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1.
Mol Oral Microbiol ; 33(1): 47-58, 2018 02.
Artigo em Inglês | MEDLINE | ID: mdl-28845576

RESUMO

The oral bacterium Streptococcus mutans is the principal agent in the development of dental caries. Biofilm formation by S. mutans requires bacterial attachment, aggregation, and glucan formation on the tooth surface under sucrose supplementation conditions. Our previous microarray analysis of clinical strains identified 74 genes in S. mutans that were related to biofilm morphology; however, the roles of almost all of these genes in biofilm formation are poorly understood. We investigated the effects of 21 genes randomly selected from our previous study regarding S. mutans biofilm formation, regulation by the complement pathway, and responses to competence-stimulating peptide. Eight competence-stimulating peptide-dependent genes were identified, and their roles in biofilm formation and aggregation were examined by mutational analyses of the S. mutansUA159 strain. Of these eight genes, the inactivation of the putative hemolysin III family SMU.940 gene of S. mutansUA159 promoted rapid dextran-dependent aggregation and biofilm formation in tryptic soy broth without dextrose (TSB) with 0.25% glucose and slightly reduced biofilm formation in TSB with 0.25% sucrose. The SMU.940 mutant showed higher expression of GbpC and gbpC gene than wild-type. GbpC is known to be involved in the dextran-dependent aggregation of S. mutans. An SMU.940-gbpC double mutant strain was constructed in the SMU.940 mutant background. The gbpC mutation completely abolished the dextran-dependent aggregation of the SMU.940 mutant. In addition, the aggregation of the mutant was abrogated by dextranase. These findings suggest that SMU.940 controls GbpC expression, and contributes to the regulation of dextran-dependent aggregation and biofilm formation.


Assuntos
Adesinas Bacterianas/genética , Proteínas de Bactérias/genética , Biofilmes/crescimento & desenvolvimento , Dextranos/metabolismo , Genes Bacterianos/genética , Streptococcus mutans/genética , Streptococcus mutans/metabolismo , Adesinas Bacterianas/metabolismo , Aderência Bacteriana/genética , Proteínas de Transporte/genética , Proteínas de Transporte/metabolismo , Análise Mutacional de DNA , Perfilação da Expressão Gênica , Regulação Bacteriana da Expressão Gênica , Mutação , Polissacarídeos/análise , RNA Bacteriano/análise , Streptococcus mutans/crescimento & desenvolvimento
2.
Mol Med Rep ; 17(1): 843-851, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-29115611

RESUMO

Dental caries is an important global health concern and Streptococcus mutans has been established as a major cariogenic bacterial species. Reports indicate that a rare sugar, D­tagatose, is not easily catabolized by pathogenic bacteria. In the present study, the inhibitory effects of D­tagatose on the growth and biofilm formation of S. mutans GS­5 were examined. Monitoring S. mutans growth over a 24 h period revealed that D­tagatose prolonged the lag phase without interfering with the final cell yield. This growth retardation was also observed in the presence of 1% sucrose, although it was abolished by the addition of D­fructose. S. mutans biofilm formation was significantly inhibited by growth in sucrose media supplemented with 1 and 4% D­tagatose compared with that in a culture containing sucrose alone, while S. mutans formed granular biofilms in the presence of this rare sugar. The inhibitory effect of D­tagatose on S. mutans biofilm formation was significantly more evident than that of xylitol. Growth in sucrose media supplemented with D­tagatose significantly decreased the expression of glucosyltransferase, exo­ß­fructosidase and D­fructose­specific phosphotransferase genes but not the expression of fructosyltransferase compared with the culture containing sucrose only. The activity of cell­associated glucosyltransferase in S. mutans was inhibited by 4% D­tagatose. These results indicate that D­tagatose reduces water­insoluble glucan production from sucrose by inhibiting glucosyltransferase activities, which limits access to the free D­fructose released during this process and retards the growth of S. mutans. Therefore, foods and oral care products containing D­tagatose are anticipated to reduce the risk of caries by inhibiting S. mutans biofilm formation.


Assuntos
Biofilmes/efeitos dos fármacos , Hexoses/farmacologia , Streptococcus mutans/efeitos dos fármacos , Streptococcus mutans/crescimento & desenvolvimento , Metabolismo dos Carboidratos/efeitos dos fármacos , Metabolismo dos Carboidratos/genética , Regulação Bacteriana da Expressão Gênica/efeitos dos fármacos , Regulação Enzimológica da Expressão Gênica , Hexoses/metabolismo , Streptococcus mutans/genética , Streptococcus mutans/metabolismo
3.
Nat Commun ; 8(1): 508, 2017 09 11.
Artigo em Inglês | MEDLINE | ID: mdl-28894084

RESUMO

The skin surface area varies flexibly in response to body shape changes. Skin homeostasis is maintained by stem cells residing in the basal layer of the interfollicular epidermis. However, how the interfollicular epidermal stem cells response to physiological body shape changes remains elusive. Here, we identify a highly proliferative interfollicular epidermal basal cell population in the rapidly expanding abdominal skin of pregnant mice. These cells express Tbx3 that is necessary for their propagation to drive skin expansion. The Tbx3+ basal cells are generated from Axin2+ interfollicular epidermal stem cells through planar-oriented asymmetric or symmetric cell divisions, and express transit-amplifying cell marker CD71. This biased division of Axin2+ interfollicular epidermal stem cells is induced by Sfrp1 and Igfbp2 proteins secreted from dermal cells. The Tbx3+ basal cells promote wound repair, which is enhanced by Sfrp1 and Igfbp2. This study elucidates the interfollicular epidermal stem cell/progeny organisation during pregnancy and suggests its application in regenerative medicine.The abdominal skin expands rapidly during pregnancy. Here the authors show that a population of highly proliferative stem cell progenies expressing the transcription factor Tbx3 is required for abdominal skin expansion in pregnant mice.


Assuntos
Derme/metabolismo , Células Epiteliais/metabolismo , Gravidez/metabolismo , Células-Tronco/citologia , Proteínas com Domínio T/metabolismo , Animais , Proteína Axina/genética , Proteína Axina/metabolismo , Proliferação de Células , Derme/citologia , Derme/crescimento & desenvolvimento , Células Epiteliais/citologia , Feminino , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/genética , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Masculino , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Camundongos , Gravidez/genética , Regeneração , Pele/citologia , Pele/crescimento & desenvolvimento , Pele/metabolismo , Células-Tronco/metabolismo , Proteínas com Domínio T/genética
4.
Ecotoxicol Environ Saf ; 144: 338-350, 2017 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-28646739

RESUMO

Water samples were collected from effluent-dominated urban streams in Tokushima, Kyoto, and Saitama in Japan to roughly determine the contribution of pharmaceuticals and personal care products (PPCPs) and surfactants to whole toxicity of the water. Approximately 100 PPCPs including anionic surfactants such as linear alkylbenzene sulfonate (LAS), were chemically analyzed. Using 14 water samples, chronic or sub-chronic toxicity tests were conducted on three aquatic species, the green alga Raphidocelis subcapitata, the cladoceran Ceriodaphnia dubia, and the zebrafish Danio rerio. Bioassays for the selected individual PPCPs were conducted using the three species. Assuming the concentration addition (CA) model, the contribution of each PPCP to the whole toxicity of the riverwater was estimated based on toxicity unit (TU). The contribution of PPCPs, which primarily consists of a few antibiotic agents such as triclosan and clarithromycin, ranged from 0.9% to 69% of the whole toxicity of the water samples for algae, whereas the selected LAS congeners accounted for at most 5.3%. In contrast, the contribution of LAS ranged from 0.067% to 86% and from 0.021% to 27% of the whole toxicity for cladoceran and zebrafish, respectively, whereas that of PPCPs for these species was at most 2.1% at all sampling points. Our results suggest a limited contribution of PPCPs except for antimicrobial agents and the possible substantial contribution of LAS to toxicity in cladocerans and zebrafish.


Assuntos
Cosméticos/toxicidade , Preparações Farmacêuticas/análise , Rios/química , Tensoativos/toxicidade , Poluentes Químicos da Água/toxicidade , Animais , Bioensaio , Clorófitas/efeitos dos fármacos , Cosméticos/análise , Daphnia/efeitos dos fármacos , Embrião não Mamífero/efeitos dos fármacos , Japão , Tensoativos/análise , Testes de Toxicidade Crônica , Urbanização , Poluentes Químicos da Água/análise , Peixe-Zebra/embriologia
5.
Sci Rep ; 7: 43136, 2017 02 23.
Artigo em Inglês | MEDLINE | ID: mdl-28230102

RESUMO

Enterohemorrhagic Escherichia coli (EHEC) O26 infections cause severe human diseases such as hemolytic uremic syndrome and encephalopathy, and is the predominant serogroup among non-O157 EHEC in many countries. Shiga toxin (Stx), which consists of two distinct types (Stx1 and Stx2), plays a central role in EHEC pathogenesis. The major stx gene type in EHEC O26 strains is stx1, although isolates with only stx2 have emerged in Japan since 2012 and have been reported in Europe. In this study, we selected 27 EHEC O26 strains isolated in Japan and identified a distinct genetic clade within sequence type (ST) 29, designated ST29C1, that carried only stx2 and had the plasmid gene profile ehxA+/katP-/espP+/etpD-. We showed that ST29C1 strains produced higher Stx2a levels, and greater virulence in Vero cells and in germ-free mice than other lineages. We also showed that ST29C1 was a distinct phylogenetic clade by SNP analysis using whole genome sequences and clearly differed from the major European EHEC O26 virulent clone, which was designated ST29C2 in this study. The combination of toxin production analysis, virulence analysis in Vero cells and germ-free mice, and phylogenetic analysis identified a newly emerging virulent EHEC clade.


Assuntos
Escherichia coli Êntero-Hemorrágica/classificação , Escherichia coli Êntero-Hemorrágica/genética , Infecções por Escherichia coli/microbiologia , Genótipo , Animais , Chlorocebus aethiops , Escherichia coli Êntero-Hemorrágica/crescimento & desenvolvimento , Humanos , Japão , Camundongos , Tipagem de Sequências Multilocus , Filogenia , Plasmídeos/análise , Polimorfismo de Nucleotídeo Único , Toxina Shiga/genética , Células Vero , Virulência , Sequenciamento Completo do Genoma
6.
Anaerobe ; 42: 81-88, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-27639596

RESUMO

Markerless gene deletion is necessary for multiple gene disruptions due to the limited number of antibiotic resistant markers for some bacteria. However, even in transformable strains, obtaining the expected mutation without a marker requires laborious screening of a large number of colonies. Previous studies had success in various bacteria with a counter-selection system where a conditional lethal gene was incorporated into the vector. We examined the efficacy of the mutated pheS gene (pheS*) as a counter-selective marker for gene deletion in Bacteroides. This mutation produces an amino acid substitution (A303G) in the alpha subunit of Bacteroides phenylalanyl tRNA synthetase, which in E. coli alters the specificity of the tRNA synthetase resulting in a conditional lethal mutation due to the incorporation of p-chloro-phenylalanine (p-Cl-Phe) into protein. B. fragilis YCH46 and B. thetaiotaomicron VPI-5482 transformed with a pheS*-harboring shuttle vector were clearly growth-inhibited in the presence of >5 mM p-Cl-Phe in liquid defined minimal media (DMM) and on DMM agar plates. A targeting plasmid was constructed to delete the genetic region for capsular polysaccharide PS2 in B. fragilis or PS1 in B. thetaiotaomicron. After counterselection, p-Cl-Phe-resistant colonies were generated at a frequency of 8.1 × 10-3 for B. fragilis and 1.7 × 10-3 for B. thetaiotaomicron. Of the p-Cl-Phe-resistant colonies, 4.2% and 72% harbored the correct genetic deletion for B. fragilis and B. thetaiotaomicron, respectively. These results indicate that mutated pheS is a useful counter-selective gene to construct markerless genetic deletions in Bacteroides.


Assuntos
Proteínas de Bactérias/genética , Bacteroides/genética , Deleção de Genes , Mutação , Fenilalanina-tRNA Ligase/genética , Subunidades Proteicas/genética , Sequência de Aminoácidos , Substituição de Aminoácidos , Cápsulas Bacterianas/metabolismo , Proteínas de Bactérias/metabolismo , Bacteroides/metabolismo , Meios de Cultura/farmacologia , Escherichia coli/genética , Escherichia coli/metabolismo , Fenclonina/farmacologia , Expressão Gênica , Genes Letais , Engenharia Genética , Marcadores Genéticos , Vetores Genéticos/química , Vetores Genéticos/metabolismo , Fenilalanina-tRNA Ligase/metabolismo , Polissacarídeos Bacterianos/metabolismo , Subunidades Proteicas/metabolismo , Alinhamento de Sequência
7.
Arch Oral Biol ; 68: 73-82, 2016 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-27107380

RESUMO

OBJECTIVE: Streptococcus mutans, a gram-positive oral bacterium, has been identified as one of the principal etiological agents of human dental caries. To clarify the nature of the difference anti-biofilm effect against S. mutans between Assam tea from Camellia sinensis var. assamica, partially fermented, and green tea from Camellia sinensis, non-fermented, active agents from the teas were purified. METHODS: Effects of Assam tea and green tea samples on biofilm were assessed by using the conventional titer plate method and the human saliva-coated hydroxyapatite discs. The purification and identification of inhibitors were performed by using ultrafiltration with centrifugal filter devices and high performance liquid chromatography. RESULTS: Assam tea has stronger biofilm inhibition activity against S. mutans than green tea. A substance of <10kDa in mass in Assam tea had a high concentration of galloylated catechins and a stronger biofilm inhibiting activity than green tea. In contrast, substances >10kDa in mass from green tea included higher concentrations of polysaccharides composed of galacturonic acid, such as pectin, that enhance biofilm formation. CONCLUSIONS: The higher concentrations of galloylated catechins in Assam tea may assist in prevention of dental caries, whereas in green tea, this mode of inhibition was likely offset by the presence of pectin. Purification of catechins in partially fermented Assam tea with lower-molecular-weight polysaccharide than pectin may be useful for developing oral care products such as toothpaste and oral care gel pastes.


Assuntos
Biofilmes/efeitos dos fármacos , Camellia sinensis/química , Extratos Vegetais/farmacologia , Streptococcus mutans/efeitos dos fármacos , Streptococcus mutans/fisiologia , Chá/química , Biofilmes/crescimento & desenvolvimento , Catequina/farmacologia , Cárie Dentária/microbiologia , Cárie Dentária/prevenção & controle , Durapatita , Ácidos Hexurônicos/farmacologia , Humanos , Pectinas/farmacologia , Saliva/microbiologia , Cremes Dentais/química
8.
Springerplus ; 5: 244, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27027597

RESUMO

Physical fitness is an indicator of systemic well-being in humans. Little is known about the role of physical fitness for maintaining systemic health in the elderly. Here, we study elderly subjects to determine the relationships between physical fitness and CD56 and CD16 surface NK cell markers on peripheral blood lymphocytes, as well as to analyze the relationship between the surface markers and incidence of death. We selected 253 independent elderly subjects (122 female; 131 male) who were 79-80 years old. Subjects having a higher proportion of CD56(dim)CD16(high) within CD56(+)CD16(+) cells, or ration of CD56(dim)CD16(high) and CD56(dim)CD16(-) cells had a significant positive correlation with maximum bilateral knee extensor strength/weight (kg) (r = 0.425; P < 0.0001 or r = 0.323; P < 0.0001). In contrast, an increased proportion of CD56(bright)CD16(-) cells within lymphocyte significantly negatively correlated with the maximum bilateral knee extensor strength/weight (kg) (r = -0.290; P = 0.0004); and these subjects had a significantly lower mortality during the 5 years following measurement of death. Therefore, we found that a synergistic effect of the right and left leg muscle strength was associated with proportion of matured NK and NKT cells and induced a low proportion of CD56(bright)CD16(-) cells within lymphocyte. Moreover, the low proportion of CD56(bright)CD16(-) cells was associated with incidence of death. In conclusion, measurements of physical fitness, the proportion of CD56(dim)CD16(high) within CD56(+)CD16(+) cells, the ratio of CD56(dim)CD56(high) and CD56(dim)CD16(-) cells, and the proportion of CD56(bright)C16(-) cells in lymphocytes are important indicators to check elderly health.

9.
BMC Oral Health ; 16: 40, 2016 Mar 22.
Artigo em Inglês | MEDLINE | ID: mdl-27001253

RESUMO

BACKGROUND: Dental plaque formed on tooth surfaces is a complex ecosystem composed of diverse oral bacteria and salivary components. Accumulation of dental plaque is a risk factor for dental caries and periodontal diseases. L-arginine has been reported to decrease the risk for dental caries by elevating plaque pH through the activity of arginine deiminase in oral bacteria. Here we evaluated the potential of L-arginine to remove established oral biofilms. METHODS: Biofilms were formed using human saliva mixed with Brain Heart Infusion broth supplemented with 1 % sucrose in multi-well plates or on plastic discs. After washing the biofilms with saline, citrate (10 mM, pH3.5), or L-arginine (0.5 M, pH3.5), the retained biofilms were analyzed by crystal violet staining, scanning electron microscopy, and Illumina-based 16S rDNA sequencing. RESULTS: Washing with acidic L-arginine detached oral biofilms more efficiently than saline and significantly reduced biofilm mass retained in multi-well plates or on plastic discs. Illumina-based microbiota analysis showed that citrate (pH3.5) preferentially washed out Streptococcus from mature oral biofilm, whereas acidic L-arginine prepared with 10 mM citrate buffer (pH3.5) non-specifically removed microbial components of the oral biofilm. CONCLUSIONS: Acidic L-arginine prepared with citrate buffer (pH3.5) effectively destabilized and removed mature oral biofilms. The acidic L-arginine solution described here could be used as an additive that enhances the efficacy of mouth rinses used in oral hygiene.


Assuntos
Arginina/farmacologia , Biofilmes/efeitos dos fármacos , Placa Dentária/prevenção & controle , Cárie Dentária/prevenção & controle , Humanos , Saliva
10.
PLoS One ; 11(2): e0148887, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-26859882

RESUMO

Phase changes in Bacteroides fragilis, a member of the human colonic microbiota, mediate variations in a vast array of cell surface molecules, such as capsular polysaccharides and outer membrane proteins through DNA inversion. The results of the present study show that outer membrane vesicle (OMV) formation in this anaerobe is also controlled by DNA inversions at two distantly localized promoters, IVp-I and IVp-II that are associated with extracellular polysaccharide biosynthesis and the expression of outer membrane proteins. These promoter inversions are mediated by a single tyrosine recombinase encoded by BF2766 (orthologous to tsr19 in strain NCTC9343) in B. fragilis YCH46, which is located near IVp-I. A series of BF2766 mutants were constructed in which the two promoters were locked in different configurations (IVp-I/IVp-II = ON/ON, OFF/OFF, ON/OFF or OFF/ON). ON/ON B. fragilis mutants exhibited hypervesiculating, whereas the other mutants formed only a trace amount of OMVs. The hypervesiculating ON/ON mutants showed higher resistance to treatment with bile, LL-37, and human ß-defensin 2. Incubation of wild-type cells with 5% bile increased the population of cells with the ON/ON genotype. These results indicate that B. fragilis regulates the formation of OMVs through DNA inversions at two distantly related promoter regions in response to membrane stress, although the mechanism underlying the interplay between the two regions controlled by the invertible promoters remains unknown.


Assuntos
Proteínas da Membrana Bacteriana Externa/biossíntese , Bacteroides fragilis/metabolismo , Inversão de Sequência/genética , Antibacterianos/farmacologia , Bacteroides fragilis/efeitos dos fármacos , Bacteroides fragilis/fisiologia , Membrana Celular/metabolismo , Farmacorresistência Bacteriana , Vesículas Extracelulares/metabolismo , Vesículas Extracelulares/fisiologia , Regulação Bacteriana da Expressão Gênica/genética , Regulação Bacteriana da Expressão Gênica/fisiologia , Regiões Promotoras Genéticas/genética , Regiões Promotoras Genéticas/fisiologia , Proteômica , Inversão de Sequência/fisiologia
11.
Behav Brain Res ; 292: 184-93, 2015 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-26057360

RESUMO

Many patients who have received chemotherapy to treat cancer experience depressive- and anxiety-like symptoms or cognitive impairment. However, despite the evidence for this, the underlying mechanisms are still not understood. This study investigated behavioral and biochemical changes upon treatment with doxorubicin and cyclophosphamide, focusing on mental and cognitive systems, as well as neurogenesis in male rats. Doxorubicin (2 mg/kg), cyclophosphamide (50 mg/kg), and the combination of doxorubicin and cyclophosphamide were injected intraperitoneally once per week for 4 weeks. In particular, the co-administration of doxorubicin and cyclophosphamide produced anhedonia-like, anxiety-like, and spatial cognitive impairments in rats. It also reduced both the number of proliferating cells in the subgranular zone of the hippocampal dentate gyrus and their survival. Serum brain-derived neurotrophic factor (BDNF) levels were decreased along with chemotherapy-induced decreases in platelet levels. However, hippocampal BDNF levels and Bdnf mRNA levels were not decreased by this treatment. On the other hand, hippocampal cyclin D1 levels were significantly decreased by chemotherapy. These results suggest that the co-administration of doxorubicin and cyclophosphamide induces psychological and cognitive impairment, in addition to negatively affecting hippocampal neurogenesis, which may be related to hippocampal cyclin D1 levels, but not hippocampal BDNF levels.


Assuntos
Ansiedade/induzido quimicamente , Fator Neurotrófico Derivado do Encéfalo/metabolismo , Ciclina D1/metabolismo , Ciclofosfamida/farmacologia , Doxorrubicina/farmacologia , Hipocampo/efeitos dos fármacos , Neurogênese/efeitos dos fármacos , Animais , Ansiedade/psicologia , Cognição/efeitos dos fármacos , Cognição/fisiologia , Transtornos Cognitivos/induzido quimicamente , Transtornos Cognitivos/psicologia , Depressão/psicologia , Masculino , Neurogênese/fisiologia , Ratos , Ratos Wistar
12.
PLoS One ; 9(6): e99263, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24896093

RESUMO

Fusobacterium nucleatum is a ubiquitous member of the human oral flora and is associated with the development of periodontitis and a variety of other types of polymicrobial infections of the mucosa. In the oral cavity, this species is one of the few that is prevalent in both healthy and diseased subgingival plaque. Using microarray analysis, we examined the transcriptional response of F. nucleatum subspecies nucleatum to whole blood in order to identify some of the genetic responses that might occur during the transition from health to disease. From these studies, we identified a sialic acid catabolism operon that was induced by the presence of blood. We subsequently confirmed that this operon was inducible by the presence of synthetic sialic acid, but we found no evidence suggesting sialic acid was used as a major carbon source. However, this organism was found to possess a de novo synthesized surface sialylation ability that is widely conserved among the various F. nucleatum subspecies as well as in F. periodonticum. We provide evidence that fusobacterial sialylation does occur in the oral cavity irrespective of health status. Interestingly, only a minority of fusobacterial cells exhibit surface sialylation within dental plaque, whereas most cells are uniformly sialylated when grown in pure culture. The implications of these results are discussed.


Assuntos
Placa Dentária/microbiologia , Fusobactérias/metabolismo , Infecções por Fusobacterium/microbiologia , Boca/microbiologia , Ácido N-Acetilneuramínico/metabolismo , Periodontite/microbiologia , Western Blotting , Membrana Celular/metabolismo , Células Cultivadas , Placa Dentária/metabolismo , Fusobactérias/classificação , Fusobactérias/genética , Infecções por Fusobacterium/genética , Infecções por Fusobacterium/metabolismo , Humanos , Hibridização in Situ Fluorescente , Boca/metabolismo , Periodontite/metabolismo , RNA Bacteriano/genética , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Especificidade da Espécie
13.
J Appl Toxicol ; 33(11): 1222-9, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-22806922

RESUMO

Acute and chronic (or sub-chronic) toxicity of five selected antimicrobial agents, including triclosan (TCS), triclocarban (TCC), resorcinol, phenoxyethanol and p-thymol, was investigated using the conventional three-aquatic-organism battery. These compounds are widely used in cosmetics and other personal care products and their ecological risk has recently become a significant concern. As results of toxicity tests, TCS was found to be most strongly toxic for green algae [e.g. 72 h no observed effect concentration (NOEC) of 0.50 µg l(-1) ] among the selected compounds, followed by TCC, while TCC was more toxic or similar to TCS for Daphnia and fish (e.g. Daphnia 8 day NOEC of 1.9 µg l(-1) ). Having compared the predicted no effect concentration (PNEC) determined from the toxicity data with measured environmental concentrations (MEC), the preliminary ecological risk assessment of these five antimicrobials was conducted. The MEC/PNEC ratios of TCS and TCC were over 1 for some monitoring data, especially in urban streams with watershed areas without sewage service coverage, and their potential risk for green algae and Daphnia might be at a level of concern, although the contribution of TCS/TCC on the total toxicity of the those sites needs to be further investigated. For the three other antimicrobials, the maximum MEC/PNEC ratio for resorcinol was 0.1-1, but those for phenoxyethanol and p-thymol were <0.1 and their risk to aquatic organisms is limited, although the additive effects with TCS, TCC and other antimicrobial agents, such as parabens, need to be further examined in future studies.


Assuntos
Anti-Infecciosos/toxicidade , Clorófitas/efeitos dos fármacos , Ecotoxicologia , Poluentes Químicos da Água/toxicidade , Animais , Anti-Infecciosos/química , Carbanilidas/química , Carbanilidas/toxicidade , Clorófitas/crescimento & desenvolvimento , Daphnia/efeitos dos fármacos , Daphnia/crescimento & desenvolvimento , Etilenoglicóis/química , Etilenoglicóis/toxicidade , Estrutura Molecular , Reprodução/efeitos dos fármacos , Resorcinóis/química , Resorcinóis/toxicidade , Medição de Risco , Timol/química , Timol/toxicidade , Testes de Toxicidade Aguda , Testes de Toxicidade Crônica , Triclosan/química , Triclosan/toxicidade
14.
Appl Environ Microbiol ; 77(5): 1572-80, 2011 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-21239559

RESUMO

The oral microbial flora consists of many beneficial species of bacteria that are associated with a healthy condition and control the progression of oral disease. Cooperative interactions between oral streptococci and the pathogens play important roles in the development of dental biofilms in the oral cavity. To determine the roles of oral streptococci in multispecies biofilm development and the effects of the streptococci in biofilm formation, the active substances inhibiting Streptococcus mutans biofilm formation were purified from Streptococcus salivarius ATCC 9759 and HT9R culture supernatants using ion exchange and gel filtration chromatography. Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry analysis was performed, and the results were compared to databases. The S. salivarius HT9R genome sequence was determined and used to indentify candidate proteins for inhibition. The candidates inhibiting biofilms were identified as S. salivarius fructosyltransferase (FTF) and exo-beta-d-fructosidase (FruA). The activity of the inhibitors was elevated in the presence of sucrose, and the inhibitory effects were dependent on the sucrose concentration in the biofilm formation assay medium. Purified and commercial FruA from Aspergillus niger (31.6% identity and 59.6% similarity to the amino acid sequence of FruA from S. salivarius HT9R) completely inhibited S. mutans GS-5 biofilm formation on saliva-coated polystyrene and hydroxyapatite surfaces. Inhibition was induced by decreasing polysaccharide production, which is dependent on sucrose digestion rather than fructan digestion. The data indicate that S. salivarius produces large quantities of FruA and that FruA alone may play an important role in multispecies microbial interactions for sucrose-dependent biofilm formation in the oral cavity.


Assuntos
Antibacterianos/metabolismo , Proteínas de Bactérias/metabolismo , Biofilmes/efeitos dos fármacos , Glicosídeo Hidrolases/metabolismo , Hexosiltransferases/metabolismo , Streptococcus mutans/efeitos dos fármacos , Antibacterianos/química , Antibacterianos/isolamento & purificação , Aspergillus niger/enzimologia , Proteínas de Bactérias/química , Proteínas de Bactérias/isolamento & purificação , Biofilmes/crescimento & desenvolvimento , Cromatografia em Gel , Proteínas Fúngicas/isolamento & purificação , Proteínas Fúngicas/metabolismo , Glicosídeo Hidrolases/química , Glicosídeo Hidrolases/isolamento & purificação , Hexosiltransferases/química , Hexosiltransferases/isolamento & purificação , Níger , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Streptococcus mutans/crescimento & desenvolvimento , Sacarose/metabolismo
15.
Curr Microbiol ; 62(1): 1-6, 2011 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-20458481

RESUMO

We analysed fragments of the formyltetrahydrofolate synthetase (FTHFS) gene, which encodes a key enzyme in reductive acetogenesis, from the bacterial flora in the proximal (PC) and mid (MC) colon of three ostriches to assess and compare bacterial diversity in this organ. Two clone libraries of FTHFS fragments were constructed from DNA extracted from digesta of the PC and MC, and a total of 46 cloned sequences were analysed from each library. A wide variety of FTHFS sequences were recovered. The coverage of the PC and MC libraries was 90.0% and 83.3%, respectively. Shannon-Wiener index (H') and Chao1 of the MC library were higher than those of PC library. The sequences from each library were classified into 15 operational taxonomic units (OTUs) and clusters. Only four OTUs in cluster I were distantly related to known acetogens from human feces and rumen, suggesting the presence of the novel acetogens. Phylogenetic analysis suggests that composition of FTHFS sequences differs for the PC and MC.


Assuntos
Bactérias/classificação , Bactérias/genética , Colo/microbiologia , Formiato-Tetra-Hidrofolato Ligase/genética , Variação Genética , Struthioniformes/microbiologia , Animais , Bactérias/enzimologia , Proteínas de Bactérias/genética , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , Dados de Sequência Molecular , Filogenia , Análise de Sequência de DNA
16.
Clin Lab ; 56(11-12): 505-12, 2010.
Artigo em Inglês | MEDLINE | ID: mdl-21141433

RESUMO

BACKGROUND: In the elderly, the necessity to promote oral health is increasing to improve their quality of life. The prediction of dental caries risk makes it possible to prolong the life span of teeth. To develop a quick diagnosis system for caries risk, two methods, the modified Saliva Check SM and Saliva Check sIgA, were investigated in elderly patients. METHODS: We developed a caries risk quick assessment system using Saliva Check sIgA that specifically recognizes secretory IgA (sIgA) in saliva against the binding site of the mutans streptococci (MS) to the salivary-coated tooth surface; and combined it with a modified Saliva Check SM to determine the number of MS. One hundred eighty three patients (80 females, 103 males) who participated in 2005 (average age 77 years) and in 2006 (average age 78 years) were assessed for caries risk using the systems in this cohort study. RESULTS: Subjects with a positive Saliva Check sIgA showed a significantly lower increment of decayed and filled teeth number (DFT) on the coronal surface; whereas those testing negative had root decay and increased filled teeth numbers (RDFT) at the root surface during the following year. The combination of Saliva Check sIgA and modified Saliva Check SM showed the subjects with Saliva Check sIgA positive and modified Saliva Check SM negative had less than half of the increment of DFT than other groups. In the other groups, Saliva Check sIgA negative and modified Saliva Check SM positive detected >90% of the subjects with an MS level of >5 x 10(5)/mL of saliva in patients that were assessed in 2006. This suggests these subjects may need extensive care. CONCLUSIONS: This new combination system significantly evaluates the caries risk to predict future incidence for dental caries on the coronal surface and may be useful for risk diagnosis of caries during a visit to the dental office.


Assuntos
Cárie Dentária/diagnóstico , Idoso , Idoso de 80 Anos ou mais , Anticorpos Antibacterianos/análise , Estudos de Coortes , Cárie Dentária/imunologia , Cárie Dentária/microbiologia , Restauração Dentária Permanente , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Imunoglobulina A Secretora/análise , Masculino , Valor Preditivo dos Testes , Fatores de Risco , Saliva/química , Saliva/microbiologia , Estatísticas não Paramétricas , Streptococcus mutans/isolamento & purificação
17.
Exp Biol Med (Maywood) ; 234(12): 1525-36, 2009 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-19934373

RESUMO

The non-obese diabetic mouse (NOD) is the most characterized model used to study insulin-dependent type 1 diabetes mellitus (IDDM) and Sjoögren's syndrome (SS). In a previous report, we found NOD.E2f1(-/-) mice show a greater progressive development to IDDM and SS compared to NOD mice. Our previous data indicated a progressive decrease in regulatory T cells (CD4(+)CD25(+)) and a decrease in the systemic secretion systems for insulin, and saliva was associated with the progression of IDDM and SS. Therefore, to define the mechanism of early-onset IDDM SS in E2F-1 deficient NOD mice required further investigation by producing E2F-1 deficient NOD/SCID mice in which the T and B cells do not develop. The purpose here was to analyze the essential function of the E2F-1 molecule in the development of IDDM and SS; and the dysfunction of the pancreas islet and salivary gland in the NOD background using NOD/SCID mice. We produced NOD/SCID.E2f1(-/-) mice using homologous recombination; determined diabetes development; measured saliva and insulin production; and performed a histological analysis. The deficient mice showed a decreasing volume of saliva; no infiltration of lymphocytes into salivary glands; no development of diabetes; and no protein localization of FGFR-2b in the ducts of the salivary gland that regulates submandibular gland proliferation and morphogenesis. Therefore, we considered a deficiency in E2F-1 induces a decrease in regulatory T cells and an increase in auto-reactive T cells; however, the E2F-1 deficiency is not associated with T and B cells-independent dysfunction of pancreatic beta cell in insulin secretion. Further, the E2F-1 deficiency is associated with T and B cells-independent dysfunction of the salivary gland exhibits a decrease in saliva production volume. We suggest E2F-1 may be also associated with the differentiation of exocrine cells in the duct where FGFR-2b is expressed in the salivary gland. The E2F-1 deficient NOD/SCID mouse model is useful for showing the development of the salivary gland; and is also useful for various experiments in humanized mice.


Assuntos
Diabetes Mellitus Tipo 1/metabolismo , Fator de Transcrição E2F1 , Ilhotas Pancreáticas/metabolismo , Saliva/metabolismo , Glândulas Salivares/metabolismo , Síndrome de Sjogren/metabolismo , Animais , Linfócitos B , Diabetes Mellitus Tipo 1/genética , Diabetes Mellitus Tipo 1/patologia , Modelos Animais de Doenças , Insulina , Ilhotas Pancreáticas/patologia , Camundongos , Camundongos Endogâmicos NOD , Camundongos Knockout , Camundongos SCID , Receptor Tipo 2 de Fator de Crescimento de Fibroblastos/metabolismo , Glândulas Salivares/crescimento & desenvolvimento , Glândulas Salivares/patologia , Síndrome de Sjogren/genética , Síndrome de Sjogren/patologia , Linfócitos T Reguladores
18.
Jpn J Infect Dis ; 60(1): 23-8, 2007 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-17314421

RESUMO

We evaluated the effects of special oral care using a toothbrush with combined irrigation and suctioning functions, along with povidone-iodine to treat oral bacteria and mucositis, in esophageal cancer patients undergoing chemoradiotherapy. In the special care group, oral hygiene was performed 3 days a week after dinner. Bacteria in saliva and plague samples were measured at various sampling points after chemoradiotherapy. The incidence of mucositis was significantly reduced in the special care group in comparison with the control group. Total streptococci were significantly decreased in the opportunistic pathogens-positive and lower-level mutans streptococci control group during chemoradiotherapy, but they were not reduced in the opportunistic pathogens-negative and higher-level mutans streptococci control groups or in the special care group. Our results showed that a special oral care regimen enabled the total population of streptococci microflora to remain stable, was negatively correlated with opportunistic pathogens and positively correlated with mutans streptococci infection, and prevented the development of mucositis.


Assuntos
Carcinoma de Células Escamosas/microbiologia , Neoplasias Esofágicas/microbiologia , Higiene Bucal/métodos , Estomatite/microbiologia , Idoso , Idoso de 80 Anos ou mais , Carcinoma de Células Escamosas/tratamento farmacológico , Carcinoma de Células Escamosas/radioterapia , Terapia Combinada , Neoplasias Esofágicas/tratamento farmacológico , Neoplasias Esofágicas/radioterapia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Povidona-Iodo/administração & dosagem , Estomatite/tratamento farmacológico , Estomatite/prevenção & controle , Infecções Estreptocócicas/tratamento farmacológico , Infecções Estreptocócicas/prevenção & controle , Streptococcus/crescimento & desenvolvimento , Streptococcus/isolamento & purificação
19.
FEMS Immunol Med Microbiol ; 49(1): 84-90, 2007 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-17266714

RESUMO

The amino acid residues 361-386 of Streptococcus mutans PAc includes an important region associated with the interaction between S. mutans and salivary components. We investigated the relationships between levels of the anti-PAc (361-386) peptide antibody (PPA) in saliva and periodontal status in 281 elderly subjects (mean age 77 years; 118 females, 163 males) by assessing dental calculus (CA), attachment loss (AL), pocket depth (PD), bleeding on probing (BOP) and various blood parameters. Enzyme-linked immunosorbent assay results revealed that subjects with a PPA level of greater than 0.1 (PPA detected group) showed a lower average value for number of sites with more than 6 mm of AL/6 points x 100/tooth (rAL6) than those with a PPA level of less than 0.1 (PPA not detected group). Furthermore, average values for rAL6 were significantly lower in the PPA detected group, and BOP, AL and rAL6 correlated positively and significantly with the percentage of eosinophils present in leukocytes in female subjects in both groups. PPA level had a negative correlation with percentages of basophils and eosinophils. The results indicate that systemic increases in numbers of eosinophils and basophils are associated with the development of periodontal diseases, while PPA level may be a useful indicator of periodontal status.


Assuntos
Proteínas de Bactérias/imunologia , Basófilos/imunologia , Eosinófilos/imunologia , Imunoglobulina A/imunologia , Glicoproteínas de Membrana/imunologia , Fragmentos de Peptídeos/imunologia , Doenças Periodontais/imunologia , Streptococcus mutans/imunologia , Idoso , Sequência de Aminoácidos , Feminino , Humanos , Imunoglobulina A/biossíntese , Estudos Longitudinais , Masculino , Dados de Sequência Molecular , Doenças Periodontais/etiologia , Doenças Periodontais/microbiologia , Índice Periodontal , Saliva/imunologia
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