Development and characterization of an oral microbiome transplant among Australians for the treatment of dental caries and periodontal disease: A study protocol

Background Oral microbiome transplantation (OMT) is a novel concept of introducing health-associated oral microbiota into the oral cavity of a diseased patient. The premise is to reverse the state of oral dysbiosis, and restore the ecological balance to maintain a stable homeostasis with the host immune system. This study will assess the effectiveness, feasibility, and safety of OMT using an interdisciplinary approach. Methods/Design To find donors suitable for microbial transplantation, supragingival plaque samples will be collected from 600 healthy participants. Each sample (200μL) will subsequently be examined in two ways: 1) 100μL of the sample will undergo high-throughput 16S rRNA gene amplicon sequencing and shotgun sequencing to identify the composition and characterisation of a healthy supragingival microbiome, 2) the remaining 100μL of the plaque sample will be mixed with 25% artificial saliva medium and inoculated into a specialised in-vitro flow cell model containing a hydroxyapatite disk. To obtain sufficient donor plaque, the samples would be grown for 14 days and further analysed microscopically and sequenced to examine and confirm the growth and survival of the microbiota. Samples with the healthiest microbiota would then be incorporated in a hydrogel delivery vehicle to enable transplantation of the donor oral microbiota. The third step would be to test the effectiveness of OMT in caries and periodontitis animal models for efficacy and safety for the treatment of oral diseases. Discussion If OMTs are found to be successful, it can form a new treatment method for common oral diseases such as dental caries and periodontitis. OMTs may have the potential to modulate the oral microbiota and shift the ecological imbalances to a healthier state.

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Oral microbiota and dental caries data from monozygotic and dizygotic twin children

Scientific Data ◽

10.1038/s41597-020-00691-z ◽

2020 ◽

Vol 7 (1) ◽

Author(s):

Yelda Kasimoglu ◽

Mine Koruyucu ◽

Sinem Birant ◽

Ilker Karacan ◽

Nursen Topcuoglu ◽

...

Keyword(s):

Dental Caries ◽

16S Rrna ◽

Clinical Investigation ◽

Monozygotic Twins ◽

Amplicon Sequencing ◽

Oral Microbiome ◽

Rrna Gene ◽

Oral Microbiota ◽

Microbiome Profile ◽

Twin Children

Abstract There are recent studies which aimed to detect the inheritance on the etiology of dental caries exploring oral composition. We present data on the oral microbiota and its relation with dental caries and other factors in monozygotic (MZ) and dizygotic (DZ) twin children. Following clinical investigation, DNA samples were collected and isolated from saliva of 198 patients (49 MZ and 50 DZ twins) with an average age of 9.7 ± 2.7 years. Salivary bacterial microbiota analysis was performed using high throughput amplicon sequencing method targeting V3-V4 region of the 16S rRNA gene. A total of 8,297,859 raw reads corresponding to 41,908 reads per sample were obtained on average. The QIIME2-deblur workflow was used for 16S rRNA amplicon analysis. Microbiome similarity analyses between twins (based on Bray-Curtis dissimilarity, weighted and unweighted Unifrac distances) showed that monozygotic twins share more bacterial microbial content compared to dizygotic twins. This is a large microbial community dataset of MZ and DZ twins with or without dental findings which can be further used for children oral microbiome profile explorations.

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Deep metagenomics examines the oral microbiome during dental caries, revealing novel taxa and co-occurrences with host molecules

10.1101/804443 ◽

2019 ◽

Cited By ~ 4

Author(s):

J.L. Baker ◽

J.T. Morton ◽

M. Dinis ◽

R. Alverez ◽

N.C. Tran ◽

...

Keyword(s):

Dental Caries ◽

Compositional Data ◽

Amplicon Sequencing ◽

Oral Microbiome ◽

Rrna Gene ◽

Host Immune System ◽

Metagenomic Sequencing ◽

Microbiome Composition ◽

Immunological Markers ◽

Large Populations

AbstractDental caries is the most common chronic infectious disease globally. The microbial communities associated with caries have mainly been examined using relatively low-resolution 16S rRNA gene amplicon sequencing and/or using downstream analyses that are unsound for the compositional nature of the data provided by sequencing. Additionally, the relationship between caries, oral microbiome composition, and host immunological markers has not been explored. In this study, the oral microbiome and a panel of 38 host markers was analyzed across the saliva from 23 children with dentin caries and 24 children with healthy dentition. Metagenomic sequencing, followed by investigation using tools designed to be robust for compositional data, illustrated that several Prevotella spp. were prevalent in caries, while Rothia spp. were associated with the health. The contributional diversity (extent to which multiple taxa contribute to each pathway) of functional pathways present in the oral microbiome was decreased in the caries group. This decrease was especially noticeable in several pathways known to impede caries pathogenesis, including arginine and branched-chain amino acid biosynthesis. 10 host immunological markers were found to be significantly elevated in the saliva of the caries group, and microbe-metabolite co-occurrence analysis provided an atlas of relationships contributing to the bi-directional influence between the oral microbiome and the host immune system. Finally, 527 metagenome-assembled genomes were obtained from the metagenomics data, representing 151 species. 23 taxa were novel genera/species and a further 20 taxa were novel species. This study thus serves as a model analysis pipeline that will tremendously expand our knowledge of the oral microbiome and its relationship to dental caries once applied to large populations.

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Tetramic Acids Mutanocyclin and Reutericyclin A, Produced by Streptococcus mutans Strain B04Sm5 Modulate the Ecology of an in vitro Oral Biofilm

Frontiers in Oral Health ◽

10.3389/froh.2021.796140 ◽

2022 ◽

Vol 2 ◽

Author(s):

Carla Uranga ◽

Karen E. Nelson ◽

Anna Edlund ◽

Jonathon L. Baker

Keyword(s):

Streptococcus Mutans ◽

Specific Effect ◽

16S Rrna Gene Sequencing ◽

Oral Microbiome ◽

Public Health Issue ◽

Rrna Gene ◽

Oral Microbiota ◽

Oral Biofilm ◽

Tetramic Acids

The human oral microbiome consists of diverse microbes actively communicating and interacting through a variety of biochemical mechanisms. Dental caries is a major public health issue caused by fermentable carbohydrate consumption that leads to dysbiosis of the oral microbiome. Streptococcus mutans is a known major contributor to caries pathogenesis, due to its exceptional ability to form biofilms in the presence of sucrose, as well as to its acidophilic lifestyle. S. mutans can also kill competing bacteria, which are typically health associated, through the production of bacteriocins and other small molecules. A subset of S. mutans strains encode the muc biosynthetic gene cluster (BGC), which was recently shown to produce the tetramic acids, mutanocyclin and reutericyclins A, B, and C. Reutericyclin A displayed strong antimicrobial activity and mutanocyclin appeared to be anti-inflammatory; however the effect of these compounds, and the carriage of muc by S. mutans, on the ecology of the oral microbiota is not known, and was examined here using a previously developed in vitro biofilm model derived from human saliva. While reutericyclin significantly inhibited in vitro biofilm formation and acid production at sub-nanomolar concentrations, mutanocyclin did not present any activity until the high micromolar range. 16S rRNA gene sequencing revealed that reutericyclin drastically altered the biofilm community composition, while mutanocyclin showed a more specific effect, reducing the relative abundance of cariogenic Limosilactobacillus fermentum. Mutanocyclin or reutericyclin produced by the S. mutans strains amended to the community did not appear to affect the community in the same way as the purified compounds, although the results were somewhat confounded by the differing growth rates of the S. mutans strains. Regardless of the strain added, the addition of S. mutans to the in vitro community significantly increased the abundance of S. mutans and Veillonella infantium, only. Overall, this study illustrates that reutericyclin A and mutanocyclin do impact the ecology of a complex in vitro oral biofilm; however, further research is needed to determine the extent to which the production of these compounds affects the virulence of S. mutans.

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Oral Microbiota Identifies Patients in Early Onset Rheumatoid Arthritis

Microorganisms ◽

10.3390/microorganisms9081657 ◽

2021 ◽

Vol 9 (8) ◽

pp. 1657

Author(s):

Anders Esberg ◽

Linda Johansson ◽

Ingegerd Johansson ◽

Solbritt Rantapää Dahlqvist

Keyword(s):

Rheumatoid Arthritis ◽

Early Onset ◽

Amplicon Sequencing ◽

Oral Bacteria ◽

Oral Microbiome ◽

Rrna Gene ◽

Oral Microbiota ◽

Disease Manifestation ◽

Periodontal Status ◽

Quality Register

Rheumatoid arthritis (RA) is the most common autoimmune inflammatory disease, and single periodontitis-associated bacteria have been suggested in disease manifestation. Here, the oral microbiota was characterized in relation to the early onset of RA (eRA) taking periodontal status into consideration. 16S rRNA gene amplicon sequencing of saliva bacterial DNA from 61 eRA patients without disease-modifying anti-rheumatic drugs and 59 matched controls was performed. Taxonomic classification at 98.5% was conducted against the Human Oral Microbiome Database, microbiota functions were predicted using PICRUSt, and periodontal status linked from the Swedish quality register for clinically assessed caries and periodontitis. The participants were classified into three distinct microbiota-based cluster groups with cluster allocation differences by eRA status. Independently of periodontal status, eRA patients had enriched levels of Prevotella pleuritidis, Treponema denticola, Porphyromonas endodontalis and Filifactor alocis species and in the Porphyromonas and Fusobacterium genera and functions linked to ornithine metabolism, glucosylceramidase, beta-lactamase resistance, biphenyl degradation, fatty acid metabolism and 17-beta-estradiol-17-dehydrogenase metabolism. The results support a deviating oral microbiota composition already in eRA patients compared with healthy controls and highlight a panel of oral bacteria that may be useful in eRA risk assessment in both periodontally healthy and diseased persons.

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OTUs clustering should be avoided for defining oral microbiome

10.1101/2021.08.09.455616 ◽

2021 ◽

Author(s):

Alba Regueira-Iglesias ◽

Lara Vazquez-Gonzalez ◽

Carlos Balsa-Castro ◽

Triana Blanco-Pintos ◽

Victor Manuel Arce ◽

...

Keyword(s):

Microbial Diversity ◽

Operational Taxonomic Unit ◽

Oral Bacteria ◽

Oral Microbiome ◽

Rrna Gene ◽

Oral Microbiota ◽

High Coverage ◽

Archaeal Species ◽

Health And Disease ◽

Similarity Relationships

This in silico investigation aimed to: 1) evaluate a set of primer pairs with high coverage, including those most commonly used in the literature, to find the different oral species with 16S rRNA gene amplicon similarity/identity (ASI) values ≥97%; and 2) identify oral species that may be erroneously clustered in the same operational taxonomic unit (OTU) and ascertain whether they belong to distinct genera or other higher taxonomic ranks. Thirty-nine primer pairs were employed to obtain amplicon sequence variants (ASVs) from the complete genomes of 186 bacterial and 135 archaeal species. For each primer, ASVs without mismatches were aligned using BLASTN and their similarity values were obtained. Finally, we selected ASVs from different species with an ASI value ≥97% that were covered 100% by the query sequences. For each primer, the percentage of species-level coverage with no ASI≥97% (SC-NASI≥97%) was calculated. Based on the SC-NASI≥97% values, the best primer pairs were OP_F053-KP_R020 for bacteria (65.05%), KP_F018-KP_R002 for archaea (51.11%), and OP_F114-KP_R031 for bacteria and archaea together (52.02%). Eighty percent of the oral-bacteria and oral-archaea species shared an ASI≥97% with at least one other taxa, including Campylobacter, Rothia, Streptococcus, and Tannerella, which played conflicting roles in the oral microbiota. Moreover, around a quarter and a third of these two-by-two similarity relationships were between species from different bacteria and archaea genera, respectively. Furthermore, even taxa from distinct families, orders, and classes could be grouped in the same cluster. Consequently, irrespective of the primer pair used, OTUs constructed with a 97% similarity provide an inaccurate description of oral-bacterial and oral-archaeal species, greatly affecting microbial diversity parameters. As a result, clustering by OTUs impacts the credibility of the associations between some oral species and certain health and disease conditions. This limits significantly the comparability of the microbial diversity findings reported in oral microbiome literature.

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Impact of Oropharyngeal Administration of Colostrum in Preterm Newborns’ Oral Microbiome

Nutrients ◽

10.3390/nu13124224 ◽

2021 ◽

Vol 13 (12) ◽

pp. 4224

Author(s):

Ramon V. Cortez ◽

Andrea Fernandes ◽

Luiz Gustavo Sparvoli ◽

Marina Padilha ◽

Rubens Feferbaum ◽

...

Keyword(s):

Preterm Infants ◽

16S Rrna Gene Sequencing ◽

Oral Microbiome ◽

Rrna Gene ◽

Oral Microbiota ◽

Alpha And Beta Diversity ◽

Longitudinal Observational Study ◽

Key Factor ◽

Rrna Gene Sequencing ◽

Preterm Newborns

The initial colonization of the human microbiota is of paramount importance. In this context, the oropharyngeal administration of colostrum is a safe, viable, and well-tolerated practice even by the smallest preterm infants. Therefore, this study evaluated the effects of oropharyngeal administration of colostrum on the establishment of preterm infants’ oral microbiota. A longitudinal observational study was carried out with 20 premature neonates, divided into two groups: one receiving the protocol (Oropharyngeal Administration of Colostrum; OAC) and the other one receiving Standard Caare (SC). Saliva samples were collected from the newborns weekly during the study period (from the day of birth until the 21st day of life) for analysis of oral microbiota through 16S rRNA gene sequencing. We observed that the colonization of the oral microbiota of preterm newborns preseanted a higher relative abundance of Staphylococcus on the 7th day of life, mainly in the OAC group. Additionally, an increased abundance of Bifidobacterium and Bacteroides was observed in the OAC group at the first week of life. Regarding alpha and beta diversity, time was a key factor in the oral modulation of both groups, showing how dynamic this environment is in early life.

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Human oral microbiome and prospective risk for pancreatic cancer: a population-based nested case-control study

Gut ◽

10.1136/gutjnl-2016-312580 ◽

2016 ◽

Vol 67 (1) ◽

pp. 120-127 ◽

Cited By ~ 177

Author(s):

Xiaozhou Fan ◽

Alexander V Alekseyenko ◽

Jing Wu ◽

Brandilyn A Peters ◽

Eric J Jacobs ◽

...

Keyword(s):

Pancreatic Cancer ◽

Case Control Study ◽

Case Control ◽

Oral Microbiome ◽

Rrna Gene ◽

Oral Microbiota ◽

Increased Risk ◽

Oral Pathogens ◽

Nested Case Control ◽

Control Study

ObjectiveA history of periodontal disease and the presence of circulating antibodies to selected oral pathogens have been associated with increased risk of pancreatic cancer; however, direct relationships of oral microbes with pancreatic cancer have not been evaluated in prospective studies. We examine the relationship of oral microbiota with subsequent risk of pancreatic cancer in a large nested case–control study.DesignWe selected 361 incident adenocarcinoma of pancreas and 371 matched controls from two prospective cohort studies, the American Cancer Society Cancer Prevention Study II and the National Cancer Institute Prostate, Lung, Colorectal and Ovarian Cancer Screening Trial. From pre-diagnostic oral wash samples, we characterised the composition of the oral microbiota using bacterial 16S ribosomal RNA (16S rRNA) gene sequencing. The associations between oral microbiota and risk of pancreatic cancer, controlling for the random effect of cohorts and other covariates, were examined using traditional and L1-penalised least absolute shrinkage and selection operator logistic regression.ResultsCarriage of oral pathogens, Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans, were associated with higher risk of pancreatic cancer (adjusted OR for presence vs absence=1.60 and 95% CI 1.15 to 2.22; OR=2.20 and 95% CI 1.16 to 4.18, respectively). Phylum Fusobacteria and its genus Leptotrichia were associated with decreased pancreatic cancer risk (OR per per cent increase of relative abundance=0.94 and 95% CI 0.89 to 0.99; OR=0.87 and 95% CI 0.79 to 0.95, respectively). Risks related to these phylotypes remained after exclusion of cases that developed within 2 years of sample collection, reducing the likelihood of reverse causation in this prospective study.ConclusionsThis study provides supportive evidence that oral microbiota may play a role in the aetiology of pancreatic cancer.

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Multifunctional artificial saliva preparation – challenges and needs

Postępy Polskiej Medycyny i Farmacji ◽

10.5604/01.3001.0014.9199 ◽

2021 ◽

Vol 8 ◽

pp. 37-46

Author(s):

Katarzyna Niemirowicz-Laskowska ◽

Joanna Mystkowska ◽

Dawid Łysik ◽

Sylwia Chmielewska ◽

Łukasz Suprewicz ◽

...

Keyword(s):

Oral Cavity ◽

Artificial Saliva ◽

Salivary Secretion ◽

Biological Properties ◽

Pharmacological Therapy ◽

Oral Microbiome ◽

Dry Mouth ◽

Rheological Parameters ◽

Saliva Substitutes

Saliva plays a crucial role in maintaining homeostasis not only within the oral cavity but also in further sections of the gastrointestinal tract. Pleiotropic properties of saliva include participation in the digestion of carbohydrates, cleansing and moisturizing the oral cavity, and maintaining the composition of the oral microbiome. The result of impaired function of the salivary gland is reduced salivation – hyposalivation, leading to dry mouth – xerostomia. It is established that numerous physiological factors (age, sex, weight change) and pathological factors (polytherapy, head and neck cancer, coexisting diseases such as diabetes, depression, cardiovascular diseases) lead to the reduction in saliva secretion, and in effect, causing a dry mouth. Treatment of salivary secretion disorders involves pharmacological therapy (including hormone therapy) or replacement therapy which based on the use of saliva substitutes. In the case of disturbances in the secretion of natural saliva, the application of the artificial saliva preparations should support the chewing processes, moisturize the oral cavity, and fulfill the biological functions of saliva. However, to date, on the pharmaceutical market, there are no saliva substitutes that meet the biological criteria and maintaining favorable physicochemical properties and rheological parameters. Taking into account the problems of the patients which are burden by impaired salivary secretion, the aim of our research was to attempt to develop an artificial saliva preparation that reflecting as much as possible the properties of natural saliva, both in terms of mechanical and biological properties. As part of the research, the chemical composition was developed and a detailed study of the physicochemical and rheological parameters of artificial saliva preparations containing mucins as well as their microbiological and biocompatibility assessment, at in vitro level were carried out.

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Analysis of the Effects of Dietary Pattern on the Oral Microbiome of Elite Endurance Athletes

Nutrients ◽

10.3390/nu11030614 ◽

2019 ◽

Vol 11 (3) ◽

pp. 614 ◽

Cited By ~ 17

Author(s):

Nida Murtaza ◽

Louise Burke ◽

Nicole Vlahovich ◽

Bronwen Charlesson ◽

Hayley O’Neill ◽

...

Keyword(s):

Discriminant Analysis ◽

Relative Abundance ◽

Distance Measure ◽

Oral Microbiome ◽

Rrna Gene ◽

Oral Microbiota ◽

Unifrac Distance ◽

Linear Discriminant ◽

Principal Coordinates ◽

Low Carbohydrate

Although the oral microbiota is known to play a crucial role in human health, there are few studies of diet x oral microbiota interactions, and none in elite athletes who may manipulate their intakes of macronutrients to achieve different metabolic adaptations in pursuit of optimal endurance performance. The aim of this study was to investigate the shifts in the oral microbiome of elite male endurance race walkers from Europe, Asia, the Americas and Australia, in response to one of three dietary patterns often used by athletes during a period of intensified training: a High Carbohydrate (HCHO; n = 9; with 60% energy intake from carbohydrates; ~8.5 g kg−1 day−1 carbohydrate, ~2.1 g kg−1 day−1 protein, 1.2 g kg−1 day−1 fat) diet, a Periodised Carbohydrate (PCHO; n = 10; same macronutrient composition as HCHO, but the intake of carbohydrates is different across the day and throughout the week to support training sessions with high or low carbohydrate availability) diet or a ketogenic Low Carbohydrate High Fat (LCHF; n = 10; 0.5 g kg−1 day−1 carbohydrate; 78% energy as fat; 2.1 g kg−1 day−1 protein) diet. Saliva samples were collected both before (Baseline; BL) and after the three-week period (Post treatment; PT) and the oral microbiota profiles for each athlete were produced by 16S rRNA gene amplicon sequencing. Principal coordinates analysis of the oral microbiota profiles based on the weighted UniFrac distance measure did not reveal any specific clustering with respect to diet or athlete ethnic origin, either at baseline (BL) or following the diet-training period. However, discriminant analyses of the oral microbiota profiles by Linear Discriminant Analysis (LDA) Effect Size (LEfSe) and sparse Partial Least Squares Discriminant Analysis (sPLS-DA) did reveal changes in the relative abundance of specific bacterial taxa, and, particularly, when comparing the microbiota profiles following consumption of the carbohydrate-based diets with the LCHF diet. These analyses showed that following consumption of the LCHF diet the relative abundances of Haemophilus, Neisseria and Prevotella spp. were decreased, and the relative abundance of Streptococcus spp. was increased. Such findings suggest that diet, and, in particular, the LCHF diet can induce changes in the oral microbiota of elite endurance walkers.

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Comparison of Oral Microbiota Collected Using Multiple Methods and Recommendations for New Epidemiologic Studies

mSystems ◽

10.1128/msystems.00156-20 ◽

2020 ◽

Vol 5 (4) ◽

Cited By ~ 2

Author(s):

Yukiko Yano ◽

Xing Hua ◽

Yunhu Wan ◽

Shalabh Suman ◽

Bin Zhu ◽

...

Keyword(s):

Beta Diversity ◽

Epidemiologic Studies ◽

Oral Microbiome ◽

Rrna Gene ◽

Oral Microbiota ◽

Collection Method ◽

Two Samples ◽

Relative Abundances ◽

Collection Methods ◽

Diversity Metrics

ABSTRACT Epidemiologic studies use various biosample collection methods to study associations between human oral microbiota and health outcomes. However, the agreement between the different methods is unclear. We compared a commercially available OMNIgene ORAL kit to three alternative collection methods: Saccomanno’s fixative, Scope mouthwash, and nonethanol mouthwash. Oral samples were collected from 40 individuals over 4 visits. Two samples were collected from each subject per visit: one with OMNIgene and one with an alternative method. DNA was extracted using the DSP DNA Virus Pathogen kit, and the V4 region of the 16S rRNA gene was PCR amplified and sequenced using MiSeq. Oral collection methods were compared based on alpha and beta diversity metrics and phylum- and genus-level relative abundances. All alpha diversity metrics were significantly lower for Saccomanno’s fixative than for OMNIgene (P < 0.001), whereas the two mouthwashes were more similar to OMNIgene. Principal-coordinate analysis (PCoA) using the Bray-Curtis and weighted UniFrac beta diversity matrices showed large differences in the microbial compositions of samples collected with Saccomanno’s compared to those with OMNIgene and the mouthwashes. Clustering by collection method was not observed in unweighted UniFrac PCoA plots, suggesting differences in relative abundances but not specific taxa detected by the collection methods. Relative abundances of most taxa were significantly different between OMNIgene and the other methods at each taxonomic level, with Saccomanno’s showing the least agreement with OMNIgene. There were clear differences in oral microbial communities between the four oral collection methods, particularly for Saccomanno’s fixative. IMPORTANCE We compared four different oral collection methods for studying the human oral microbiome: an OMNIgene ORAL kit, Scope mouthwash, nonethanol mouthwash, and Saccomanno’s fixative. Our study shows that the type of the collection method can have a large impact on the results of an oral microbiome analysis. We recommend that one consistent oral collection method should be used for all oral microbiome comparisons. While Scope and nonethanol mouthwashes are less expensive and provide results similar to those with OMNIgene, Saccomanno’s fixative may be unfavorable due to the microbial differences detected in this study. Our results will help guide the design of future oral microbiome studies.

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