after the end of chemotherapy (XELOX: capecitabine plus oxaliplatin) (T1). The data was co-analyzed with blood test cancer antigens (S-CEA; S-CA19-9) and thyroid peroxidase antibody (TPOAb) measurements obtained simultaneously in order to identify peaks and interpret the spectra. The blood cancer antigens (S-CEA; S-CA19-9) and the PS 1 H-NMR peaks for fatty acids, lactate, acetate, N-acetyl sugars, citrate, tyrosine, saccharides, and formate decreased at T1 compared to T0. Whereas, the thyroid peroxidase antibody (TPOAb) blood values increased at T1 compared to T0 reflecting the changes in the 1 H-NMR spectral window of 1-3.5 ppm. CONCLUSION PS 1 H-NMR profiling identified modified metabolites that revealed cancer cells metabolism disturbances that subsequently decreased with time throughout treatment. These altered metabolites are potential biomarkers, providing a molecular diagnostic approach for clinical diagnosis, and prognosis of human colon cancer.
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Affiliation(s)
- Angela Rovera
- Dental Physical Sciences Unit, Centre for Oral Bioengineering, Institute of Dentistry, Queen Mary University of London, London, UK
| | - Mark Hector
- School of Dentistry, University of Dundee, Dundee, UK
| | - Paul Anderson
- Dental Physical Sciences Unit, Centre for Oral Bioengineering, Institute of Dentistry, Queen Mary University of London, London, UK
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34
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Ni D, Smyth HE, Gidley MJ, Cozzolino D. Towards personalised saliva spectral fingerprints: Comparison of mid infrared spectra of dried and whole saliva samples. SPECTROCHIMICA ACTA. PART A, MOLECULAR AND BIOMOLECULAR SPECTROSCOPY 2021; 253:119569. [PMID: 33610099 DOI: 10.1016/j.saa.2021.119569] [Citation(s) in RCA: 4] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 01/06/2021] [Revised: 01/27/2021] [Accepted: 01/28/2021] [Indexed: 06/12/2023]
Abstract
The aims of this study were to compare two sample presentations (dry and whole) as well as the effects of both gender and age on the mid infrared (MIR) fingerprint spectra of human saliva. Unstimulated saliva was collected from 52 Female (31 subjects, aged 40.9 ± 14.6 year) and Male (21 subjects, aged 34 ± 11.8 year) participants, stored frozen, and subsequently thawed and analysed by MIR spectroscopy as whole and dried saliva, respectively. Data were analysed by means of principal components analysis (PCA) and partial least squares (PLS) to interpret and compare the effects of presentation (dry vs whole), age and gender on the MIR spectra of saliva. Interpretation of the MIR spectra of both whole and dried samples revealed specific characteristic and different spectral signals when gender and age were compared in the amide I and amide II of proteins (e.g. albumin) and DNA. While whole saliva analysis might be more convenient for rapid test, dried saliva spectra were more consistent across replicates, demonstrating greater ability to distinguish individual differences. The interpretation of the PCA and PLS loadings of both whole and dried saliva samples allowed identification of specific MIR regions associated with age and gender of participants between 1000 cm-1 and 1800 cm-1. In particular, the MIR regions associated with the absorption of polysaccharides, glycosylated proteins, and nucleic acid phosphate groups present in saliva were the most dominant. This paper demonstrates that MIR spectroscopy can be used to measure saliva samples and to interpret individual differences in participants due to age in either dry or whole samples. No clear trends were observed in the MIR spectra of the samples associated with gender when all samples were analysed together. However, PLS regression models were able to predict gender in a subset of samples having similar age. The approach described in this study shows promise for potentially using saliva as a tool in food studies (e.g. saliva interactions between food and consumers).
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Affiliation(s)
- Dongdong Ni
- Centre for Nutrition and Food Sciences, Queensland Alliance for Agriculture and Food Innovation, The University of Queensland, St Lucia 4072, Queensland, Australia
| | - Heather E Smyth
- Centre for Nutrition and Food Sciences, Queensland Alliance for Agriculture and Food Innovation, The University of Queensland, St Lucia 4072, Queensland, Australia
| | - Michael J Gidley
- Centre for Nutrition and Food Sciences, Queensland Alliance for Agriculture and Food Innovation, The University of Queensland, St Lucia 4072, Queensland, Australia
| | - Daniel Cozzolino
- Centre for Nutrition and Food Sciences, Queensland Alliance for Agriculture and Food Innovation, The University of Queensland, St Lucia 4072, Queensland, Australia.
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35
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Wu D, Carillo KJ, Shie JJ, Yu SSF, Tzou DLM. Resolving Entangled JH-H-Coupling Patterns for Steroidal Structure Determinations by NMR Spectroscopy. Molecules 2021; 26:molecules26092643. [PMID: 33946512 PMCID: PMC8124291 DOI: 10.3390/molecules26092643] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/06/2021] [Revised: 04/28/2021] [Accepted: 04/29/2021] [Indexed: 11/16/2022] Open
Abstract
For decades, high-resolution 1H NMR spectroscopy has been routinely utilized to analyze both naturally occurring steroid hormones and synthetic steroids, which play important roles in regulating physiological functions in humans. Because the 1H signals are inevitably superimposed and entangled with various JH–H splitting patterns, such that the individual 1H chemical shift and associated JH–H coupling identities are hardly resolved. Given this, applications of thess information for elucidating steroidal molecular structures and steroid/ligand interactions at the atomic level were largely restricted. To overcome, we devoted to unraveling the entangled JH–H splitting patterns of two similar steroidal compounds having fully unsaturated protons, i.e., androstanolone and epiandrosterone (denoted as 1 and 2, respectively), in which only hydroxyl and ketone substituents attached to C3 and C17 were interchanged. Here we demonstrated that the JH–H values deduced from 1 and 2 are universal and applicable to other steroids, such as testosterone, 3β, 21-dihydroxygregna-5-en-20-one, prednisolone, and estradiol. On the other hand, the 1H chemical shifts may deviate substantially from sample to sample. In this communication, we propose a simple but novel scheme for resolving the complicate JH–H splitting patterns and 1H chemical shifts, aiming for steroidal structure determinations.
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Affiliation(s)
- Danni Wu
- Institute of Chemistry, Academia Sinica, Nankang, Taipei 11529, Taiwan; (D.W.); (K.J.C.); (J.-J.S.); (S.S.-F.Y.)
| | - Kathleen Joyce Carillo
- Institute of Chemistry, Academia Sinica, Nankang, Taipei 11529, Taiwan; (D.W.); (K.J.C.); (J.-J.S.); (S.S.-F.Y.)
- International Graduate Program, SCST, Academia Sinica, Nankang, Taipei 11529, Taiwan
- The Department of Applied Chemistry, National Yang Ming Chiao Tung University, Hsinchu 30013, Taiwan
| | - Jiun-Jie Shie
- Institute of Chemistry, Academia Sinica, Nankang, Taipei 11529, Taiwan; (D.W.); (K.J.C.); (J.-J.S.); (S.S.-F.Y.)
| | - Steve S.-F. Yu
- Institute of Chemistry, Academia Sinica, Nankang, Taipei 11529, Taiwan; (D.W.); (K.J.C.); (J.-J.S.); (S.S.-F.Y.)
| | - Der-Lii M. Tzou
- Institute of Chemistry, Academia Sinica, Nankang, Taipei 11529, Taiwan; (D.W.); (K.J.C.); (J.-J.S.); (S.S.-F.Y.)
- Department of Applied Chemistry, National Chia-Yi University, Chia-Yi 60004, Taiwan
- Correspondence:
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Kocatürk M, Tecles F, Yalçın E, Cihan H, Tural M, Levent P, Cansev M, Cerón JJ, Yilmaz Z. Changes in choline and cholinesterase in saliva of dogs with parvovirus infection. Res Vet Sci 2020; 134:147-149. [PMID: 33385977 DOI: 10.1016/j.rvsc.2020.12.012] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/11/2020] [Revised: 12/09/2020] [Accepted: 12/22/2020] [Indexed: 11/30/2022]
Abstract
This study investigated the changes in choline (Ch) and butyrylcholinesterase (BChE) in saliva in canine parvovirosis (CP) as a model of sepsis, and their correlations with these analytes in serum and with other markers of inflammation such as white blood cell count (WBC) and serum C-reactive protein (CRP). A total of 30 dogs with CP were sampled for saliva and serum at presentation, and 10 healthy puppies were also sampled as controls. Salivary Ch was higher in dogs with CP (P < 0.001) showing a positive correlation with CRP, whereas no differences were observed in salivary BChE. This is the first report in which Ch is measured in saliva of dogs and based in the results of this study, salivary Ch could be potentially used as biomarker of the severity of CP.
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Affiliation(s)
- Meriç Kocatürk
- Department of Internal Medicine, Faculty of Veterinary Medicine, Bursa Uludag University, 16059 Bursa, Turkey
| | - Fernando Tecles
- Interdisciplinary Laboratory of Clinical Analysis, Interlab-UMU, Regional Campus of International Excellence 'Campus Mare Nostrum', University of Murcia, 30100 Murcia, Spain.
| | - Ebru Yalçın
- Department of Internal Medicine, Faculty of Veterinary Medicine, Bursa Uludag University, 16059 Bursa, Turkey
| | - Hüseyin Cihan
- Department of Internal Medicine, Faculty of Veterinary Medicine, Bursa Uludag University, 16059 Bursa, Turkey
| | - Merve Tural
- Department of Internal Medicine, Faculty of Veterinary Medicine, Bursa Uludag University, 16059 Bursa, Turkey
| | - Pinar Levent
- Department of Internal Medicine, Faculty of Veterinary Medicine, Bursa Uludag University, 16059 Bursa, Turkey
| | - Mehmet Cansev
- Department of Pharmacology, Faculty of Medicine, Bursa Uludag University, 16059 Bursa, Turkey
| | - José J Cerón
- Interdisciplinary Laboratory of Clinical Analysis, Interlab-UMU, Regional Campus of International Excellence 'Campus Mare Nostrum', University of Murcia, 30100 Murcia, Spain
| | - Zeki Yilmaz
- Department of Internal Medicine, Faculty of Veterinary Medicine, Bursa Uludag University, 16059 Bursa, Turkey
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Duarte D, Castro B, Pereira JL, Marques JF, Costa AL, Gil AM. Evaluation of Saliva Stability for NMR Metabolomics: Collection and Handling Protocols. Metabolites 2020; 10:E515. [PMID: 33352779 PMCID: PMC7766053 DOI: 10.3390/metabo10120515] [Citation(s) in RCA: 17] [Impact Index Per Article: 3.4] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/30/2020] [Revised: 12/14/2020] [Accepted: 12/16/2020] [Indexed: 12/15/2022] Open
Abstract
Maintaining a salivary metabolic profile upon sample collection and preparation is determinant in metabolomics. Nuclear magnetic resonance (NMR) spectroscopy was used to identify metabolite changes during short-term storage, at room temperature (RT)/4 °C/-20 °C, and after sample preparation, at RT/4 °C (mimicking typical clinical/laboratory settings). Interestingly, significant metabolic inter-individual and inter-day variability were noted, probably determining sample stability to some extent. After collection, no changes were noted at -20 °C (at least for 4 weeks). RT storage induced decreases in methylated macromolecules (6 h); lactate (8 h); alanine (12 h); galactose, hypoxanthine, pyruvate (24 h); sarcosine, betaine, choline, N-acetyl-glycoproteins (48 h), while acetate increased (48 h). Less, but different, changes were observed at 4 °C, suggesting different oral and microbial status at different temperatures (with a possible contribution from inter-individual and inter-day variability), and identifying galactose, hypoxanthine, and possibly, choline esters, as potential general stability indicators. After preparation, addition of NaN3 did not impact significantly on saliva stabilization, neither at RT nor at 4 °C, although its absence was accompanied by slight increases in fucose (6.5 h) and proline (8 h) at RT, and in xylose (24 h) at 4 °C. The putative metabolic origins of the above variations are discussed, with basis on the salivary microbiome. In summary, after collection, saliva can be stored at RT/4 °C for up to 6 h and at -20 °C for at least 4 weeks. Upon preparation for NMR analysis, samples are highly stable at 25 °C up to 8 h and at 4 °C up to 48 h, with NaN3 addition preventing possible early changes in fucose, proline (6-8 h), and xylose (24 h) levels.
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Affiliation(s)
- Daniela Duarte
- CICECO—Department of Chemistry, Aveiro Institute of Materials, Campus Universitário de Santiago, University of Aveiro, 3810-193 Aveiro, Portugal; (D.D.); (B.C.)
| | - Beatriz Castro
- CICECO—Department of Chemistry, Aveiro Institute of Materials, Campus Universitário de Santiago, University of Aveiro, 3810-193 Aveiro, Portugal; (D.D.); (B.C.)
| | - Joana Leonor Pereira
- Dentistry Department, Faculty of Medicine, Institute of Paediatric and Preventive Dentistry, University of Coimbra, 3000-075 Coimbra, Portugal; (J.L.P.); (A.L.C.)
| | - Joana Faria Marques
- GIBBO-Oral Biology and Biochemistry Research Group, CEMBDE-COCHRANE Portugal—Faculty of Dental Medicine, Universidade de Lisboa, 1649-003 Lisboa, Portugal;
| | - Ana Luísa Costa
- Dentistry Department, Faculty of Medicine, Institute of Paediatric and Preventive Dentistry, University of Coimbra, 3000-075 Coimbra, Portugal; (J.L.P.); (A.L.C.)
| | - Ana M. Gil
- CICECO—Department of Chemistry, Aveiro Institute of Materials, Campus Universitário de Santiago, University of Aveiro, 3810-193 Aveiro, Portugal; (D.D.); (B.C.)
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Costa dos Santos Junior G, Pereira CM, Kelly da Silva Fidalgo T, Valente AP. Saliva NMR-Based Metabolomics in the War Against COVID-19. Anal Chem 2020; 92:15688-15692. [PMID: 33215503 PMCID: PMC7688045 DOI: 10.1021/acs.analchem.0c04679] [Citation(s) in RCA: 13] [Impact Index Per Article: 2.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/06/2020] [Accepted: 11/10/2020] [Indexed: 12/15/2022]
Abstract
COVID-19 is an emergent, worldwide public health concern. Joint efforts have been made by scientific communities of various fields to better understand the mechanisms of action of SARS-CoV-2. The need to understand the pathophysiological fingerprint and pathways of this disease make metabolomics-related approaches an indispensable tool for properly answering concerns relating to disease course. Determination of the metabolomic profile may help to explain the heterogeneous spectra of COVID-19 clinical phenotypes and be useful in monitoring disease progression as well as therapeutic treatments. In this sense, saliva has proven to be a strategic biofluid, owing not only to its appeal as a noninvasive sampling method but also due to the capacity of the virus to invade epithelial cells of the oral mucosa and salivary gland ducts via ACE2 receptors. Accordingly, important changes in metabolism have been described relating to COVID-19, indicating that metabolomics may open new avenues for understanding the pathophysiology of this disease, especially via longitudinal study designs. Thus, we discuss the importance of comprehending the SARS-CoV-2 salivary metabolomic fingerprint and also highlight the situation of Brazil on the frontlines of the war against COVID-19.
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Affiliation(s)
- Gilson Costa dos Santos Junior
- Laboratory of NMR Metabolomics, IBRAG, Department of
Genetics, State University of Rio de Janeiro, Boulevard 28 de
Setembro 77 fds, Vila Isabel, RJ, 20551-030 Rio de Janeiro,
Brazil
| | - Claudia Maria Pereira
- Postgraduate Program in Translational Biomedicine,
Grande Rio University, Rua Professor José de Souza
Herdy, 1160, Jardim Vinte e Cinco de Agosto, 25071-202 Duque de Caxias,
Brazil
| | - Tatiana Kelly da Silva Fidalgo
- Dental School, Department of Preventive and Community
Dentistry, State University of Rio de Janeiro, Boulevard 28 de
Setembro, 157, Vila Isabel, 20551-030 Rio de Janeiro, Brazil
| | - Ana Paula Valente
- BioNMR, CENABIO I, Department of Structural Biology,
Federal University of Rio de Janeiro, Av. Carlos Chagas
Filho, 373, CCS/bloco K-anexo, 21941-599 Rio de Janeiro, Brazil
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Changes in the Salivary Metabolic Profile of Generalized Periodontitis Patients after Non-surgical Periodontal Therapy: A Metabolomic Analysis Using Nuclear Magnetic Resonance Spectroscopy. J Clin Med 2020; 9:jcm9123977. [PMID: 33302593 PMCID: PMC7763572 DOI: 10.3390/jcm9123977] [Citation(s) in RCA: 10] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/29/2020] [Revised: 11/29/2020] [Accepted: 12/03/2020] [Indexed: 12/11/2022] Open
Abstract
Pattern analysis of the salivary metabolic profile has been proven accurate in discriminating between generalized periodontitis (GP) patients and healthy individuals (HI), as this disease modifies the salivary concentrations of specific metabolites. Due to the scarcity of data from previous studies, this study aimed to evaluate if non-surgical periodontal therapy (NST) could affect the metabolomic profile in GP patients’ saliva and if it compares to that of HI. Unstimulated salivary samples were collected from 11 HI and 12 GP patients before and 3 months after NST. Nuclear Magnetic Resonance (NMR) spectroscopy, followed by a supervised multivariate statistical approach on entire saliva spectra and partial least square (PLS) discriminant analysis, were performed to obtain metabolic profiles. In the GP group, periodontal treatment improved all clinical parameters, but not all the diseased sites were eradicated. PLS revealed an accuracy of 100% in distinguishing between metabolic profiles of GP patients before and after NST. Orthogonal projection to latent structure was able to discriminate between the three groups of subjects with an accuracy of 85.6%. However, the post-NST metabolic profile of GP patients could not be completely assimilated to that of HI. Although NST may produce significant changes in the metabolic profile, GP patients maintained a distinctive fingerprint compared to HI.
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Abstract
Introduction: Saliva is an ideal biofluid that can be collected in a noninvasive manner, enabling safe and frequent screening of various diseases. Recent studies have revealed that salivary metabolomics analysis has the potential to detect both oral and systemic cancers. Area covered: We reviewed the technical aspects, as well as applications, of salivary metabolomics for cancer detection. The topics include the effects of preconditioning and the method of sample collection, sample storage, processing, measurement, data analysis, and validation of the results. We also examined the rational relationship between salivary biomarkers and tumors distant from the oral cavity. A strategy to establish standard operating protocols for obtaining reproducible quantification data is also discussed Expert opinion: Salivary metabolomics reflects oral and systematic health status, which potently enables cancer detection. The sensitivity and specificity of each marker and their combinations have been well evaluated, but a validation study is required. Further, the standard operating protocol for each procedure should be established to obtain reproducible data before clinical usage.
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Affiliation(s)
- Masahiro Sugimoto
- Research and Development Centre for Minimally Invasive Therapies, Medical Research Institute, Tokyo Medical University , Tokyo, Japan.,Institute for Advanced Biosciences, Keio University , Yamagata, Japan
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Herrala M, Mikkonen JJW, Pesonen P, Lappalainen R, Tjäderhane L, Niemelä RK, Seitsalo H, Salo T, Myllymaa S, Kullaa AM. Variability of salivary metabolite levels in patients with Sjögren's syndrome. J Oral Sci 2020; 63:22-26. [PMID: 33116006 DOI: 10.2334/josnusd.19-0504] [Citation(s) in RCA: 20] [Impact Index Per Article: 4.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/01/2022]
Abstract
PURPOSE To investigate inter- and intra-individual variation in the levels and outputs (concentration multiplied by salivary flow rate) of salivary metabolites in patients with primary Sjögren's syndrome (pSS). METHODS A total of 56 samples of stimulated saliva were collected from 14 female pSS patients during four laboratory visits within 20 weeks and analyzed using proton nuclear magnetic resonance spectroscopy. Single saliva samples from each of 15 controls were also analyzed. RESULTS Among 21 quantified metabolites, choline was significantly elevated in the pSS patients at each time point (P ≤ 0.015), taurine at the last three time points (P ≤ 0.013), alanine at the last two time points (P ≤ 0.007) and glycine at the last time point (P = 0.005). Inter-individual variation in metabolite concentrations was generally larger among the patients than among the controls, and significantly large variations were observed for glycine (P ≤ 0.007, all time points), choline (P ≤ 0.033, three last time points) and alanine (P = 0.028, baseline). Metabolite output analysis showed that choline had the lowest intra-patient variation. CONCLUSION In spite of considerable intra- and inter-individual variation, levels and outputs of specific metabolites in patients with pSS differ from those in controls, and may be potentially applicable as new biological markers for monitoring of the response to treatment.
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Affiliation(s)
- Maria Herrala
- Research Group of Oral Health Sciences, Faculty of Medicine, University of Oulu and Oulu University Hospital.,Institute of Dentistry, Faculty of Health Sciences, University of Eastern Finland.,Medical Research Center Oulu, University of Oulu and Oulu University Hospital
| | - Jopi J W Mikkonen
- SIB Labs and Department of Applied Physics, Faculty of Science and Forestry, University of Eastern Finland
| | - Paula Pesonen
- Infrastructure for Population Studies, Faculty of Medicine, University of Oulu
| | - Reijo Lappalainen
- SIB Labs and Department of Applied Physics, Faculty of Science and Forestry, University of Eastern Finland
| | - Leo Tjäderhane
- Medical Research Center Oulu, University of Oulu and Oulu University Hospital.,Department of Oral and Maxillofacial Diseases, University of Helsinki and Helsinki University Hospital
| | | | | | - Tuula Salo
- Medical Research Center Oulu, University of Oulu and Oulu University Hospital.,Department of Oral and Maxillofacial Diseases, University of Helsinki and Helsinki University Hospital
| | - Sami Myllymaa
- SIB Labs and Department of Applied Physics, Faculty of Science and Forestry, University of Eastern Finland
| | - Arja M Kullaa
- Research Group of Oral Health Sciences, Faculty of Medicine, University of Oulu and Oulu University Hospital.,Institute of Dentistry, Faculty of Health Sciences, University of Eastern Finland.,Educational Dental Clinic, Kuopio University Hospital
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1H-NMR-Based salivary metabolomics from females with temporomandibular disorders - A pilot study. Clin Chim Acta 2020; 510:625-632. [PMID: 32791140 DOI: 10.1016/j.cca.2020.08.006] [Citation(s) in RCA: 8] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/12/2020] [Revised: 08/01/2020] [Accepted: 08/04/2020] [Indexed: 12/16/2022]
Abstract
Although temporomandibular disorder (TMD) is the second most common musculoskeletal disorder in the general population, the disease is multifactorial and presents symptoms common to other conditions which misdiagnosis can lead to treatment failure. In this case-control study, we performed, for the first time, a high-resolution 1H-nuclear magnetic resonance spectroscopy metabolomic analysis of the saliva of 26 women with TMD of muscular origin (experimental group [EG]) at the beginning (EG-pre) and at the end (EG-post) of a conservative treatment, and of 27 normal women (control group [CG]) to identify a metabolic signature for TMD. One-way analysis of variance showed changes in the concentration of phenylacetate, dimethylamine, maltose, acetoin, and isovalerate. Partial least-square discriminant analysis showed that metabolite signals did not overlap in CG X EG-pre and EG-pre X EG-post, but overlapped in CG X EG-post. The area under the receiver operating characteristic curve was 1 in CG X EG-pre (95% CI, 1.000-1.000; p < 0.002), 0.993 in EG-pre X EG-post (95% CI, 0.963-1.000), and 0.832 in CG X EG-post (95% CI, 0.699-0.961). These results suggest that the metabolomic profiles of women with and without TMD differ, while after treatment there is a lower distinction and slight tendency towards overlapping between CG and EG-post compared to pre treatment. We also found that phenylacetate, dimethylamine, maltose, acetoin, and isovalerate are potential biomarkers for TMD of muscular origin.
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Meleti M, Quartieri E, Antonelli R, Pezzi ME, Ghezzi B, Viani MV, Setti G, Casali E, Ferrari E, Ciociola T, Spisni A, Pertinhez TA. Metabolic Profiles of Whole, Parotid and Submandibular/Sublingual Saliva. Metabolites 2020; 10:E318. [PMID: 32781584 PMCID: PMC7466076 DOI: 10.3390/metabo10080318] [Citation(s) in RCA: 19] [Impact Index Per Article: 3.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/15/2020] [Revised: 07/31/2020] [Accepted: 08/05/2020] [Indexed: 01/05/2023] Open
Abstract
The detection of salivary molecules associated with pathological and physiological alterations has encouraged the search of novel and non-invasive diagnostic biomarkers for oral health evaluation. While genomic, transcriptomic, and proteomic profiles of human saliva have been reported, its metabolic composition is a topic of research: metabolites in submandibular/sublingual saliva have never been analyzed systematically. In this study, samples of whole, parotid, and submandibular/sublingual saliva from 20 healthy donors, without dental or periodontal diseases, were examined by nuclear magnetic resonance. We identified metabolites which are differently distributed within the three saliva subtypes (54 in whole, 49 in parotid, and 36 in submandibular/sublingual saliva). Principal component analysis revealed a distinct cluster for whole saliva and a partial overlap for parotid and submandibular/sublingual metabolites. We found exclusive metabolites for each subtype: 2-hydroxy-3-methylvalerate, 3-methyl-glutarate, 3-phenylpropionate, 4-hydroxyphenylacetate, 4-hydroxyphenyllactate, galactose, and isocaproate in whole saliva; caprylate and glycolate in submandibular/sublingual saliva; arginine in parotid saliva. Salivary metabolites were classified into standard and non-proteinogenic amino acids and amines; simple carbohydrates; organic acids; bacterial-derived metabolites. The identification of a salivary gland-specific metabolic composition in healthy people provides the basis to invigorate the search for salivary biomarkers associated with oral and systemic diseases.
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Affiliation(s)
- Marco Meleti
- Centro Universitario Odontoiatria, University of Parma, Via Gramsci 14, 43126 Parma, Italy; (M.M.); (R.A.); (M.E.P.); (B.G.); (M.V.V.); (G.S.)
- Department of Medicine and Surgery, University of Parma, Via Volturno 39, 43125 Parma, Italy; (E.Q.); (E.C.); (T.C.); (A.S.); (T.A.P.)
| | - Eleonora Quartieri
- Department of Medicine and Surgery, University of Parma, Via Volturno 39, 43125 Parma, Italy; (E.Q.); (E.C.); (T.C.); (A.S.); (T.A.P.)
- Transfusion Medicine Unit, Azienda USL-IRCCS di Reggio Emilia, Viale Umberto I 50, 43123 Reggio Emilia, Italy
| | - Rita Antonelli
- Centro Universitario Odontoiatria, University of Parma, Via Gramsci 14, 43126 Parma, Italy; (M.M.); (R.A.); (M.E.P.); (B.G.); (M.V.V.); (G.S.)
| | - Margherita E. Pezzi
- Centro Universitario Odontoiatria, University of Parma, Via Gramsci 14, 43126 Parma, Italy; (M.M.); (R.A.); (M.E.P.); (B.G.); (M.V.V.); (G.S.)
| | - Benedetta Ghezzi
- Centro Universitario Odontoiatria, University of Parma, Via Gramsci 14, 43126 Parma, Italy; (M.M.); (R.A.); (M.E.P.); (B.G.); (M.V.V.); (G.S.)
| | - Maria Vittoria Viani
- Centro Universitario Odontoiatria, University of Parma, Via Gramsci 14, 43126 Parma, Italy; (M.M.); (R.A.); (M.E.P.); (B.G.); (M.V.V.); (G.S.)
| | - Giacomo Setti
- Centro Universitario Odontoiatria, University of Parma, Via Gramsci 14, 43126 Parma, Italy; (M.M.); (R.A.); (M.E.P.); (B.G.); (M.V.V.); (G.S.)
| | - Emanuela Casali
- Department of Medicine and Surgery, University of Parma, Via Volturno 39, 43125 Parma, Italy; (E.Q.); (E.C.); (T.C.); (A.S.); (T.A.P.)
| | - Elena Ferrari
- Department of Medicine and Surgery, University of Parma, Via Volturno 39, 43125 Parma, Italy; (E.Q.); (E.C.); (T.C.); (A.S.); (T.A.P.)
| | - Tecla Ciociola
- Department of Medicine and Surgery, University of Parma, Via Volturno 39, 43125 Parma, Italy; (E.Q.); (E.C.); (T.C.); (A.S.); (T.A.P.)
| | - Alberto Spisni
- Department of Medicine and Surgery, University of Parma, Via Volturno 39, 43125 Parma, Italy; (E.Q.); (E.C.); (T.C.); (A.S.); (T.A.P.)
| | - Thelma A. Pertinhez
- Department of Medicine and Surgery, University of Parma, Via Volturno 39, 43125 Parma, Italy; (E.Q.); (E.C.); (T.C.); (A.S.); (T.A.P.)
- Transfusion Medicine Unit, Azienda USL-IRCCS di Reggio Emilia, Viale Umberto I 50, 43123 Reggio Emilia, Italy
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Khurshid Z, Warsi I, Moin SF, Slowey PD, Latif M, Zohaib S, Zafar MS. Biochemical analysis of oral fluids for disease detection. Adv Clin Chem 2020; 100:205-253. [PMID: 33453866 DOI: 10.1016/bs.acc.2020.04.005] [Citation(s) in RCA: 13] [Impact Index Per Article: 2.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/07/2023]
Abstract
The field of diagnostics using invasive blood testing represents the majority of diagnostic tests used as part of routine health monitoring. The relatively recent introduction of salivary diagnostics has lead to a major paradigm shift in diagnostic analyses. Additionally, in this era of big data, oral fluid testing has shown promising outcomes in a number of fields, particularly the areas of genomics, microbiomics, proteomics, metabolomics, and transcriptomics. Despite the analytical challenges involved in the interpretation of large datasets generated from biochemical studies involving bodily fluids, including saliva, many studies have identified novel oral biomarkers for diagnosing oral and systemic diseases. In this regard, oral biofluids, including saliva, gingival crevicular fluid (GCF), peri-implant crevicular fluid (PICF), dentinal tubular fluid (DTF), are now attracting increasing attention due to their important attributes, such as noninvasive sampling, easy handling, low cost, and more accurate diagnosis of oral diseases. Recently, the utilization of salivary diagnostics to evaluate systemic diseases and monitor general health has increased in popularity among clinicians. Saliva contains a wide range of protein, DNA and RNA biomarkers, which assist in the diagnosis of multiple diseases and conditions, including cancer, cardiovascular diseases (CVD), auto-immune and degenerative diseases, respiratory infections, oral diseases, and microbial (viral, bacterial and fungal) diseases. Moreover, due to its noninvasive nature and ease-of-adoption by children, it is now being used in mass screening programs, oral health-related studies and clinical trials in support of the development of therapeutic agents. The recent advent of highly sensitive technologies, such as next-generation sequencing, mass spectrometry, highly sensitives ELISAs, and homogeneous immunoassays, suggests that even small quantities of salivary biomarkers are able to be assayed accurately, providing opportunities for the development of many future diagnostic applications (including emerging technologies, such as point-of-care and rapid molecular technologies). The present article explores the omics and biochemical compositions of various oral biofluids with important value in diagnostics and monitoring.
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Affiliation(s)
- Zohaib Khurshid
- Department of Prosthodontics and Dental Implantology, College of Dentistry, King Faisal University, Al-Ahsa, Saudi Arabia
| | - Ibrahim Warsi
- Masters in Medical Science and Clinical Investigation, Harvard Medical School, Boston, MA, United States
| | - Syed F Moin
- National Center for Proteomics, University of Karachi, Karachi, Pakistan
| | - Paul D Slowey
- Oasis Diagnostics® Corporation, Vancouver, WA, United States
| | - Muhammad Latif
- Centre for Genetics and Inherited Diseases (CGID), Taibah University, Al Madinah Al Munawwarah, Saudi Arabia
| | - Sana Zohaib
- Department of Biomedical Engineering, College of Engineering, King Faisal University, Al-Ahsa, Saudi Arabia
| | - Muhammad S Zafar
- Department of Restorative Dentistry, College of Dentistry, Taibah University, Al Madinah Al Munawwarah, Saudi Arabia; Department of Dental Materials, Islamic International Dental College, Riphah International University, Islamabad, Pakistan.
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45
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Abstract
The oral microbiome is one of the most stable ecosystems in the body and yet the reasons for this are still unclear. As well as being stable, it is also highly diverse which can be ascribed to the variety of niches available in the mouth. Previous studies have focused on the microflora in disease-either caries or periodontitis-and only recently have they considered factors that maintain the normal microflora. This has led to the perception that the microflora proliferate in nutrient-rich periods during oral processing of foods and drinks and starves in between times. In this review, evidence is presented which shows that the normal flora are maintained on a diet of salivary factors including urea, lactate, and salivary protein degradation. These factors are actively secreted by salivary glands which suggests these factors are important in maintaining normal commensals in the mouth. In addition, the immobilization of SIgA in the mucosal pellicle indicates a mechanism to retain certain bacteria that does not rely on the bacterial-centric mechanisms such as adhesins. By examining the salivary metabolome, it is clear that protein degradation is a key nutrient and the availability of free amino acids increases resistance to environmental stresses.
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Affiliation(s)
- G H Carpenter
- Salivary Research, Centre for Host-microbiome Interactions, Faculty of Dental, Oral & Craniofacial Sciences, King's College London, London, UK
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46
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Abstract
Metabolomics has been identified as a means of functionally assessing the net biological activity of a particular microbial community. Considering the oral microbiome, such an approach remains largely underused. While the current knowledge of the oral microbiome is constantly expanding, there are several deficits in knowledge particularly relating to their interactions with their host. This work uses nuclear magnetic resonance spectroscopy to investigate metabolic differences between oral microbial metabolism of endogenous (i.e., salivary protein) and exogenous (i.e., dietary carbohydrates) substrates. It also investigated whether microbial generation of different metabolites may be associated with host taste perception. This work found that in the absence of exogenous substrate, oral bacteria readily catabolize salivary protein and generate metabolic profiles similar to those seen in vivo. Important metabolites such as acetate, butyrate, and propionate are generated at relatively high concentrations. Higher concentrations of metabolites were generated by tongue biofilm compared to planktonic salivary bacteria. Thus, as has been postulated, metabolite production in proximity to taste receptors could reach relatively high concentrations. In the presence of 0.25 M exogenous sucrose, increased catabolism was observed with increased concentrations of a range of metabolites relating to glycolysis (lactate, pyruvate, succinate). Additional pyruvate-derived molecules such as acetoin and alanine were also increased. Furthermore, there was evidence that individual taste sensitivity to sucrose was related to differences in the metabolic fate of sucrose in the mouth. High-sensitivity perceivers appeared more inclined toward continual citric acid cycle activity postsucrose, whereas low-sensitivity perceivers had a more efficient conversion of pyruvate to lactate. This work collectively indicates that the oral microbiome exists in a complex balance with the host, with fluctuating metabolic activity depending on nutrient availability. There is preliminary evidence of an association between host behavior (sweet taste perception) and oral catabolism of sugar.
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Affiliation(s)
- A Gardner
- Salivary Research, Centre for Host-Microbiome Interactions, Faculty of Dental, Oral & Craniofacial Sciences, King's College London, London, UK
- Department of Restorative Dentistry, Dental Hospital and School, University of Dundee, Dundee, UK
| | - P W So
- Department of Neuroimaging, Institute of Psychiatry, Psychology and Neuroscience, King's College London, Maurice Wohl Clinical Neuroscience Institute, London, UK
| | - G H Carpenter
- Salivary Research, Centre for Host-Microbiome Interactions, Faculty of Dental, Oral & Craniofacial Sciences, King's College London, London, UK
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Correlation between parotid saliva composition and dental caries using 31P-NMR and ICDAS score. Arch Oral Biol 2020; 111:104651. [DOI: 10.1016/j.archoralbio.2020.104651] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/04/2019] [Revised: 01/02/2020] [Accepted: 01/04/2020] [Indexed: 11/18/2022]
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Gardner A, Carpenter G, So PW. Salivary Metabolomics: From Diagnostic Biomarker Discovery to Investigating Biological Function. Metabolites 2020; 10:E47. [PMID: 31991929 PMCID: PMC7073850 DOI: 10.3390/metabo10020047] [Citation(s) in RCA: 89] [Impact Index Per Article: 17.8] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/13/2019] [Revised: 01/15/2020] [Accepted: 01/24/2020] [Indexed: 12/12/2022] Open
Abstract
Metabolomic profiling of biofluids, e.g., urine, plasma, has generated vast and ever-increasing amounts of knowledge over the last few decades. Paradoxically, metabolomic analysis of saliva, the most readily-available human biofluid, has lagged. This review explores the history of saliva-based metabolomics and summarizes current knowledge of salivary metabolomics. Current applications of salivary metabolomics have largely focused on diagnostic biomarker discovery and the diagnostic value of the current literature base is explored. There is also a small, albeit promising, literature base concerning the use of salivary metabolomics in monitoring athletic performance. Functional roles of salivary metabolites remain largely unexplored. Areas of emerging knowledge include the role of oral host-microbiome interactions in shaping the salivary metabolite profile and the potential roles of salivary metabolites in oral physiology, e.g., in taste perception. Discussion of future research directions describes the need to begin acquiring a greater knowledge of the function of salivary metabolites, a current research direction in the field of the gut metabolome. The role of saliva as an easily obtainable, information-rich fluid that could complement other gastrointestinal fluids in the exploration of the gut metabolome is emphasized.
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Affiliation(s)
- Alexander Gardner
- Salivary Research, Centre for Host–Microbiome Interactions, Faculty of Dental, Oral & Craniofacial Sciences, King’s College London, London SE1 9RT, UK; (A.G.); (G.C.)
- Department of Restorative Dentistry, Dental Hospital and School, University of Dundee, Dundee DD1 4HR, UK
| | - Guy Carpenter
- Salivary Research, Centre for Host–Microbiome Interactions, Faculty of Dental, Oral & Craniofacial Sciences, King’s College London, London SE1 9RT, UK; (A.G.); (G.C.)
| | - Po-Wah So
- Department of Neuroimaging, Institute of Psychiatry, Psychology and Neuroscience, King’s College London, Maurice Wohl Clinical Neuroscience Institute, London SE5 9RT, UK
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Gardner A, So P, Carpenter G. Endogenous salivary citrate is associated with enhanced rheological properties following oral capsaicin stimulation. Exp Physiol 2020; 105:96-107. [PMID: 31705555 PMCID: PMC6973168 DOI: 10.1113/ep088166] [Citation(s) in RCA: 12] [Impact Index Per Article: 2.4] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/10/2019] [Accepted: 11/04/2019] [Indexed: 12/14/2022]
Abstract
NEW FINDINGS What is the central question of this study? What are the relationships between physical properties of saliva, protein composition and metabolite composition? What is the main finding and its importance? Salivary citrate, one of the major endogenous metabolites in saliva, increased upon capsaicin stimulation and was associated with improved physical properties measured by extensional rheology. This suggests salivary gland citrate transporters might be a valuable area of future study. ABSTRACT Saliva displays viscoelastic properties which enable coating, lubrication and protection of the oral mucosa and hard tissues. Individuals lacking saliva or perceiving oral dryness can manage their symptoms using artificial saliva preparations, but these often fail to mimic the sensation and functionality of natural saliva. It is widely acknowledged that mucins (MUC7 and MUC5B) confer saliva's rheological properties, but artificial saliva containing purified mucins is still often an inadequate substitute. This work aimed to explore salivary components that influence salivary extensional rheology to better understand how natural saliva could be replicated. Saliva was stimulated via control and capsaicin solutions in healthy volunteers. Extensional rheology was analysed using a CaBER-1 (capillary breakup) extensional rheometer. Protein composition, including mucins, was measured by gel-electrophoresis band densitometry and metabolites were measured by 1 H nuclear magnetic resonance spectroscopy. Capsaicin stimulation significantly increased capillary breakup time, extensional viscosity and the abundance of most major salivary proteins. Stimulation also increased salivary citrate and choline concentrations. Significant correlations were found between capillary breakup time and amylase (r = 0.67, P < 0.05), statherin (ρ = 0.66, P < 0.05) and citrate (ρ = 0.81, P < 0.01). The relationship between citrate and salivary rheology was subsequently investigated in vitro. These results suggest that citrate and non-mucin proteins are stronger predictors of salivary rheology than the more often studied mucin glycoproteins. Potential mechanisms are discussed and future work in this area could help formulate more effective saliva substitutes, more closely resembling natural saliva.
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Affiliation(s)
- Alexander Gardner
- Salivary Research, Centre for Host‐Microbiome InteractionsFaculty of Dental, Oral & Craniofacial SciencesKing's College LondonLondonUK
- Department of Restorative Dentistry, Dental Hospital and SchoolUniversity of DundeeDundeeUK
| | - Po‐Wah So
- Department of Neuroimaging, Institute of PsychiatryPsychology and Neuroscience, King's College LondonMaurice Wohl Clinical Neuroscience InstituteLondonUK
| | - Guy Carpenter
- Salivary Research, Centre for Host‐Microbiome InteractionsFaculty of Dental, Oral & Craniofacial SciencesKing's College LondonLondonUK
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Singh MP, Saxena M, Saimbi CS, Siddiqui MH, Roy R. Post-periodontal surgery propounds early repair salivary biomarkers by 1H NMR based metabolomics. Metabolomics 2019; 15:141. [PMID: 31612356 DOI: 10.1007/s11306-019-1593-3] [Citation(s) in RCA: 8] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 01/24/2019] [Accepted: 09/25/2019] [Indexed: 12/13/2022]
Abstract
INTRODUCTION Oral microflora is a well-orchestrated and acts as a sequential defense mechanism for any infection related to oral disease. Chronic periodontitis is a disease of a microbial challenge to symbiosis and homeostasis. Periodontal surgery is the most promising cure with repair process during periodontal regeneration. It has an encouraging outcome in terms of early recovery biomarkers. OBJECTIVE Saliva of periodontal surgery subjects with the chronic periodontitis have been evaluated by 1H NMR spectroscopy in search of possible early metabolic differences that could be obtained in order to see the eradication of disease which favours the symbiotic condition. METHOD The study employed 1H NMR spectroscopy on 176 human saliva samples in search of distinctive differences and their spectral data were further subjected to multivariate and quantitative analysis. RESULT The 1H NMR study of periodontal surgery samples shows clear demarcation and profound metabolic differences when compared with the diseased condition. Several metabolites such as lactate, ethanol, succinate, and glutamate were found to be of higher significance in periodontal surgery in contrast to chronic periodontitis subjects. The PLS-DA model of the studied group resulted in R2 of 0.83 and Q2 of 0.70. CONCLUSION Significant metabolites could be considered as early repair markers for chronic periodontitis disease as they are being restored to achieve symbiosis. The study, therefore, concluded the early recovery process of the diseased subjects with the restoration of possible metabolomic profile similar to the healthy controls.
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Affiliation(s)
- Manvendra Pratap Singh
- Centre of Biomedical Research, formerly Centre of Biomedical Magnetic Resonance (CBMR), Sanjay Gandhi Postgraduate Institute of Medical Sciences Campus, Rae Bareli Road, Lucknow, 226014, India
| | - Mona Saxena
- Department of Periodontics, Career Post Graduate Institute of Dental Sciences and Hospital, Lucknow, 226020, India
- Department of Biochemistry, Saraswati Medical College, Unnao, India
| | - Charanjit S Saimbi
- Department of Periodontics, Career Post Graduate Institute of Dental Sciences and Hospital, Lucknow, 226020, India.
| | | | - Raja Roy
- Centre of Biomedical Research, formerly Centre of Biomedical Magnetic Resonance (CBMR), Sanjay Gandhi Postgraduate Institute of Medical Sciences Campus, Rae Bareli Road, Lucknow, 226014, India.
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