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1.
Biomolecules ; 11(8)2021 08 06.
Artigo em Inglês | MEDLINE | ID: mdl-34439831

RESUMO

Wound healing is an essential process to restore tissue integrity after trauma. Large skin wounds such as burns often heal with hypertrophic scarring and contractures, resulting in disfigurements and reduced joint mobility. Such adverse healing outcomes are less common in the oral mucosa, which generally heals faster compared to skin. Several studies have identified differences between oral and skin wound healing. Most of these studies however focus only on a single stage of wound healing or a single cell type. The aim of this review is to provide an extensive overview of wound healing in skin versus oral mucosa during all stages of wound healing and including all cell types and molecules involved in the process and also taking into account environmental specific factors such as exposure to saliva and the microbiome. Next to intrinsic properties of resident cells and differential expression of cytokines and growth factors, multiple external factors have been identified that contribute to oral wound healing. It can be concluded that faster wound closure, the presence of saliva, a more rapid immune response, and increased extracellular matrix remodeling all contribute to the superior wound healing and reduced scar formation in oral mucosa, compared to skin.


Assuntos
Matriz Extracelular/imunologia , Microbiota/imunologia , Mucosa Bucal/lesões , Pele/lesões , Cicatrização/imunologia , Animais , Citocinas/genética , Citocinas/imunologia , Matriz Extracelular/química , Fibroblastos/imunologia , Fibroblastos/microbiologia , Regulação da Expressão Gênica , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/genética , Peptídeos e Proteínas de Sinalização Intercelular/imunologia , Queratinócitos/imunologia , Queratinócitos/microbiologia , Macrófagos/imunologia , Macrófagos/microbiologia , Mucosa Bucal/imunologia , Mucosa Bucal/microbiologia , Mucosa Bucal/patologia , Neutrófilos/imunologia , Neutrófilos/microbiologia , Especificidade de Órgãos , Saliva/imunologia , Saliva/microbiologia , Transdução de Sinais , Pele/imunologia , Pele/microbiologia , Pele/patologia
2.
Front Immunol ; 12: 626217, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34276643

RESUMO

Alterations in the microbiome of the gut and oral cavity are involved in the etiopathogenesis of systemic lupus erythematosus (SLE). We aimed to assess whether both microbiome compositions in feces and saliva were specific in patients with SLE. A total of 35 patients with SLE, as well as sex- and age-matched asymptomatic subjects as healthy control (HC) group were recruited. Fecal swabs and saliva samples were collected from the participants. 16S ribosomal RNA gene sequencing was performed on the samples. Compared with the HC group, reduced bacterial richness and diversity were detected in the feces of patients with SLE, and increased bacterial diversity in their saliva. Both feces and saliva samples explained the cohort variation. The feces were characterized by enrichment of Lactobacillus, and depletion of an unclassified bacterium in the Ruminococcaceae family and Bifidobacterium. Lack of Bifidobacterium was observed in patients with arthritis. Akkermansia and Ruminococcus negatively correlated with the serum levels of C3. In saliva, Veillonella, Streptococcus, and Prevotella were dominant, and Bacteroides was negatively associated with disease activity. These findings can assist us to comprehensively understand the bacterial profiles of different body niches in SLE patients.


Assuntos
Bactérias/genética , Microbioma Gastrointestinal/genética , Lúpus Eritematoso Sistêmico/microbiologia , Microbiota/genética , Saliva/microbiologia , Adulto , Idoso , Bactérias/classificação , Estudos de Coortes , Fezes/microbiologia , Feminino , Microbioma Gastrointestinal/fisiologia , Variação Genética , Humanos , Masculino , Microbiota/fisiologia , Pessoa de Meia-Idade , Boca/microbiologia , Adulto Jovem
3.
Forensic Sci Int ; 325: 110886, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34192646

RESUMO

Sexual assault offences are one of the most serious crimes committed against a person, typically rank only second to homicide, and represent one of the major challenges in forensic sciences. In some cases of sexual assault, there may be more than one suspect and the analysis of the biological evidence with currently available methods such as human DNA analysis may not yield results. In this study using the designed experimental model (with different experimental scenarios that can be designed), it was aimed to investigate the effectiveness of the microbiome profile for the identification of the offender by comparing the microbiome structures of the suspects' saliva samples with the mixed samples on the victim (saliva transmitted on breast skin) within the first 48 h after a sexual assault. For this purpose, a total of 44 samples was collected from four healthy females and four healthy males aged 20-50 years. Microbiome profiles of 44 samples in four groups containing saliva, breast skin and mixed samples were determined with the IIlumina HiSeq platform. Differentiation between samples were calculated by beta-diversity analysis methods by using QIIME software (v1.80). To compare the differentiation among samples and groups, unweighted UniFrac distance values were applied. Eight dominant microbial genera accounted for 86.15% of the total bacterial population in male saliva samples and were composed of Fusobacterium, Haemophilus, Neisseria, Porphyromonas, Prevotella, Rothia, Streptococcus and Veillonella. These genera constituted 76.72% of the bacterial population in mixed samples, whereas they constituted 34.40% of the bacterial population in the breast skin samples. Results of this study show that bacterial DNA in saliva can be recovered from saliva transmitted breast skin within at least 48 h. In conclusion, it has been found that examination of the microbiota of the saliva transmitted to breast skin of a sexual assault victim as a forensic tool may have the potential to determine the offender of the incident among the suspects or to reduce the number of suspects. Supporting the results of this study with further studies using parameters such as different case models, different body regions, larger time periods and a higher number of participants will be beneficial to draw accurate conclusion of the judicial case.


Assuntos
Mama , Saliva/microbiologia , Delitos Sexuais , Pele/microbiologia , Adulto , DNA Bacteriano/genética , Feminino , Medicina Legal/métodos , Humanos , Masculino , Microbiota , Pessoa de Meia-Idade , RNA Ribossômico 16S , Análise de Sequência de DNA , Adulto Jovem
4.
Forensic Sci Int Genet ; 53: 102537, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-34090061

RESUMO

Current body fluid identification methods do not reveal any information about the time since deposition (TsD) of biological traces, even though determining the age of traces could be crucial for the investigative process. To determine the utility of microbial RNA markers for TsD estimation, we examined RNA sequencing data from five forensically relevant body fluids (blood, menstrual blood, saliva, semen, and vaginal secretion) over seven time points, ranging from fresh to 1.5 years. One set of samples was stored indoors while another was exposed to outdoor conditions. In outdoor samples, we observed a consistent compositional shift, occurring after 4 weeks: this shift was characterized by an overall increase in non-human eukaryotic RNA and an overall decrease in prokaryotic RNA. In depth analyses showed a high fraction of tree, grass and fungal signatures, which are characteristic for the environment the samples were exposed to. When examining the prokaryotic fraction in more detail, three bacterial phyla were found to exhibit the largest changes in abundance, namely Actinobacteria, Proteobacteria and Firmicutes. More detailed analyses at the order level were done using a Lasso regression analysis to find a predictive subset of bacterial taxa. We found 26 bacterial orders to be indicative of sample age. Indoor samples did not reveal such a clear compositional change at the domain level: eukaryotic and prokaryotic abundance remained relatively stable across the assessed time period. Nonetheless, a Lasso regression analysis identified 32 bacterial orders exhibiting clear changes over time, enabling the prediction of TsD. For both indoor and outdoor samples, a larger number (around 60%) of the bacterial orders identified as indicative of TsD are part of the Actinobacteria, Proteobacteria and Firmicutes. In summary, we found that the observed changes across time are not primarily due to changes associated with body fluid specific bacteria but mostly due to accumulation of bacteria from the environment. Orders of these environmental bacteria could be evaluated for TsD prediction, considering the location and environment of the crime scene. However, further studies are needed to verify these findings, determine the applicability across samples, replicates, donors, and other variables, and also to further assess the effect of different seasons and locations on the samples.


Assuntos
Sangue/microbiologia , Muco do Colo Uterino/microbiologia , RNA Bacteriano/análise , Saliva/microbiologia , Sêmen/microbiologia , Análise de Sequência de RNA , Crime , Impressões Digitais de DNA , Exposição Ambiental , Feminino , Marcadores Genéticos , Humanos , Masculino , Menstruação , Repetições de Microssatélites , Fatores de Tempo
5.
J Vet Diagn Invest ; 33(4): 655-663, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34075814

RESUMO

The use of oral fluid (OF) to detect zoonotic pathogens in pigs has been only scarcely assessed. We evaluated OF as a potential specimen for detection by culture of methicillin-resistant Staphylococcus aureus (MRSA) and Yersinia enterocolitica, and the detection of antibodies against Salmonella spp. and hepatitis E virus (HEV) using commercial ELISAs. Samples from 33 pig farms were collected at the beginning and end of the fattening period. Results of the OF samples were compared with the results of serum samples and nasal swabs from individual pigs and pen floor fecal samples, using the Cohen kappa (κ) and the McNemar test. For Salmonella spp. antibodies, OF samples were negative, although the corresponding serum samples were positive. The detection of HEV antibodies in sera and OF had agreement at the first sampling, and poor and significant agreement at the second sampling (κ = 0.185, McNemar p = 0.238; κ = 0.088, McNemar p < 0.001). At both sampling times, the detection of MRSA in nasal swabs and OF showed agreement (κ = 0.466, McNemar p = 0.077; κ = 0.603, McNemar p = 1); agreement was seen for the detection of Y. enterocolitica in fecal and OF samples (κ = 0.012, McNemar p = 0.868; κ = 0.082, McNemar p = 0.061, respectively). According to the McNemar test, the use of pen-based OFs is more feasible for the detection of MRSA and Y. enterocolitica by culture than is detection of antibodies by commercial ELISA.


Assuntos
Hepatite E/veterinária , Saliva/microbiologia , Salmonelose Animal/epidemiologia , Infecções Estafilocócicas/veterinária , Doenças dos Suínos/epidemiologia , Yersiniose/veterinária , Animais , Hepatite E/diagnóstico , Hepatite E/epidemiologia , Hepatite E/microbiologia , Vírus da Hepatite E/isolamento & purificação , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Prevalência , Salmonella/isolamento & purificação , Salmonelose Animal/diagnóstico , Salmonelose Animal/microbiologia , Estudos Soroepidemiológicos , Infecções Estafilocócicas/diagnóstico , Infecções Estafilocócicas/epidemiologia , Infecções Estafilocócicas/microbiologia , Sus scrofa , Suínos , Doenças dos Suínos/diagnóstico , Doenças dos Suínos/microbiologia , Suíça/epidemiologia , Yersiniose/diagnóstico , Yersiniose/epidemiologia , Yersiniose/microbiologia , Yersinia enterocolitica/isolamento & purificação
6.
Microbes Environ ; 36(2)2021.
Artigo em Inglês | MEDLINE | ID: mdl-34078780

RESUMO

IgA nephropathy is one of the leading causes of chronic kidney disease in Japan. Since the origin and mechanisms by which IgA nephropathy develops currently remain unclear, a confirmed disease diagnosis is currently only possible by highly invasive renal biopsy. With the background of the salivary microbiome as a rich source of biomarkers for systemic diseases, we herein primarily aimed to investigate the salivary microbiome as a tool for the non-invasive diagnosis of IgA nephropathy. In a comparison of salivary microbiome profiles using 16S rRNA amplicon sequencing, significant differences were observed in microbial diversity and richness between IgA nephropathy patients and healthy controls. Furthermore, recent studies reported that patients with IgA nephropathy are more likely to develop inflammatory bowel diseases and that chronic inflammation of the tonsils triggered the recurrence of IgA nephropathy. Therefore, we compared the salivary microbiome of IgA nephropathy patients with chronic tonsillitis and ulcerative colitis patients. By combining the genera selected by the random forest algorithm, we were able to distinguish IgA nephropathy from healthy controls with an area under the curve (AUC) of 0.90, from the ulcerative colitis group with AUC of 0.88, and from the chronic tonsillitis group with AUC of 0.70. Additionally, the genus Neisseria was common among the selected genera that facilitated the separation of the IgA nephropathy group from healthy controls and the chronic tonsillitis group. The present results indicate the potential of the salivary microbiome as a biomarker for the non-invasive diagnosis of IgA nephropathy.


Assuntos
Bactérias/isolamento & purificação , Disbiose/microbiologia , Glomerulonefrite por IGA/microbiologia , Microbiota , Saliva/microbiologia , Adulto , Área Sob a Curva , Bactérias/classificação , Bactérias/genética , Estudos de Coortes , Feminino , Humanos , Japão , Masculino , Pessoa de Meia-Idade , Adulto Jovem
7.
Biochim Biophys Acta Proteins Proteom ; 1869(8): 140659, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-33839314

RESUMO

Saliva is a biofluid that maintains the health of oral tissues and the homeostasis of oral microbiota. Studies have demonstrated that Oral squamous cell carcinoma (OSCC) patients have different salivary microbiota than healthy individuals. However, the relationship between these microbial differences and clinicopathological outcomes is still far from conclusive. Herein, we investigate the capability of using metagenomic and metaproteomic saliva profiles to distinguish between Control (C), OSCC without active lesion (L0), and OSCC with active lesion (L1) patients. The results show that there are significantly distinct taxonomies and functional changes in L1 patients compared to C and L0 patients, suggesting compositional modulation of the oral microbiome, as the relative abundances of Centipeda, Veillonella, and Gemella suggested by metagenomics are correlated with tumor size, clinical stage, and active lesion. Metagenomics results also demonstrated that poor overall patient survival is associated with a higher relative abundance of Stenophotromonas, Staphylococcus, Centipeda, Selenomonas, Alloscordovia, and Acitenobacter. Finally, compositional and functional differences in the saliva content by metaproteomics analysis can distinguish healthy individuals from OSCC patients. In summary, our study suggests that oral microbiota and their protein abundance have potential diagnosis and prognosis value for oral cancer patients. Further studies are necessary to understand the role of uniquely detected metaproteins in the microbiota of healthy and OSCC patients as well as the crosstalk between saliva host proteins and the oral microbiome present in OSCC.


Assuntos
Saliva/microbiologia , Carcinoma de Células Escamosas de Cabeça e Pescoço/microbiologia , Adulto , Idoso , Estudos de Coortes , Feminino , Humanos , Masculino , Metagenômica/métodos , Microbiota/genética , Pessoa de Meia-Idade , Neoplasias Bucais/metabolismo , Neoplasias Bucais/microbiologia , Prognóstico , Proteômica/métodos , Carcinoma de Células Escamosas de Cabeça e Pescoço/metabolismo
8.
Molecules ; 26(7)2021 Mar 30.
Artigo em Inglês | MEDLINE | ID: mdl-33808410

RESUMO

Background: The study examined the oral microbiota, physiological and immunological changes in patients using thermoplastic retainers during three months of use. Methods: The study included several steps. Firstly, 10 swabs were collected from the buccal and palatal surfaces of the teeth of the patients, approximately 2 mL of saliva was collected from the same patients and 2 mL of saliva was collected from 10 healthy people to measure the pH and secretory IgA level. This was followed by the isolation and identfication of the bacterial isolates in the patient samples. Then, isolate susceptibility toward chlorhexidine (CHX) and their adhesion ability to thermoplastic retainer surfaces was measured. In addition to that the study estimated the numbers of Lactobacillus and Streptooccus mutans colonies during three months and finally, a comparsion of pH acidity and IgA level between the patients and healthy people was performed. The results showed the predominant bacteria during the three months were Lactobacillus spp. and Streptococcus spp. followed by different rates of other bacteria. Raoultella ornithinolytica showed more resistance to CHX while Lactobacillus spp. showed more sensitivity. Streptococcus mutans colony levels were higher than Lactobacillus spp. colonies during the three months, also S. mutans had the highest value in adherence to retainer thermoplastic. Finally, pH acidity showed a highly significant difference (p ≤ 0.05) in the third month, like IgA levels (p ≤ 0.05). Conclusions: According to the results obtained from the current study, the researchers noted that the thermoplastic retainers helped change the oral cavity environment.


Assuntos
Concentração de Íons de Hidrogênio , Imunoglobulina A Secretora/imunologia , Microbiota , Boca , Contenções Ortodônticas , Saliva , Humanos , Lactobacillus/isolamento & purificação , Boca/imunologia , Boca/microbiologia , Saliva/imunologia , Saliva/microbiologia , Streptococcus mutans/isolamento & purificação
9.
Molecules ; 26(5)2021 Mar 03.
Artigo em Inglês | MEDLINE | ID: mdl-33802588

RESUMO

In contrast to enamel, dentin surfaces have been rarely used as substrates for studies evaluating the effects of experimental rinsing solutions on oral biofilm formation. The aim of the present in situ study was to investigate the effects of tannic acid and chitosan on 48-h biofilm formation on dentin surfaces. Biofilm was formed intraorally on dentin specimens, while six subjects rinsed with experimental solutions containing tannic acid, chitosan and water as negative or chlorhexidine as positive control. After 48 h of biofilm formation, specimens were evaluated for biofilm coverage and for viability of bacteria by fluorescence and scanning electron microscopy. In addition, saliva samples were collected after rinsing and analyzed by fluorescence (five subjects) and transmission electron microscopy (two subjects) in order to investigate the antibacterial effect on bacteria in a planktonic state and to visualize effects of the rinsing agents on salivary proteins. After rinsing with water, dentin specimens were covered by a multiple-layered biofilm with predominantly vital bacteria. In contrast, chlorhexidine led to dentin surfaces covered only by few and avital bacteria. By rinsing with tannic acid both strong anti-adherent and antibacterial effects were observed, but the effects declined in a time-dependent manner. Transmission electron micrographs of salivary samples indicated that aggregation of proteins and bacteria might explain the antiadhesion effects of tannic acid. Chitosan showed antibacterial effects on bacteria in saliva, while biofilm viability was only slightly reduced and no effects on bacterial adherence on dentin were observed, despite proteins being aggregated in saliva after rinsing with chitosan. Tannic acid is a promising anti-biofilm agent even on dentin surfaces, while rinsing with chitosan could not sufficiently prevent biofilm formation on dentin.


Assuntos
Antibacterianos/farmacologia , Biofilmes/crescimento & desenvolvimento , Quitosana/farmacologia , Dentina/efeitos dos fármacos , Antissépticos Bucais/farmacologia , Saliva/efeitos dos fármacos , Taninos/farmacologia , Adulto , Aderência Bacteriana/efeitos dos fármacos , Biofilmes/efeitos dos fármacos , Quelantes/farmacologia , Estudos Cross-Over , Humanos , Saliva/microbiologia , Adulto Jovem
10.
PLoS One ; 16(4): e0251017, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33914825

RESUMO

BACKGROUND: Certain oral bacterial pathogens may play a role in oral carcinogenesis. We assessed the feasibility of conducting a population-based study in India to examine the distributions and levels of Porphyromonas gingivalis, Fusobacterium nucleatum and Prevotella intermedia in relation to oral leukoplakia (a potentially malignant disorder) and other participant characteristics. METHODS: This exploratory case-control study was nested within a large urban Indian cohort and the data included 22 men and women with oral leukoplakia (cases) and 69 leukoplakia-free controls. Each participant provided a salivary rinse sample, and a subset of 34 participants (9 cases; 25 controls) also provided a gingival swab sample from keratinized gingival surface for quantitative polymerase chain reaction (qPCR). RESULTS: Neither the distribution nor the levels of pathogens were associated with oral leukoplakia; however, individual pathogen levels were more strongly correlated with each other in cases compared to controls. Among controls, the median level of total pathogens was the highest (7.55×104 copies/ng DNA) among persons of low socioeconomic status. Salivary rinse provided better DNA concentration than gingival swab for qPCR analysis (mean concentration: 1.8 ng/µl vs. 0.2 ng/µl). CONCLUSIONS: This study confirms the feasibility of population studies evaluating oral microbiome in low-resource settings and identifies promising leads for future research.


Assuntos
DNA Bacteriano/genética , Fusobacterium nucleatum/isolamento & purificação , Leucoplasia Oral/microbiologia , Porphyromonas gingivalis/isolamento & purificação , Prevotella intermedia/isolamento & purificação , Adulto , Estudos de Casos e Controles , Países em Desenvolvimento , Estudos de Viabilidade , Feminino , Fusobacterium nucleatum/genética , Humanos , Índia , Masculino , Pessoa de Meia-Idade , Projetos Piloto , Porphyromonas gingivalis/genética , Prevotella intermedia/genética , Saliva/microbiologia , População Urbana
11.
PLoS One ; 16(3): e0242396, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33720954

RESUMO

The objective of this pilot study was to describe the microbial profiles present in the plaque and saliva of children who continued to develop new carious lesions following treatment with silver diamine fluoride ("nonresponders") compared to caries active, caries-free, and children immediately receiving SDF treatment for untreated caries in order to identify potential microbial differences that may relate to a re-incidence of caries. Saliva and plaque samples from infected and contralateral sites were obtained from twenty children who were either caries free, had active carious lesions, were caries active and received SDF treatment immediately before sampling, or had previously received SDF treatment and developed new caries. In total, 8,057,899 Illumina-generated sequence reads from 60 samples were obtained. Reads were processed using the Quantitative Insights Into Microbial Ecology pipeline. Group differences were assessed using Analysis of Variance Models and Tukey Honest Significant Differences. To identify significant taxa between treatment groups, Linear discriminant analysis Effect Size (LefSe) and Analysis of Differential Abundance Taking Sample Variation Into Account were used. Differential abundant analysis indicated that members of the Lachnospiraceae family were significantly enriched in non-responders and the genus Tannerella and species Granulicatella adiances were also highly abundant in this group. LefSe analysis between non-responders and SDF-treated groups revealed that genera Leptotrichia and Granulicatella were enriched in non-responders. We observed the highest abundance of phosphotransferase system and lowest abundance of lipopolysaccharide synthesis in non-responders. The microbiome in dental biofilms is responsible for initiation and progression of dental caries. SDF has been shown to be effective in arresting the progression carious lesions, in part due to its antimicrobial properties. Findings suggest that the differential abundance of select microbiota and specific pathway functioning in individuals that present with recurrent decay after SDF treatment may contribute to a potential failure of silver diamine fluoride to arrest dental caries. However, the short duration of sample collection following SDF application and the small sample size emphasize the need for further data and additional analysis.


Assuntos
Cárie Dentária/tratamento farmacológico , Microbiota , Compostos de Amônio Quaternário/uso terapêutico , Compostos de Prata/uso terapêutico , Carnobacteriaceae/genética , Carnobacteriaceae/isolamento & purificação , Criança , Estudos Transversais , DNA Bacteriano/química , DNA Bacteriano/metabolismo , Cárie Dentária/patologia , Placa Dentária/microbiologia , Análise Discriminante , Fluoretos Tópicos/uso terapêutico , Humanos , Leptotrichia/genética , Leptotrichia/isolamento & purificação , Projetos Piloto , Análise de Componente Principal , Saliva/microbiologia , Análise de Sequência de DNA , Falha de Tratamento
12.
Nat Microbiol ; 6(5): 574-583, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33737747

RESUMO

Mucus barriers accommodate trillions of microorganisms throughout the human body while preventing pathogenic colonization1. In the oral cavity, saliva containing the mucins MUC5B and MUC7 forms a pellicle that coats the soft tissue and teeth to prevent infection by oral pathogens, such as Streptococcus mutans2. Salivary mucin can interact directly with microorganisms through selective agglutinin activity and bacterial binding2-4, but the extent and basis of the protective functions of saliva are not well understood. Here, using an ex vivo saliva model, we identify that MUC5B is an inhibitor of microbial virulence. Specifically, we find that natively purified MUC5B downregulates the expression of quorum-sensing pathways activated by the competence stimulating peptide and the sigX-inducing peptide5. Furthermore, MUC5B prevents the acquisition of antimicrobial resistance through natural genetic transformation, a process that is activated through quorum sensing. Our data reveal that the effect of MUC5B is mediated by its associated O-linked glycans, which are potent suppressors of quorum sensing and genetic transformation, even when removed from the mucin backbone. Together, these results present mucin O-glycans as a host strategy for domesticating potentially pathogenic microorganisms without killing them.


Assuntos
Cárie Dentária/metabolismo , Mucina-5B/metabolismo , Polissacarídeos/metabolismo , Percepção de Quorum , Streptococcus mutans/fisiologia , Cárie Dentária/genética , Cárie Dentária/microbiologia , Interações Hospedeiro-Patógeno , Humanos , Mucina-5B/química , Mucina-5B/genética , Polissacarídeos/química , Saliva/metabolismo , Saliva/microbiologia , Streptococcus mutans/genética , Streptococcus mutans/patogenicidade , Transformação Bacteriana , Virulência
13.
NPJ Biofilms Microbiomes ; 7(1): 21, 2021 03 11.
Artigo em Inglês | MEDLINE | ID: mdl-33707430

RESUMO

Salivary gland epithelial cells (SGECs) have been implicated in the pathogenesis of Sjögren's syndrome due to aberrant antigen-presentation function. This study examined the hypothesis that oral dysbiosis modulates the antigen-presentation function of SGECs, which regulates CD4 T cell proliferation in primary Sjögren's syndrome (pSS). Saliva samples from 8 pSS patients and 16 healthy subjects were analyzed for bacterial 16S ribosomal DNA. As a result, 39 differentially abundant taxa were identified. Among them, the phylum Proteobacteria comprised 21 taxa, and this phylum was mostly enriched in the healthy controls. The proteobacterium Haemophilus parainfluenzae was enriched in the healthy controls, with the greatest effect size at the species level. Treatment of A253 cells in vitro with H. parainfluenzae upregulated PD-L1 expression, and H. parainfluenzae-pretreated A253 cells suppressed CD4 T cell proliferation. The suppression was partially reversed by PD-L1 blockade. Among low-grade xerostomia patients, salivary abundance of H. parainfluenzae decreased in pSS patients compared to that in non-pSS sicca patients. Our findings suggest that H. parainfluenzae may be an immunomodulatory commensal bacterium in pSS.


Assuntos
Disbiose/diagnóstico , Haemophilus parainfluenzae/imunologia , RNA Ribossômico 16S/genética , Saliva/microbiologia , Glândulas Salivares/citologia , Análise de Sequência de DNA/métodos , Síndrome de Sjogren/microbiologia , Idoso , Apresentação do Antígeno , Linfócitos T CD4-Positivos/metabolismo , Estudos de Casos e Controles , Linhagem Celular , Proliferação de Células , DNA Bacteriano/genética , DNA Ribossômico/genética , Células Epiteliais/citologia , Células Epiteliais/imunologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Filogenia , Glândulas Salivares/imunologia , Glândulas Salivares/microbiologia , Síndrome de Sjogren/imunologia
14.
J Microbiol Biotechnol ; 31(4): 501-509, 2021 Apr 28.
Artigo em Inglês | MEDLINE | ID: mdl-33746187

RESUMO

More than half the world's population is thought to be infected with Helicobacter pylori. Although the majority of infected people are asymptomatic, H. pylori infection may cause gastric ulcers and deadly gastric cancer. Owing to the difficulty and invasiveness of current routine culture and diagnostic methods, a highly sensitive and specific noninvasive assay for H. pylori is of interest. This study highlighted the design and performance of a colorimetric magneto loop-mediated isothermal amplification (CM-LAMP) assay to detect H. pylori in spiked saliva samples. LF primers were coated on magnetic nanoparticles by carbodiimide-induced immobilization and functionally used for solidphase amplification. During the LAMP reaction at 66°C, biotin-tagged FIPs were incorporated into LAMP amplicons. The colorimetric signal developed after the addition of NeutrAvidin horseradish peroxidase conjugate (NA-HRP) and ABTS. None of the tested microorganisms, including closely related bacteria, was shown positive by the CM-LAMP assay except H. pylori isolates. This novel platform was highly specific and 100-fold more sensitive (40 CFU/ml or 0.2 CFU per reaction) than the PCR and conventional LAMP assays for the detection of H. pylori in spiked saliva. Our results demonstrated the feasibility of using this noninvasive molecular diagnostic test to detect H. pylori in saliva samples.


Assuntos
Colorimetria , Infecções por Helicobacter/diagnóstico , Helicobacter pylori/isolamento & purificação , Técnicas de Diagnóstico Molecular , Técnicas de Amplificação de Ácido Nucleico , Humanos , Campos Magnéticos , Saliva/microbiologia , Sensibilidade e Especificidade
15.
J Photochem Photobiol B ; 217: 112168, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33714723

RESUMO

Worldwide shortages of personal protective equipment during COVID-19 pandemic has forced the implementation of methods for decontaminating face piece respirators such as N95 respirators. The use of UV irradiation to reduce bioburden of used respirators attracts attention, making proper testing protocols of uttermost importance. Currently artificial saliva is used but its comparison to human saliva from the UV disinfection perspective is lacking. Here we characterize UV spectra of human and artificial saliva, both fresh and after settling, to test for possible interference for UV-based disinfection. ASTM 2720 artificial saliva recipe (with either porcine or bovine mucin) showed many discrepancies from average (N = 18) human saliva, with different mucins demonstrating very different UV absorbance spectra, resulting in very different UV transmittance at different wavelength. Reducing porcine mucin concentration from 3 to 1.7 g/L brought UVA254 in the artificial saliva to that of average human saliva (although not for other wavelengths), allowing 254 nm disinfection experiments. Phosphate saline and modified artificial saliva were spiked with 8.6 log CFU/ml B. subtilis spores (ATCC 6633) and irradiated at dose of up to 100 mJ/cm2, resulting in 5.9 log inactivation for a saline suspension, and 2.8 and 1.1 log inactivation for ASTM-no mucin and ASTM-1.7 g/L porcine mucin 2 µL dried droplets, respectively. UVC irradiation of spores dried in human saliva resulted in 2.3 and 1.5 log inactivation, depending on the size of the droplets (2 vs 10 µL, respectively) dried on a glass surface. Our results suggest that in the presence of the current standard dried artificial saliva it is unlikely that UVC can achieve 6 log inactivation of B. subtilis spores using a realistic UV dose (e.g. less than 2 J/cm2) and the ATSM saliva recipe should be revised for UV decontamination studies.


Assuntos
Desinfecção/métodos , Saliva/química , Saliva/efeitos da radiação , Animais , Bacillus subtilis/efeitos da radiação , Canadá , Bovinos , Descontaminação/métodos , Feminino , Humanos , Israel , Masculino , Mucinas/química , Respiradores N95 , Saliva/microbiologia , Manejo de Espécimes/métodos , Espectrofotometria Ultravioleta , Esporos Bacterianos/efeitos da radiação , Raios Ultravioleta
16.
Medicine (Baltimore) ; 100(10): e24742, 2021 Mar 12.
Artigo em Inglês | MEDLINE | ID: mdl-33725829

RESUMO

ABSTRACT: Oral microbiota has been implicated in pathogenesis of recurrent aphthous stomatitis (RAS), which is a common mucosal disorder with unclear etiology. This study has explored the association between oral microbiota disorder and RAS in high-risk young female population.Forty-five young females were enrolled, including 24 RAS patients and 21 healthy individuals. Oral microbiome was analyzed by Illumina Miseq sequencing.Oral microbiota associated with RAS was characterized by the lower alpha-diversity indices (Chao1 and ACE). Several infectious pathogens increased in RAS, such as genera Actinobacillus, Haemophilus, Prevotella and Vibrio. The PICRUSt analysis indicated that the oral microbiota might be related with the up-regulation of genes involving infectious and neurodegenerative diseases, environmental adaptation, the down-regulation of genes involving basal metabolism, such as carbohydrate, energy, and amino acid metabolism.This study indicated that oral microbiota may play a significant role in RAS development.


Assuntos
Microbiota , Mucosa Bucal/microbiologia , Estomatite Aftosa/microbiologia , Feminino , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Recidiva , Saliva/microbiologia , Adulto Jovem
17.
Front Cell Infect Microbiol ; 11: 603291, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33718264

RESUMO

High serum thyroid-stimulating hormone (TSH) levels are linked to many metabolic disorders, but the effects of TSH levels on the oral microbiota are still largely unknown. This study aimed to explore the association between the salivary microbiome in adults and serum TSH levels. Saliva and fasting blood samples were obtained from a health census conducted in Southeast China. All participants were divided according to serum TSH levels. The microbial genetic profiles and changes were acquired by 16S rDNA sequencing and bioinformatics analysis. Relevant anthropometric and biochemical measurements such as insulin resistance, blood lipids, and body composition were evaluated with laboratory tests and physical examinations. The salivary microbiome in individuals with higher TSH level showed significantly higher taxa diversity. Principal coordinates analysis and partial least squares discriminant analysis showed distinct clustering in the Abnormal and Normal Groups (Adonis, P=0.0320). Granulicatella was identified as a discriminative genus for comparison of the two groups. Fasting serum insulin, Homeostatic Model Assessment for Insulin Resistance, and hemoglobin A1 were elevated in the Abnormal Group (P<0.05), showing the presence of insulin resistance in individuals with abnormal higher serum TSH levels. Distance-based redundancy analysis revealed the association of this distinctive difference with salivary microbiome. In conclusion, shifts in microbial profile were observed in the saliva of individuals with different serum TSH levels, and insulin resistance may play an important role in the biochemical and microbial alteration.


Assuntos
Resistência à Insulina , Microbiota , Saliva/microbiologia , Tireotropina/sangue , Adulto , China , Humanos
18.
Nutrients ; 13(3)2021 Feb 24.
Artigo em Inglês | MEDLINE | ID: mdl-33668205

RESUMO

The potent antimicrobial effects of antimicrobial photodynamic therapy (aPDT) with visible light plus water-filtered infrared-A irradiation and natural compounds as photosensitizers (PSs) have recently been demonstrated. The aim of this study was to obtain information on the antimicrobial effects of aPDT with mother juices against typical cariogenic oral Streptococcus pathogens in their planktonic form and determine its eradication potential on total human salivary bacteria from volunteers. Mother juices of pomegranate, bilberry, and chokeberry at different concentrations were used as PSs. The unweighted (absolute) irradiance was 200 mW cm-2, applied five minutes. Planktonic cultures of Streptococcus mutans and Streptococcus sobrinus and total mixed bacteria from pooled saliva of volunteers were treated with aPDT. Up to more than 5 log10 of S. mutans and S. sobrinus were killed by aPDT with 0.4% and 0.8% pomegranate juice, 3% and 50% chokeberry juice, and 12.5% bilberry juice (both strains). Concentrations of at least 25% (pomegranate) and >50% (chokeberry and bilberry) eradicated the mixed bacteria in saliva samples. This pilot study has shown that pomegranate mother juice is superior to the berry juices as a multicomponent PS for killing pathogenic oral bacteria with aPDT.


Assuntos
Antibacterianos/farmacologia , Sucos de Frutas e Vegetais/análise , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/farmacologia , Saliva/microbiologia , Humanos , Luz , Photinia/química , Projetos Piloto , Romã (Fruta)/química , Streptococcus mutans/efeitos dos fármacos , Streptococcus sobrinus/efeitos dos fármacos , Vaccinium myrtillus/química
19.
PLoS One ; 16(3): e0248308, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33667279

RESUMO

Periodontitis can result in tooth loss and the associated chronic inflammation can provoke several severe systemic health risks. Adjunctive to mechanical treatment of periodontitis and as alternatives to antibiotics, the use of probiotic bacteria was suggested. In this study, the inhibitory effect of the probiotic Streptococcus salivarius subsp. salivarius strains M18 and K12, Streptococcus oralis subsp. dentisani 7746, and Lactobacillus reuteri ATCC PTA 5289 on anaerobic periodontal bacteria and Aggregatibacter actinomycetemcomitans was tested. Rarely included in other studies, we also quantified the inverse effect of pathogens on probiotic growth. Probiotics and periodontal pathogens were co-incubated anaerobically in a mixture of autoclaved saliva and brain heart infusion broth. The resulting genome numbers of the pathogens and of the probiotics were measured by quantitative real-time PCR. Mixtures of the streptococcal probiotics were also used to determine their synergistic, additive, or antagonistic effects. The overall best inhibitor of the periodontal pathogens was L. reuteri ATCC PTA 5289, but the effect is coenzyme B12-, anaerobiosis-, as well as glycerol-dependent, and further modulated by L. reuteri strain DSM 17938. Notably, in absence of glycerol, the pathogen-inhibitory effect could even turn into a growth spurt. Among the streptococci tested, S. salivarius M18 had the most constant inhibitory potential against all pathogens, followed by K12 and S. dentisani 7746, with the latter still having significant inhibitory effects on P. intermedia and A. actinomycetemcomitans. Overall, mixtures of the streptococcal probiotics did inhibit the growth of the pathogens equally or-in the case of A. actinomycetemcomitans- better than the individual strains. P. gingivalis and F. nucleatum were best inhibited by pure cultures of S. salivarius K12 or S. salivarius M18, respectively. Testing inverse effects, the growth of S. salivarius M18 was enhanced when incubated with the periodontal pathogens minus/plus other probiotics. In contrast, S. oralis subsp. dentisani 7746 was not much influenced by the pathogens. Instead, it was significantly inhibited by the presence of other streptococcal probiotics. In conclusion, despite some natural limits such as persistence, the full potential for probiotic treatment is by far not utilized yet. Especially, further exploring concerted activity by combining synergistic strains, together with the application of oral prebiotics and essential supplements and conditions, is mandatory.


Assuntos
Anaerobiose/efeitos dos fármacos , Periodontite/tratamento farmacológico , Porphyromonas gingivalis/efeitos dos fármacos , Probióticos/farmacologia , Aggregatibacter actinomycetemcomitans/química , Aggregatibacter actinomycetemcomitans/crescimento & desenvolvimento , Antibiose/efeitos dos fármacos , Humanos , Lactobacillus reuteri/química , Lactobacillus reuteri/crescimento & desenvolvimento , Periodontite/microbiologia , Periodontite/patologia , Porphyromonas gingivalis/patogenicidade , Probióticos/química , Saliva/efeitos dos fármacos , Saliva/microbiologia , Streptococcus/química , Streptococcus/crescimento & desenvolvimento , Streptococcus mutans/química , Streptococcus mutans/crescimento & desenvolvimento , Streptococcus salivarius/química , Streptococcus salivarius/crescimento & desenvolvimento
20.
Br J Cancer ; 124(9): 1543-1551, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-33750907

RESUMO

BACKGROUND: Oral and gut microbiomes have emerged as potential biomarkers in cancer. We characterised the oral and gut microbiomes in a prospective observational cohort of HPV+ oropharyngeal squamous cell carcinoma (OPSCC) patients and evaluated the impact of chemoradiotherapy (CRT). METHODS: Saliva, oropharyngeal swabs over the tumour site and stool were collected at baseline and post-CRT. 16S RNA and shotgun metagenomic sequencing were used to generate taxonomic profiles, including relative abundance (RA), bacterial density, α-diversity and ß-diversity. RESULTS: A total of 132 samples from 22 patients were analysed. Baseline saliva and swabs had similar taxonomic composition (R2 = 0.006; p = 0.827). Oropharyngeal swabs and stool taxonomic composition varied significantly by stage, with increased oral RA of Fusobacterium nucleatum observed in stage III disease (p < 0.05). CRT significantly reduced the species richness and increased the RA of gut-associated taxa in oropharyngeal swabs (p < 0.05), while it had no effect in stool samples. These findings remained significant when adjusted by stage, smoking status and antibiotic use. CONCLUSIONS: Baseline oral and gut microbiomes differ by stage in this HPV+ cohort. CRT caused a shift towards a gut-like microbiome composition in oropharyngeal swabs. Stage-specific features and the transitions in oral microbiome might have prognostic and therapeutic implications.


Assuntos
Quimiorradioterapia/efeitos adversos , Microbioma Gastrointestinal , Mucosa Bucal/microbiologia , Neoplasias Orofaríngeas/terapia , Papillomaviridae/isolamento & purificação , Infecções por Papillomavirus/complicações , Saliva/microbiologia , Carcinoma de Células Escamosas/patologia , Carcinoma de Células Escamosas/terapia , Carcinoma de Células Escamosas/virologia , Feminino , Seguimentos , Humanos , Masculino , Mucosa Bucal/efeitos dos fármacos , Mucosa Bucal/efeitos da radiação , Neoplasias Orofaríngeas/patologia , Neoplasias Orofaríngeas/virologia , Infecções por Papillomavirus/virologia , Prognóstico , Estudos Prospectivos , Saliva/efeitos dos fármacos , Saliva/efeitos da radiação
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