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1.
Int J Mol Sci ; 21(14)2020 Jul 21.
Artigo em Inglês | MEDLINE | ID: mdl-32708341

RESUMO

Saliva is a highly versatile biological fluid that is easy to gather in a non-invasive manner-and the results of its analysis complement clinical and histopathological findings in the diagnosis of multiple diseases. The objective of this review was to offer an update on the contribution of salivary biomarkers to the diagnosis and prognosis of diseases of the oral cavity, including oral lichen planus, periodontitis, Sjögren's syndrome, oral leukoplakia, peri-implantitis, and medication-related osteonecrosis of the jaw. Salivary biomarkers such as interleukins, growth factors, enzymes, and other biomolecules have proven useful in the diagnosis and follow-up of these diseases, facilitating the early evaluation of malignization risk and the monitoring of disease progression and response to treatment. However, further studies are required to identify new biomarkers and verify their reported role in the diagnosis and/or prognosis of oral diseases.


Assuntos
Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Interleucinas/metabolismo , Boca/metabolismo , Saliva/metabolismo , Biomarcadores/metabolismo , Humanos , Leucoplasia Oral/diagnóstico , Leucoplasia Oral/enzimologia , Leucoplasia Oral/metabolismo , Líquen Plano Bucal/diagnóstico , Líquen Plano Bucal/enzimologia , Líquen Plano Bucal/metabolismo , Boca/enzimologia , Boca/patologia , Osteonecrose/diagnóstico , Osteonecrose/enzimologia , Osteonecrose/metabolismo , Peri-Implantite/diagnóstico , Peri-Implantite/enzimologia , Peri-Implantite/metabolismo , Periodontite/diagnóstico , Periodontite/enzimologia , Periodontite/metabolismo , Síndrome de Sjogren/diagnóstico , Síndrome de Sjogren/enzimologia , Síndrome de Sjogren/metabolismo
2.
Appl Environ Microbiol ; 81(16): 5471-6, 2015 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-26048943

RESUMO

Carbohydrate availability shifts when bacteria attach to a surface and form biofilm. When salivary planktonic bacteria form an oral biofilm, a variety of polysaccharides and glycoproteins are the primary carbon sources; however, simple sugar availabilities are limited due to low diffusion from saliva to biofilm. We hypothesized that bacterial glycoside hydrolase (GH) activities would be higher in a biofilm than in saliva in order to maintain metabolism in a low-sugar, high-glycoprotein environment. Salivary bacteria from 13 healthy individuals were used to grow in vitro biofilm using two separate media, one with sucrose and the other limiting carbon sources to a complex carbohydrate. All six GHs measured were higher in vitro when grown in the medium with complex carbohydrate as the sole carbon source. We then collected saliva and overnight dental plaque samples from the same individuals and measured ex vivo activities for the same six enzymes to determine how oral microbial utilization of glycoconjugates shifts between the planktonic phase in saliva and the biofilm phase in overnight dental plaque. Overall higher GH activities were observed in plaque samples, in agreement with in vitro observation. A similar pattern was observed in GH activity profiles between in vitro and ex vivo data. 16S rRNA gene analysis showed that plaque samples had a higher abundance of microorganisms with larger number of GH gene sequences. These results suggest differences in sugar catabolism between the oral bacteria located in the biofilm and those in saliva.


Assuntos
Glicosídeo Hidrolases/análise , Boca/microbiologia , Bactérias/classificação , Bactérias/genética , Biofilmes , Biota , DNA Ribossômico/química , DNA Ribossômico/genética , Voluntários Saudáveis , Humanos , Boca/enzimologia , RNA Ribossômico 16S/genética , Saliva/enzimologia , Saliva/microbiologia , Análise de Sequência de DNA
3.
Am J Pathol ; 182(2): 516-28, 2013 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-23219753

RESUMO

The functions of Rap-1A in oral carcinogenesis are largely unexplored. In this study, we examined the expression of Rap-1A at different malignant stages of oral cavity squamous cell carcinoma (OCSCC). Semiquantitative RT-PCR, quantitative RT-PCR, and Western blotting were used to evaluate Rap-1A mRNA and protein expressions, respectively, in paired OCSCC patient specimens. To determine the possible correlation between Rap-1A expression and various clinical characteristics, 256 samples from patients with OCSCC were evaluated by immunohistochemical staining. Strong Rap-1A expression was a significant prognostic marker and predictor of aggressive OCSCC. The overall and disease-specific 5-year survival rates were significantly correlated with strong expression of Rap-1A (P < 0.001). Functionally, overexpressed Rap-1A could promote oral cancer cell migration and invasion by Transwell chambers and wound healing assay. Conversely, the suppression of Rap-1A expression using Rap-1A-mediated siRNA was sufficient to decrease cell motility. Furthermore, our data also illustrated that Aurora-A could not only induce mRNA and protein expressions of Rap-1A for enhancing cancer cell motility but also co-localize and form a complex with Rap-1A in the oral cancer cell line. Finally, immunohistochemical staining, indirect immunofluorescence, and Western blotting analysis of human aggressive OCSCC specimens revealed a significantly positive correlation between Rap-1A and Aurora-A expression. Taken together, our results suggest that the Aurora-A/Rap-1A pathway is associated with survival, tumor progression, and metastasis of OCSCC patients.


Assuntos
Carcinoma de Células Escamosas/enzimologia , Carcinoma de Células Escamosas/patologia , Neoplasias Bucais/enzimologia , Neoplasias Bucais/patologia , Boca/patologia , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas rap1 de Ligação ao GTP/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Aurora Quinases , Carcinoma de Células Escamosas/genética , Linhagem Celular Tumoral , Movimento Celular , Feminino , Regulação Neoplásica da Expressão Gênica , Técnicas de Silenciamento de Genes , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Boca/enzimologia , Neoplasias Bucais/genética , Invasividade Neoplásica , Prognóstico , Modelos de Riscos Proporcionais , Ligação Proteica , RNA Interferente Pequeno/metabolismo , Análise de Regressão , Fatores de Risco , Análise de Sobrevida , Regulação para Cima/genética , Proteínas rap1 de Ligação ao GTP/genética
4.
ScientificWorldJournal ; 2014: 183548, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25389533

RESUMO

Betel quid (BQ) and areca nut (AN) (major BQ ingredient) are group I human carcinogens illustrated by International Agency for Research on Cancer and are closely associated with an elevated risk of oral potentially malignant disorders (OPMDs) and cancers of the oral cavity and pharynx. The primary alkaloid of AN, arecoline, can be metabolized via the monoamine oxidase (MAO) gene by inducing reactive oxygen species (ROS). The aim of this study was to investigate whether the variants of the susceptible candidate MAO genes are associated with OPMDs and oral and pharyngeal cancer. A significant trend of MAO-A mRNA expression was found in in vitro studies. Using paired human tissues, we confirmed the significantly decreased expression of MAO-A and MAO-B in cancerous tissues when compared with adjacent noncancerous tissues. Moreover, we determined that MAO-A single nucleotide polymorphism variants are significantly linked with oral and pharyngeal cancer patients in comparison to OPMDs patients [rs5953210 risk G-allele, odds ratio = 1.76; 95% confidence interval = 1.02-3.01]. In conclusion, we suggested that susceptible MAO family variants associated with oral and pharyngeal cancer may be implicated in the modulation of MAO gene activity associated with ROS.


Assuntos
Arecolina/toxicidade , Carcinoma de Células Escamosas/genética , Monoaminoxidase/genética , Neoplasias Bucais/genética , Neoplasias Faríngeas/genética , RNA Mensageiro/genética , Areca/química , Arecolina/metabolismo , Carcinoma de Células Escamosas/enzimologia , Carcinoma de Células Escamosas/patologia , Expressão Gênica , Humanos , Monoaminoxidase/metabolismo , Boca/enzimologia , Boca/patologia , Neoplasias Bucais/enzimologia , Neoplasias Bucais/patologia , Neoplasias Faríngeas/enzimologia , Neoplasias Faríngeas/patologia , Faringe/enzimologia , Faringe/patologia , Polimorfismo de Nucleotídeo Único , RNA Mensageiro/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Risco , Microambiente Tumoral
5.
Eksp Klin Gastroenterol ; (3): 61-6, 2014.
Artigo em Russo | MEDLINE | ID: mdl-25518484

RESUMO

STUDY OBJECTIVE: Analysis of the prevalence of mouth cavity urealytic microflora and determination of the level of its enzymatic activity depending on concentration and amount of urea solution taken as a substrate. MATERIALS AND METHODS: 62 randomly chosen patients at the age of 5-64 took part in the study. Each of them rinsed the mouth with 50 ml of 1% urea solution. Before and after rinsing the concentration of ammonia in the mouth cavity air was measured. In patients with highest and lowest activity of mouth cavity urealytic microflora a series of tests was carried out including mouth rinsing with urea solution in various concentrations and amounts and measuring ammonia concentration before and after rinsing. Obtained results were analyzed using mathematical statistics methods. RESULTS: It was found that in 91% ± 1.8% of randomly chosen patients (p < 0.05) mouth cavity microflora showed apparent urease activity. The lowest concentration (0.0625% in 50 ml) and volume (0.5 ml of 1% solution) levels of urea solution were obtained that can exert negative influence on the results of helicobacteriosis diagnosis by means of mouth cavity air analysis. CONCLUSION: Urealytic microflora in the mouth cavity is very common and may constitute a factor that decreases the specificity of helicobacteriosis diagnosis by means of the methods based on detection of indicators of gas metabolites resulting from the enzymatic reaction in air samples taken from the mouth cavity after oral administration of urea.


Assuntos
Proteínas de Bactérias/metabolismo , Infecções por Helicobacter/diagnóstico , Infecções por Helicobacter/enzimologia , Helicobacter pylori/enzimologia , Boca , Urease/metabolismo , Adolescente , Adulto , Criança , Pré-Escolar , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Boca/enzimologia , Boca/microbiologia
6.
ScientificWorldJournal ; 2013: 920595, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23365550

RESUMO

One hallmark of cancer is the degradation of the extracellular matrix (ECM), which is caused by proteinases. In oral cancers, matrix metalloproteinases (MMPs), especially MMP-9, are associated with this degradation. MMPs break down the ECM allowing cancer to spread; they also release various factors from their cryptic sites, including cytokines. These factors modulate cell behavior and enhance cancer progression by regulating angiogenesis, migration, proliferation, and invasion. The development of early metastases is typical for oral cancer, and increased MMP-9 expression is associated with a poor disease prognosis. However, many studies fail to relate MMP-9 expression with metastasis formation. Contrary to earlier models, recent studies show that MMP-9 plays a protective role in oral cancers. Therefore, the role of MMP-9 is complicated and may fluctuate throughout the different types and stages of oral cancers.


Assuntos
Carcinoma de Células Escamosas/enzimologia , Metaloproteinase 9 da Matriz/metabolismo , Modelos Biológicos , Neoplasias Bucais/enzimologia , Boca/enzimologia , Animais , Humanos
7.
Plant Physiol ; 156(3): 1520-34, 2011 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-21546453

RESUMO

How plants perceive herbivory is not yet well understood. We investigated early responses of the model plant Arabidopsis (Arabidopsis thaliana) to attack from the generalist grasshopper herbivore, Schistocerca gregaria (Caelifera). When compared with wounding alone, S. gregaria attack and the application of grasshopper oral secretions (GS) to puncture wounds elicited a rapid accumulation of various oxylipins, including 13-hydroperoxy octadecatrienoic acid, 12-oxo-phytodienoic acid (OPDA), jasmonic acid, and jasmonic acid-isoleucine. Additionally, GS increased cytosolic calcium levels, mitogen-activated protein kinase (MPK3 and MPK6) activity, and ethylene emission but not the accumulation of hydrogen peroxide. Although GS contain caeliferin A16:0, a putative elicitor of caeliferan herbivores, treatment with pure, synthetic caeliferin A16:0 did not induce any of the observed responses. With mutant plants, we demonstrate that the observed changes in oxylipin levels are independent of MPK3 and MPK6 activity but that MPK6 is important for the GS-induced ethylene release. Biochemical and pharmacological analyses revealed that the lipase activity of GS plays a central role in the GS-induced accumulation of oxylipins, especially OPDA, which could be fully mimicked by treating puncture wounds only with a lipase from Rhizopus arrhizus. GS elicitation increased the levels of OPDA-responsive transcripts. Because the oral secretions of most insects used to study herbivory-induced responses in Arabidopsis rapidly elicit similar accumulations of OPDA, we suggest that lipids containing OPDA (arabidopsides) play an important role in the activation of herbivory-induced responses.


Assuntos
Arabidopsis/imunologia , Insetos/enzimologia , Lipase/metabolismo , Boca/enzimologia , Animais , Arabidopsis/efeitos dos fármacos , Arabidopsis/enzimologia , Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Ciclopentanos/metabolismo , Etilenos/metabolismo , Ácidos Graxos Insaturados/farmacologia , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Gafanhotos/efeitos dos fármacos , Gafanhotos/enzimologia , Insetos/efeitos dos fármacos , Quinases de Proteína Quinase Ativadas por Mitógeno/metabolismo , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Dados de Sequência Molecular , Oxilipinas/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Espécies Reativas de Oxigênio/metabolismo
8.
Dig Dis Sci ; 57(2): 413-8, 2012 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-21881975

RESUMO

BACKGROUND: Despite the rich literature on GERD, its cause and reason for increased prevalence remain obscure. Currently accepted mechanisms leave many questions unanswered. Nitrite chemistry at the GEJ is well described for carcinogenesis. Recent epidemiological and animal data have linked nitrates to GERD. "Nitrate reductase" of oral bacteria converts nitrates to nitrites. We hypothesized that nitrate reductase activity is higher in patients with erosive GERD, delivering more nitrite at the gastroesophageal-junction for a given nitrate intake. AIM: To compare oral nitrate reductase activity of erosive GERD patients with controls. METHODS: Patients with erosive GERD and controls without GERD were enrolled. After overnight fasting, nitrite of oral cavity contents was measured at 1-min intervals for 3 min while incubating a 10-mg nitrate-N/L solution in the mouth. Nitrate reductase activity was calculated and compared between groups. RESULTS: Eleven cases (ten males, mean age: 42.6 ± 11.7 year) and ten controls (eight males, mean age: 37.6 ± 9.2 year) were enrolled. Mean nitrate reductase activity was 3.23 ± 0.99 vs. 2.30 ± 0.83 "µg nitrite-N formed/person/minute" in cases and controls, respectively (p = 0.03). CONCLUSIONS: Oral nitrate reductase activity in erosive GERD patients is higher than controls. Therefore, any dietary nitrate load generates more nitrite in these patients. This excess nitrite at the gastroesophageal junction, may potentially contribute to the development of GERD. This is the first report linking oral nitrite production to erosive GERD in man. We suggest that a "nitrate hypothesis" may answer yet unanswered questions about GERD pathogenesis. If confirmed, it may change our understanding of mechanisms of GERD and provide novel therapeutic targets.


Assuntos
Refluxo Gastroesofágico/fisiopatologia , Boca/enzimologia , Nitrato Redutase/metabolismo , Adulto , Feminino , Refluxo Gastroesofágico/metabolismo , Humanos , Masculino , Pessoa de Meia-Idade , Nitritos
9.
Adv Dent Res ; 24(2): 77-80, 2012 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-22899685

RESUMO

Recent rapid advances in "-omics" technologies have yielded new insights into the interaction of the oral microbiome with its host. Associations of species that are usually considered to be acid-tolerant with caries have been confirmed, while some recognized as health-associated are often present in greater proportions in the absence of caries. In addition, some newly identified bacteria have been suggested as potential contributors to the caries process. In spite of this progress, two major challenges remain. The first is that there is a great deal of heterogeneity in the phenotypic capabilities of individual species of oral bacteria. The second is that the most abundant taxa in oral biofilms display remarkable phenotypic plasticity, i.e., the bacteria associated most strongly with health or with caries can morph rapidly in response to alterations in environmental pH, carbohydrate availability and source, and oxygen tension and redox environment. However, new technologic advances coupled with "old-fashioned microbiology" are starting to erode the barriers to a more complete understanding of oral biofilm physiology and ecology, and in doing so are beginning to provide insights for the creation of novel cost-effective caries control therapies.


Assuntos
Cárie Dentária/microbiologia , Metagenoma/genética , Boca/microbiologia , Streptococcus mutans/patogenicidade , Arginina/genética , Arginina/metabolismo , Biofilmes/classificação , Cárie Dentária/prevenção & controle , Cárie Dentária/terapia , Humanos , Metagenoma/fisiologia , Boca/enzimologia , RNA Ribossômico 16S/análise , Streptococcus mutans/genética
10.
Oxid Med Cell Longev ; 2022: 1886277, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35116090

RESUMO

Oral diseases are among the most common human diseases yet less studied. These diseases affect both the physical, mental, and social health of the patients resulting in poor quality of life. They affect all ages, although severe stages are mostly observed in older individuals. Poor oral hygiene, genetics, and environmental factors contribute enormously to the development and progression of these diseases. Although there are available treatment options for these diseases, the recurrence of the diseases hinders their efficiency. Oral volatile sulfur compounds (VSCs) are highly produced in oral cavity as a result of bacteria activities. Together with bacteria components such as lipopolysaccharides, VSCs participate in the progression of oral diseases by regulating cellular activities and interfering with the immune response. Hydrogen sulfide (H2S) is a gaseous neurotransmitter primarily produced endogenously and is involved in the regulation of cellular activities. The gas is also among the VSCs produced by oral bacteria. In numerous diseases, H2S have been reported to have dual effects depending on the cell, concentration, and donor used. In oral diseases, high production and subsequent utilization of this gas have been reported. Also, this high production is associated with the progression of oral diseases. In this review, we will discuss the production of H2S in oral cavity, its interaction with cellular activities, and most importantly its role in oral diseases.


Assuntos
Sulfeto de Hidrogênio/metabolismo , Doenças da Boca/patologia , Apoptose , Bactérias/isolamento & purificação , Bactérias/metabolismo , Cistationina gama-Liase/metabolismo , Humanos , Boca/enzimologia , Boca/metabolismo , Boca/microbiologia , Doenças da Boca/metabolismo , Neoplasias Bucais/metabolismo , Neoplasias Bucais/patologia , Estresse Oxidativo
11.
Microb Pathog ; 50(3-4): 148-54, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21238567

RESUMO

Streptococcus sanguinis is a member of oral streptococci and one of the most abundant species found in oral biofilm called dental plaque. Colonization of the oral streptococci on the tooth surface depends on the adhesion of bacteria to salivary components adsorbed to the tooth surface. Recently, we identified unique cell surface long filamentous structures named pili in this species. Herein, we investigated the role of S. sanguinis pili in biofilm formation. We found that pili-deficient mutant, in which the genes encoding the three pilus proteins PilA, PilB and PilC have been deleted, showed an impaired bacterial accumulation on saliva-coated surfaces. Confocal microscopic observations suggested that the mutant was incapable of producing typical three-dimensional layer of biofilm. Ligand blot analysis showed that the ancillary pilus proteins PilB and PilC bound to human whole saliva. Additional analysis demonstrated that PilC bound to multiple salivary components, and one of which was found to be salivary α-amylase. These results indicate that pilus proteins are members of saliva-binding proteins of oral S. sanguinis, and suggest the involvement of pili in its colonization on saliva-coated tooth surfaces and in the human oral cavity.


Assuntos
Amilases/metabolismo , Biofilmes , Fímbrias Bacterianas/metabolismo , Saliva/enzimologia , Infecções Estreptocócicas/enzimologia , Streptococcus sanguis/fisiologia , Amilases/genética , Aderência Bacteriana , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Proteínas de Fímbrias/genética , Proteínas de Fímbrias/metabolismo , Fímbrias Bacterianas/genética , Humanos , Boca/enzimologia , Boca/microbiologia , Ligação Proteica , Saliva/microbiologia , Infecções Estreptocócicas/microbiologia , Streptococcus sanguis/genética
12.
Arch Microbiol ; 193(12): 905-10, 2011 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-21892611

RESUMO

To better understand the phenomena governing the establishment of the oral bacterium Streptococcus salivarius in the mouth, the effect of some environmental factors has been studied in complemented artificial saliva, under oral pH and temperature conditions. Three salivary enzymes at physiological concentrations were tested: peroxidase, lysozyme and amylase, as well as injection of exhaled air. Injection of air containing 5% CO2 and 16% O2 induced a deleterious effect on S. salivarius K12, mainly by increasing redox potential. Addition of lysozyme slightly affected the physiological state of S. salivarius by altering membrane integrity. In contrast, peroxidase was not detrimental as it made it possible to decrease the redox potential. The addition of amylase reduced the specific growth rate of S. salivarius by formation of a complex with amylase and mucins, but led to high final biomass, as a result of enzymatic degradation of some nutrients. Finally, this work demonstrated that salivary enzymes had a slight impact on S. salivarius behaviour. It can thus be concluded that this bacterium was well adapted to in-mouth conditions, as it was able to resist certain salivary enzymes, even if tolerance to expired air was affected, as a result of an increased redox potential.


Assuntos
Ar/análise , Saliva Artificial/metabolismo , Saliva/enzimologia , Streptococcus/crescimento & desenvolvimento , Amilases/metabolismo , Membrana Celular/fisiologia , Viabilidade Microbiana , Boca/enzimologia , Boca/microbiologia , Muramidase/metabolismo , Oxirredução , Peroxidase/metabolismo , Saliva/microbiologia , Streptococcus/fisiologia
13.
Mol Med Rep ; 24(1)2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-33982776

RESUMO

Sialoperoxidase and myeloperoxidase are the two main peroxidase enzymes found in the oral cavity. Sialoperoxidase is present in salivary secretions and in the biofilms that line the oral surfaces, while myeloperoxidase is abundant in the dento­gingival sulcus area. In the presence of hydrogen peroxide (H2O2), oral peroxidases catalyze the oxidation of the pseudohalide anion thiocyanate (SCN­) to hypothiocyanite (OSCN­), a strong oxidant that serves an antimicrobial role. Furthermore, oral peroxidases consume bacteria­produced H2O2 and could help inactivate toxic carcinogenic and genotoxic substances. Numerous in vitro studies have reported the antibacterial, antimycotic and antiviral role of peroxidases, suggesting possible applications in oral therapy. However, the use of oral hygiene products incorporating peroxidase systems has not yet been shown to be beneficial for the treatment or prevention of oral infections. This paradox reflects our incomplete knowledge of the physiological role of peroxidases in a complex environment, such as the oral region. While hygiene is crucial for restoring oral microbiota to a symbiotic state, there are no data to suggest that the addition of a peroxidase per se can create a dysbiotic state. Recent investigations have associated the presence of peroxidase activity with gram­positive cocci microbial flora, and its insufficiency with dysbiosis has been linked to pathologies, such as caries, periodontitis or infections of the oral mucosa. Therefore, oxidants generated by oral peroxidases appear to be an essential ecological determinant for oral health through the selection of a symbiotic microbiota capable of resisting oxidative stress. The objective of the present review was to update the current knowledge of the physiological aspects and applications of oral peroxidases in clinical practice.


Assuntos
Anti-Infecciosos/farmacologia , Anti-Infecciosos/uso terapêutico , Boca/enzimologia , Boca/microbiologia , Higiene Bucal/métodos , Peroxidases/farmacologia , Peroxidases/uso terapêutico , Animais , Mimetismo Biológico , Humanos , Oxidantes/metabolismo
14.
J Electron Microsc (Tokyo) ; 59(4): 311-20, 2010.
Artigo em Inglês | MEDLINE | ID: mdl-20388619

RESUMO

Bacterial virulence could be altered by the antimicrobial agents of the host. Our aim was to identify the damage and survival of Streptococcus sanguinis induced by lysozymes in vitro and to analyse the potential of oral microorganisms to shirk host defences, which cause infective endocarditis. S. sanguinis ATCC 10556 received lysozyme at concentrations of 12.5, 25, 50 and 100 microg/ml. Cells were examined by electron microscopy. The survival was assessed by colony counting and construction of a growth curve. Challenged by lysozymes, cells mainly exhibited cell wall damage, which seemed to increase with increasing lysozyme concentration and longer incubation period in the presence of ions. Cells with little as well as apparent lesion were observed under the same treatment set, and anomalous stick and huge rotund bodies were occasionally observed. After the removal of the lysozyme, some damaged cells could be reverted to its original form with brain heart infusion (BHI), and their growth curve was similar to the control cells. After further incubation in BHI containing lysozyme, S. sanguinis cell damage stopped progressing, and their growth curve was also similar to the control cells. The results suggested that the S. sanguinis lesions caused by the lysozyme in the oral cavity may be nonhomogeneous and that some damaged cells could self-repair and survive. It also indicated that S. sanguinis with damaged cell walls may survive and be transmitted in the bloodstream.


Assuntos
Anti-Infecciosos/farmacologia , Muramidase/farmacologia , Streptococcus sanguis/efeitos dos fármacos , Streptococcus sanguis/ultraestrutura , Animais , Endocardite Bacteriana/microbiologia , Humanos , Viabilidade Microbiana , Microscopia Eletrônica/métodos , Boca/enzimologia , Boca/microbiologia , Infecções Estreptocócicas/microbiologia , Streptococcus sanguis/crescimento & desenvolvimento
15.
J Tradit Chin Med ; 30(2): 126-31, 2010 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-20653170

RESUMO

OBJECTIVE: To explore the influence of charred Gossamer urocteae (CGU) on the functions of primary cultured mouse oral fibroblasts and reveal its mechanism in wound healing. METHODS: CGU was extracted with different solvents and ethanol extract (EE), ethyl acetate fraction (EF), n-butanol fraction (BF) and aqueous fraction (AF) were obtained. The effects of different fractions on the proliferation, matrix metalloproteinase-2,9 (MMP-2,9) activities, synthesis of collagen and tissue inhibitor of metalloproteinase 1 (TIMP-1) in the mouse oral fibroblasts were determined by MTT, gelatin zymography, chloramine-T method, and enzyme-linked immunosorbent assay (ELISA) respectively. RESULTS: EE, EF and BF at high concentrations could significantly inhibit proliferation of fibroblasts (P<0.05 or P<0.01), and at low concentrations EF and BF could promote proliferation of fibroblasts, and BF and AF could significantly inhibit collagen synthesis (P<0.05 or P<0.01). EE, EF and AF at high concentrations could significantly increase the MMP-9 activity, and BF and AF could significantly inhibit synthesis of TIMP-1. CONCLUSION: CGU at high concentrations can inhibit the proliferations of fibroblasts and synthesis of collagen, and in healing of wound, CGU at high concentrations possibly has the functions of anti-fibrosis and anti-scar, and the mechanism to promote degradation of collagen is possibly related to the increase in MMP-9 activity and the inhibition of TIMP-1 synthesis.


Assuntos
Fatores Biológicos/farmacologia , Fibroblastos/efeitos dos fármacos , Medicina Tradicional Chinesa , Boca/citologia , Aranhas/química , Animais , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Colágeno/metabolismo , Fibroblastos/enzimologia , Fibroblastos/metabolismo , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Boca/efeitos dos fármacos , Boca/enzimologia , Boca/metabolismo , Inibidor Tecidual de Metaloproteinase-1/metabolismo
16.
Zhonghua Kou Qiang Yi Xue Za Zhi ; 55(5): 353-356, 2020 May 09.
Artigo em Zh | MEDLINE | ID: mdl-32392980

RESUMO

Nitric oxide (NO) is a short-lived free radical which is not only involved in regulating many physiological processes of the body, but also closely related to many diseases. Nitric oxide synthase (NOS) is the key enzyme for NO production. NOS exists as three distinct isoforms, the endothelial NOS (eNOS), neuronal NOS (nNOS) and inducible NOS (iNOS). It has been found that nNOS and eNOS were expressed in normal pulp tissues, periodontal tissues and salivary glands, and the NO produced from nNOS and eNOS was involved in their physiological functions. NO and iNOS are involved in the occurrence and development of pulpitis, periodontitis, salivary gland disease and oral cancer. This review focuses on the physiological and pathological effects of NO and different subtypes of NOS in oral cavity.


Assuntos
Doenças da Boca/enzimologia , Boca/enzimologia , Boca/fisiopatologia , Óxido Nítrico Sintase/fisiologia , Óxido Nítrico/fisiologia , Humanos , Doenças da Boca/fisiopatologia
17.
J Oral Pathol Med ; 38(7): 591-6, 2009 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-19453842

RESUMO

BACKGROUND: The human oral cells have different physiological properties and different origins in developmental stages. Mechanical, physiological, and chemical stress can cause damage and irritation during clinical treatment in various oral tissues. PURPOSE: The effects of DNA damage response and gene silencing of checkpoint kinases (Chk1/2) is not unclear in oral primary and cancer cells. METHOD: Treatment with doxorubicin involving DNA damage and gene silencing of Chk1/2 by shRNA constructs was performed in pulp, periodontal ligament, gingival tissues (HGF), and mouth epithelial carcinoma cells (KB). RESULTS: The KB cells were more sensitive to genotoxic stress response than oral primary cells. Endogenous levels of Chk1/2 in KB cell were higher than in pulp cells. When doxorubicin was administered, Chk2 activation was induced in KB cell, but not in pulp cells. However, viability in KB cells did not decrease by the suppression of the checkpoint proteins, whereas primary cells were defective in gene silencing. When doxorubicin treatment and gene silencing were combined, both primary cells and KB cell were defective. Moreover, in case of KB cell, cell death was increased and activation of Chk2 was increased in doxorubicin dose-dependent. CONCLUSION: These data indicate that not only stress response mechanism may be different in oral primary and cancer cells but also Chk1/2 proteins may have essential roles in oral primary cells. Based on these data, checkpoint proteins may be crucial drug targets for oral cancer therapy.


Assuntos
Carcinoma/enzimologia , Citotoxinas/toxicidade , Doxorrubicina/toxicidade , Fibroblastos/enzimologia , Neoplasias Bucais/enzimologia , Proteínas Serina-Treonina Quinases/metabolismo , Carcinoma/tratamento farmacológico , Carcinoma/genética , Carcinoma/patologia , Sobrevivência Celular , Quinase 1 do Ponto de Checagem , Quinase do Ponto de Checagem 2 , Dano ao DNA/fisiologia , Polpa Dentária/citologia , Polpa Dentária/efeitos dos fármacos , Polpa Dentária/enzimologia , Relação Dose-Resposta a Droga , Fibroblastos/efeitos dos fármacos , Inativação Gênica , Genes cdc/efeitos dos fármacos , Genes cdc/fisiologia , Gengiva/efeitos dos fármacos , Gengiva/enzimologia , Humanos , Boca/citologia , Boca/efeitos dos fármacos , Boca/enzimologia , Neoplasias Bucais/tratamento farmacológico , Neoplasias Bucais/genética , Neoplasias Bucais/patologia , Ligamento Periodontal/efeitos dos fármacos , Ligamento Periodontal/enzimologia , Proteínas Quinases/efeitos dos fármacos , Proteínas Quinases/genética , Proteínas Quinases/metabolismo , Proteínas Serina-Treonina Quinases/efeitos dos fármacos , Proteínas Serina-Treonina Quinases/genética , Células Tumorais Cultivadas
18.
Nat Protoc ; 14(4): 991-1014, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30886367

RESUMO

Developing a mechanistic understanding of the impact of food structure and composition on human health has increasingly involved simulating digestion in the upper gastrointestinal tract. These simulations have used a wide range of different conditions that often have very little physiological relevance, and this impedes the meaningful comparison of results. The standardized protocol presented here is based on an international consensus developed by the COST INFOGEST network. The method is designed to be used with standard laboratory equipment and requires limited experience to encourage a wide range of researchers to adopt it. It is a static digestion method that uses constant ratios of meal to digestive fluids and a constant pH for each step of digestion. This makes the method simple to use but not suitable for simulating digestion kinetics. Using this method, food samples are subjected to sequential oral, gastric and intestinal digestion while parameters such as electrolytes, enzymes, bile, dilution, pH and time of digestion are based on available physiological data. This amended and improved digestion method (INFOGEST 2.0) avoids challenges associated with the original method, such as the inclusion of the oral phase and the use of gastric lipase. The method can be used to assess the endpoints resulting from digestion of foods by analyzing the digestion products (e.g., peptides/amino acids, fatty acids, simple sugars) and evaluating the release of micronutrients from the food matrix. The whole protocol can be completed in ~7 d, including ~5 d required for the determination of enzyme activities.


Assuntos
Materiais Biomiméticos/metabolismo , Ingredientes de Alimentos/análise , Intestinos/enzimologia , Modelos Biológicos , Boca/enzimologia , Estômago/enzimologia , Aminoácidos/análise , Aminoácidos/química , Bile/enzimologia , Materiais Biomiméticos/química , Digestão/fisiologia , Ingestão de Alimentos/fisiologia , Ensaios Enzimáticos/normas , Ácidos Graxos/análise , Ácidos Graxos/química , Alimentos , Suco Gástrico/enzimologia , Humanos , Concentração de Íons de Hidrogênio , Hidrólise , Oligossacarídeos/análise , Oligossacarídeos/química , Fragmentos de Peptídeos/análise , Fragmentos de Peptídeos/química , Saliva/enzimologia
19.
Gene Expr Patterns ; 8(4): 284-90, 2008 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-18203667

RESUMO

Investigations into molecular mechanisms in vertebrates have examined which growth factors regulate many of the essential underlying cellular processes in development. Growth factors regulate cell proliferation and differentiation through diverse signaling pathways like the MEK (mitogen-activated protein kinase) and ERK (extracellular signal-regulated kinase) pathway. The MEK and ERK pathway can interact with the PI3K (phosphatidylinositol-3-kinase) and PTEN (phosphatase and tensin homologues deleted on chromosome 10) signaling pathway. Interactions between these pathways during development have been extensively studied in many organs; however, the importance of these pathways in oral development is not well known. In this study, we examined the expression of the phosphorylated forms of ERK (pERK), MEK (pMEK), PTEN (pPTEN) and PI3K during mouse development from E13.5 to E16.5. We found unique and overlapping expression of these factors in the craniofacial region, with pERK and pPTEN showing opposing activation patterns in both the tooth and the tongue.


Assuntos
MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Quinases de Proteína Quinase Ativadas por Mitógeno/metabolismo , Boca/embriologia , PTEN Fosfo-Hidrolase/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Animais , Imuno-Histoquímica , Camundongos , Boca/enzimologia , Palato/embriologia , Palato/enzimologia , Fosforilação , Língua/embriologia , Língua/enzimologia , Dente/embriologia , Dente/enzimologia
20.
Contrib Microbiol ; 15: 164-187, 2008.
Artigo em Inglês | MEDLINE | ID: mdl-18511861

RESUMO

The importance of reactive oxygen species (ROS) in innate immunity was first recognized in professional phagocytes undergoing a 'respiratory burst'upon activation. This robust oxygen consumption is related to a superoxide-generating enzyme, the phagocytic NADPH oxidase (Nox2-based or phox). The oxidase is essential for microbial killing, since patients lacking a functional oxidase suffer from enhanced susceptibility to microbial infections. ROS derived from superoxide attack bacteria in the isolated niche of the neutrophil phagosome. The oxidase is electrogenic, alters ion currents across membranes, induces apoptosis, regulates cytokine production, influences gene expression, and promotes formation of extracellular traps. Recently, new homologues of Nox2 were discovered establishing the Nox family of NADPH oxidases that encompasses seven members. Nox1 is highly expressed in the colon epithelium, and can be induced by LPS or IFN- gamma. Nox4 was implicated in innate immunity since LPS induces Nox4-dependent ROS generation. Duox1 and Duox2 localize to the apical plasma membrane of epithelial cells in major airways, salivary glands, and the gastrointestinal tract, and provide extracellular hydrogen peroxide to lactoperoxidase to produce antimicrobial hypothiocyanite ions. Th1 and Th2 cytokines regulate expression of dual oxidases in human airways and may thereby act in host defense or in proinflammatory responses.


Assuntos
Imunidade Inata , NADPH Oxidases/imunologia , Espécies Reativas de Oxigênio/imunologia , Animais , Trato Gastrointestinal/enzimologia , Trato Gastrointestinal/imunologia , Trato Gastrointestinal/microbiologia , Expressão Gênica , Humanos , Boca/enzimologia , Boca/imunologia , Boca/microbiologia , Família Multigênica , NADPH Oxidases/química , NADPH Oxidases/genética , NADPH Oxidases/metabolismo , Neutrófilos/enzimologia , Neutrófilos/imunologia , Neutrófilos/microbiologia , Oxigênio/metabolismo , Fagocitose , Fagossomos/enzimologia , Sistema Respiratório/enzimologia , Sistema Respiratório/imunologia , Sistema Respiratório/microbiologia
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