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      Targeted metabolomics of pellicle and saliva in children with different caries activity

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          Abstract

          Pellicle is the initial proteinaceous layer that is formed almost instantaneously on all solid surfaces in the oral cavity. It is of essential relevance for any interactions and metabolism on the tooth surface. Up to now, there is no information on the metabolome of this structure. Accordingly, the present study aims to characterise the metabolomic profile of in-situ pellicle in children with different caries activity for the first time in comparison to saliva. Small molecules such as carbohydrates, amino acids, organic acids, and fatty acids, putatively involved in the formation of caries were quantified using mass spectrometry (MS)-based techniques, such as (stable isotope dilution analysis)-ultra-performance liquid chromatography-tandem MS and gas chromatography/electron ionisation-MS. Pellicle and corresponding saliva samples were collected from caries-active, caries-free and caries-rehabilitated 4- to 6-year-old children. The most abundant analytes in pellicle were acetic acid (1.2–10.5 nmol/cm 2), propionic acid (0.1–8.5 nmol/cm 2), glycine (0.7–3.5 nmol/cm 2), serine (0.08–2.3 nmol/cm 2), galactose (galactose + mannose; 0.035–0.078 nmol/cm 2), lactose (0.002–0.086 nmol/cm 2), glucose (0.018–0.953 nmol/cm 2), palmitic acid (0.26–2.03 nmol/cm 2), and stearic acid (0.34–1.81 nmol/cm 2). Significant differences depending on caries activity were detected neither in saliva nor in the corresponding pellicle samples.

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          An isotope-labeled chemical derivatization method for the quantitation of short-chain fatty acids in human feces by liquid chromatography-tandem mass spectrometry.

          Short-chain fatty acids (SCFAs) are produced by anaerobic gut microbiota in the large bowel. Qualitative and quantitative measurements of SCFAs in the intestinal tract and the fecal samples are important to understand the complex interplay between diet, gut microbiota and host metabolism homeostasis. To develop a new LC-MS/MS method for sensitive and reliable analysis of SCFAs in human fecal samples, 3-nitrophenylhydrazine (3NPH) was employed for pre-analytical derivatization to convert ten C2-C6 SCFAs to their 3-nitrophenylhydrazones under a single set of optimized reaction conditions and without the need of reaction quenching. The derivatives showed excellent in-solution chemical stability. They were separated on a reversed-phase C18 column and quantitated by negative-ion electrospray ionization - multiple-reaction monitoring (MRM)/MS. To achieve accurate quantitation, the stable isotope-labeled versions of the derivatives were synthesized in a single reaction vessel from (13)C6-3NPH, and were used as internal standard to compensate for the matrix effects in ESI. Method validation showed on-column limits of detection and quantitation over the range from low to high femtomoles for the ten SCFAs, and the intra-day and inter-day precision for determination of nine of the ten SCFAs in human fecal samples was ≤8.8% (n=6). The quantitation accuracy ranged from 93.1% to 108.4% (CVs≤4.6%, n=6). This method was used to determine the SCFA concentrations and compositions in six human fecal samples. One of the six samples, which was collected from a clinically diagnosed type 2 diabetes patient showed a significantly high molar ratio of branch-chain SCFAs to straight-chain SCFAs than the others. In summary, this work provides a new LC-MS/MS method for precise and accurate quantitation of SCFAs in human feces.
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            Early Childhood Caries: Prevalence, Risk Factors, and Prevention

            Early childhood caries (ECC) is major oral health problem, mainly in socially disadvantaged populations. ECC affects infants and preschool children worldwide. The prevalence of ECC differs according to the group examined, and a prevalence of up to 85% has been reported for disadvantaged groups. ECC is the presence of one or more decayed, missing, or filled primary teeth in children aged 71 months (5 years) or younger. It begins with white-spot lesions in the upper primary incisors along the margin of the gingiva. If the disease continues, caries can progress, leading to complete destruction of the crown. The main risk factors in the development of ECC can be categorized as microbiological, dietary, and environmental risk factors. Even though it is largely a preventable condition, ECC remains one of the most common childhood diseases. The major contributing factors for the for the high prevalence of ECC are improper feeding practices, familial socioeconomic background, lack of parental education, and lack of access to dental care. Oral health plays an important role in children to maintain the oral functions and is required for eating, speech development, and a positive self-image. The review will focus on the prevalence, risk factors, and preventive strategies and the management of ECC.
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              Dental plaque as a microbial biofilm.

              P D Marsh (2015)
              New technologies have provided novel insights into how dental plaque functions as a biofilm. Confocal microscopy has confirmed that plaque has an open architecture similar to other biofilms, with channels and voids. Gradients develop in areas of dense biomass over short distances in key parameters that influence microbial growth and distribution. Bacteria exhibit an altered pattern of gene expression either as a direct result of being on a surface or indirectly as a response to the local environmental heterogeneity within the biofilm. Bacteria communicate via small diffusible signalling molecules (e.g. competence-stimulating peptide, CSP; autoinducer 2); CSP induces both genetic competence and acid tolerance in recipient sessile cells. Thus, rates of gene transfer increase in biofilm communities, and this is one of several mechanisms (others include: diffusion-reaction, neutralization/inactivation, slow growth rates, novel phenotype) that contribute to the increased antimicrobial resistance exhibited by bacteria in biofilms. Oral bacteria in plaque do not exist as independent entities but function as a co-ordinated, spatially organized and fully metabolically integrated microbial community, the properties of which are greater than the sum of the component species. A greater understanding of the significance of dental plaque as a mixed culture biofilm will lead to novel control strategies. Copyright 2004 S. Karger AG, Basel
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                Author and article information

                Contributors
                roman.lang@tum.de
                thomas.hofmann@tum.de
                Journal
                Sci Rep
                Sci Rep
                Scientific Reports
                Nature Publishing Group UK (London )
                2045-2322
                20 January 2020
                20 January 2020
                2020
                : 10
                : 697
                Affiliations
                [1 ]ISNI 0000000123222966, GRID grid.6936.a, Chair of Food Chemistry and Molecular Sensory Science, , Technical University of Munich, ; Lise-Meitner-Straße 34, D-85354 Freising, Germany
                [2 ]Bavarian Center for Biomolecular Mass Spectrometry, Gregor-Mendel-Straße 4, D-85354 Freising, Germany
                [3 ]Leibniz-Institute for Food Systems Biology at the Technical University of Munich, Lise-Meitner-Straße 34, D-85354 Freising, Germany
                [4 ]ISNI 0000 0001 2111 7257, GRID grid.4488.0, Policlinic of Operative and Pediatric Dentistry, Medical Faculty Carl Gustav Carus, , Technical University of Dresden, ; Fetscherstraße 74, D-01307 Dresden, Germany
                [5 ]ISNI 0000 0000 9130 6144, GRID grid.10211.33, Sustainable Chemistry and Material Resources, Institute of Sustainable and Environmental Chemistry, , Leuphana University of Lüneburg, ; Universitätsallee 1 C13, D-21335 Lüneburg, Germany
                [6 ]Clinic of Operative Dentistry and Periodontology, Saarland University, Universitätsklinikum des Saarlandes, D-66421 Homburg/Saar, Kirrberger Straße Germany
                Author information
                http://orcid.org/0000-0003-0610-7186
                http://orcid.org/0000-0001-8782-4456
                Article
                57531
                10.1038/s41598-020-57531-8
                6971297
                31959821
                2ac8960f-5f88-4068-993f-15c1893bffef
                © The Author(s) 2020

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 27 September 2019
                : 12 December 2019
                Funding
                Funded by: FundRef https://doi.org/10.13039/501100001659, Deutsche Forschungsgemeinschaft (German Research Foundation);
                Award ID: DFGHA 5192/11-1
                Award ID: DFGHA 2718/17-1
                Award ID: DFGHA 2718/17-1
                Award ID: DFGHO 2116/15-1
                Award ID: DFGHA 5192/11-1
                Award ID: DFGHA 5192/11-1
                Award ID: DFGHA 2718/17-1
                Award ID: DFGHO 2116/15-1
                Award ID: DFGHA 5192/11-1
                Award ID: DFGHA 2718/17-1
                Award ID: DFGHO 2116/15-1
                Award ID: DFGHA 5192/11-1
                Award ID: DFGHA 2718/17-1
                Award ID: DFGHO 2116/15-1
                Award Recipient :
                Categories
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                Custom metadata
                © The Author(s) 2020

                Uncategorized
                dental caries,quality of life,paediatric dentistry
                Uncategorized
                dental caries, quality of life, paediatric dentistry

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