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1.
J Appl Microbiol ; 133(3): 1341-1352, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-35603698

RESUMO

AIMS: We present a dynamic typodont biofilm model (DTBM) incorporating (1) human dentition anatomy, (2) fluid flow over intermittently fluid bathed tooth surfaces and (3) an oxic headspace to allow aerobic and anaerobic niches to develop naturally, as a screening tool to assess the effect of stannous fluoride (SnF2 ) toothpaste against a simulated human plaque biofilm (SPB). METHODS AND RESULTS: First, hydroxyapatite (HA) coupons were inoculated with human saliva/plaque and cultured at 37°C under air. Selected species representative of common commensal and anaerobic pathogens were quantified for relative abundance changes over 4 days by PCR densitometry to confirm the culture conditions allowed the proliferation of these species. A continuous culture DTBM reactor on a rocker table was inoculated with saliva/plaque and incubated at 37°C for 24 h. Tooth shear stress was estimated by particle tracking. A SnF2 toothpaste solution, or a sham rise was administered twice daily for 3 days to mimic routine oral hygiene. SPB biomass was assessed by total bacterial DNA and methylene blue (MB) staining. Early colonizer aerobes and late colonizer anaerobes species were detected in the HA and DTBM, and the trends in changing abundance were consistent with those seen clinically. CONCLUSIONS: Treatment with the SnF2 solution showed significant reductions of 53.05% and 54.4% in the SPB by MB staining and DNA, respectively. SIGNIFICANCE AND IMPACT OF STUDY: The model has potential for assessing dentition anatomy and fluid flow on the efficacy of antimicrobial efficacy against localized SPB and may be amenable to the plaque index clinical evaluation.


Assuntos
Fluoretos de Estanho , Cremes Dentais , Biofilmes , Humanos , Saliva , Fluoretos de Estanho/uso terapêutico , Cremes Dentais/farmacologia , Cremes Dentais/uso terapêutico
3.
J Cosmet Laser Ther ; 22(4-5): 200-204, 2020 Jul 03.
Artigo em Inglês | MEDLINE | ID: mdl-33059510

RESUMO

This study was performed to investigate the user perception of a cordless, motorized electronically controlled delivery system for botulinum toxin type A. Forty-six post-graduate students of varying experience levels of botulinum toxin injections and four members of the faculty from the MSc programme in Esthetic Medicine at Queen Mary University London participated in a demonstration of a motorized injection device. Thereafter, they performed a number of injections on a high fidelity silicone simulation model before completing a nine-item questionnaire. The more experienced injectors tended to appreciate the accuracy of the device more than the less experienced participants. Seventy-eight percent of participants said the device improved accuracy, particularly when administering small doses. Eighty-four percent leaned toward a favorable general view of the device. Forty-seven percent would possibly consider purchasing the device. Sixty-one percent would consider recommending the device to a colleague. The main advantage of the motorized injection device was the improved accuracy enabling delivery of small and precise doses. This may open up the possibility of new approaches to botulinum toxin treatments.


Assuntos
Atitude do Pessoal de Saúde , Toxinas Botulínicas Tipo A/administração & dosagem , Injeções/instrumentação , Fármacos Neuromusculares/administração & dosagem , Humanos , Projetos Piloto , Tecnologia
4.
Ann Intern Med ; 173(12): 989-1001, 2020 12 15.
Artigo em Inglês | MEDLINE | ID: mdl-32894695

RESUMO

DESCRIPTION: Hereditary hemorrhagic telangiectasia (HHT) is an autosomal dominant disease with an estimated prevalence of 1 in 5000 that is characterized by the presence of vascular malformations (VMs). These result in chronic bleeding, acute hemorrhage, and complications from shunting through VMs. The goal of the Second International HHT Guidelines process was to develop evidence-based consensus guidelines for the management and prevention of HHT-related symptoms and complications. METHODS: The guidelines were developed using the AGREE II (Appraisal of Guidelines for Research and Evaluation II) framework and GRADE (Grading of Recommendations Assessment, Development and Evaluation) methodology. The guidelines expert panel included expert physicians (clinical and genetic) in HHT from 15 countries, guidelines methodologists, health care workers, health care administrators, patient advocacy representatives, and persons with HHT. During the preconference process, the expert panel generated clinically relevant questions in 6 priority topic areas. A systematic literature search was done in June 2019, and articles meeting a priori criteria were included to generate evidence tables, which were used as the basis for recommendation development. The expert panel subsequently convened during a guidelines conference to conduct a structured consensus process, during which recommendations reaching at least 80% consensus were discussed and approved. RECOMMENDATIONS: The expert panel generated and approved 6 new recommendations for each of the following 6 priority topic areas: epistaxis, gastrointestinal bleeding, anemia and iron deficiency, liver VMs, pediatric care, and pregnancy and delivery (36 total). The recommendations highlight new evidence in existing topics from the first International HHT Guidelines and provide guidance in 3 new areas: anemia, pediatrics, and pregnancy and delivery. These recommendations should facilitate implementation of key components of HHT care into clinical practice.


Assuntos
Telangiectasia Hemorrágica Hereditária/diagnóstico , Telangiectasia Hemorrágica Hereditária/terapia , Anemia/etiologia , Anemia/terapia , Malformações Arteriovenosas/etiologia , Malformações Arteriovenosas/terapia , Criança , Epistaxe/etiologia , Epistaxe/terapia , Hemorragia Gastrointestinal/etiologia , Hemorragia Gastrointestinal/terapia , Doenças Genéticas Inatas/etiologia , Doenças Genéticas Inatas/terapia , Humanos , Fígado/irrigação sanguínea , Telangiectasia Hemorrágica Hereditária/complicações
6.
BMC Oral Health ; 20(1): 230, 2020 08 21.
Artigo em Inglês | MEDLINE | ID: mdl-32825831

RESUMO

BACKGROUND: Dental plaque biofilms are the causative agents of caries, gingivitis and periodontitis. Both mechanical and chemical strategies are used in routine oral hygiene strategies to reduce plaque build-up. If allowed to mature biofilms can create anoxic microenvironments leading to communities which harbor pathogenic Gram-negative anaerobes. When subjected to high velocity fluid jets and sprays biofilms can be fluidized which disrupts the biofilm structure and allows the more efficient delivery of antimicrobial agents. METHODS: To investigate how such jets may disrupt anoxic niches in the biofilm, we used planar optodes to measure the dissolved oxygen (DO) concentration at the base of in-vitro biofilms grown from human saliva and dental plaque. These biofilms were subject to "shooting" treatments with a commercial high velocity microspray (HVM) device. RESULTS: HVM treatment resulted in removal of much of the biofilm and a concurrent rapid shift from anoxic to oxic conditions at the base of the surrounding biofilm. We also assessed the impact of HVM treatment on the microbial community by tracking 7 target species by qPCR. There was a general reduction in copy numbers of the universal 16S RNA by approximately 95%, and changes of individual species in the target region ranged from approximately 1 to 4 log reductions. CONCLUSION: We concluded that high velocity microsprays removed a sufficient amount of biofilm to disrupt the anoxic region at the biofilm-surface interface.


Assuntos
Placa Dentária , Microbiota , Biofilmes , Humanos , Oxigênio , Saliva
7.
Mol Oral Microbiol ; 34(4): 131-152, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31034136

RESUMO

A comparative proteomic analysis was utilized to evaluate similarities and differences in membrane samples derived from the cariogenic bacterium Streptococcus mutans, including the wild-type strain and four mutants devoid of protein translocation machinery components, specifically ∆ffh, ∆yidC1, ∆yidC2, or ∆ffh/yidC1. The purpose of this work was to determine the extent to which the encoded proteins operate individually or in concert with one another and to identify the potential substrates of the respective pathways. Ffh is the principal protein component of the signal recognition particle (SRP), while yidC1 and yidC2 are dual paralogs encoding members of the YidC/Oxa/Alb family of membrane-localized chaperone insertases. Our results suggest that the co-translational SRP pathway works in concert with either YidC1 or YidC2 specifically, or with no preference for paralog, in the insertion of most membrane-localized substrates. A few instances were identified in which the SRP pathway alone, or one of the YidCs alone, appeared to be most relevant. These data shed light on underlying reasons for differing phenotypic consequences of ffh, yidC1 or yidC2 deletion. Our data further suggest that many membrane proteins present in a ∆yidC2 background may be non-functional, that ∆yidC1 is better able to adapt physiologically to the loss of this paralog, that shared phenotypic properties of ∆ffh and ∆yidC2 mutants can stem from impacts on different proteins, and that independent binding to ribosomal proteins is not a primary functional activity of YidC2. Lastly, genomic mutations accumulate in a ∆yidC2 background coincident with phenotypic reversion, including an apparent W138R suppressor mutation within yidC1.


Assuntos
Proteínas de Bactérias , Streptococcus mutans , Proteínas de Bactérias/genética , Proteínas de Bactérias/fisiologia , Proteínas de Membrana/genética , Proteínas de Membrana/fisiologia , Chaperonas Moleculares , Mutação , Proteômica , Partícula de Reconhecimento de Sinal , Streptococcus mutans/genética , Streptococcus mutans/metabolismo
8.
J Bacteriol ; 201(1)2019 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-30322852

RESUMO

Proper envelope biogenesis of Streptococcus mutans, a biofilm-forming and dental caries-causing oral pathogen, requires two paralogs (yidC1 and yidC2) of the universally conserved YidC/Oxa1/Alb3 family of membrane integral chaperones and insertases. The deletion of either paralog attenuates virulence in vivo, but the mechanisms of disruption remain unclear. Here, we determined whether the deletion of yidC affects cell surface properties, extracellular glucan production, and/or the structural organization of the exopolysaccharide (EPS) matrix and biophysical properties of S. mutans biofilm. Compared to the wild type, the ΔyidC2 mutant lacked staining with fluorescent vancomycin at the division septum, while the ΔyidC1 mutant resembled the wild type. Additionally, the deletion of either yidC1 or yidC2 resulted in less insoluble glucan synthesis but produced more soluble glucans, especially at early and mid-exponential-growth phases. Alteration of glucan synthesis by both mutants yielded biofilms with less dry weight and insoluble EPS. In particular, the deletion of yidC2 resulted in a significant reduction in biofilm biomass and pronounced defects in the spatial organization of the EPS matrix, thus modifying the three-dimensional (3D) biofilm architecture. The defective biofilm harbored smaller bacterial clusters with high cell density and less surrounding EPS than those of the wild type, which was stiffer in compression yet more susceptible to removal by shear. Together, our results indicate that the elimination of either yidC paralog results in changes to the cell envelope and glucan production that ultimately disrupts biofilm development and EPS matrix structure/composition, thereby altering the physical properties of the biofilms and facilitating their removal. YidC proteins, therefore, represent potential therapeutic targets for cariogenic biofilm control.IMPORTANCE YidC proteins are membrane-localized chaperone insertases that are universally conserved in all bacteria and are traditionally studied in the context of membrane protein insertion and assembly. Both YidC paralogs of the cariogenic pathogen Streptococcus mutans are required for proper envelope biogenesis and full virulence, indicating that these proteins may also contribute to optimal biofilm formation in streptococci. Here, we show that the deletion of either yidC results in changes to the structure and physical properties of the EPS matrix produced by S. mutans, ultimately impairing optimal biofilm development, diminishing its mechanical stability, and facilitating its removal. Importantly, the universal conservation of bacterial yidC orthologs, combined with our findings, provide a rationale for YidC as a possible drug target for antibiofilm therapies.


Assuntos
Proteínas de Bactérias/metabolismo , Biofilmes/crescimento & desenvolvimento , Fenômenos Biofísicos , Parede Celular/metabolismo , Matriz Extracelular de Substâncias Poliméricas/metabolismo , Glucanos/metabolismo , Streptococcus mutans/enzimologia , Proteínas de Bactérias/genética , Matriz Extracelular de Substâncias Poliméricas/química , Deleção de Genes , Glucanos/química , Streptococcus mutans/genética , Streptococcus mutans/crescimento & desenvolvimento
9.
Environ Microbiol Rep ; 10(1): 12-22, 2018 02.
Artigo em Inglês | MEDLINE | ID: mdl-29124888

RESUMO

Streptococcus mutans strongly influences the development of pathogenic biofilms associated with dental caries. Our understanding of S. mutans behaviour in biofilms is based on a few well-characterized laboratory strains; however, individual isolates vary widely in genome content and virulence-associated phenotypes, such as biofilm formation and environmental stress sensitivity. Using an ecological biofilm model, we assessed the impact of co-cultivation of several S. mutans isolates with Streptococcus oralis and Actinomyces naeslundii on biofilm composition following exposure to sucrose. The laboratory reference strain S. mutans UA159 and clinical isolates Smu44 (most aciduric), Smu56 (altered biofilm formation) and Smu81 (more sensitive to oxidative stress) were used. Our data revealed S. mutans isolates varied in their ability to compete and become dominant in the biofilm after the addition of sucrose, and this difference correlated with sensitivity to H2 O2 produced by S. oralis. Smu81 was particularly sensitive to H2 O2 and could not compete with S. oralis in mixed-species biofilm, despite forming robust biofilms on its own. Thus, diminished oxidative stress tolerance in S. mutans isolates can impair their ability to compete in complex biofilms, even in the presence of sucrose, which could influence the progression of a healthy biofilm community to one capable of causing disease.


Assuntos
Biofilmes/crescimento & desenvolvimento , Cárie Dentária/microbiologia , Interações Microbianas , Estresse Oxidativo/fisiologia , Streptococcus mutans/fisiologia , Actinomyces/fisiologia , Regulação Bacteriana da Expressão Gênica/efeitos dos fármacos , Peróxido de Hidrogênio/metabolismo , Peróxido de Hidrogênio/farmacologia , Concentração de Íons de Hidrogênio , Interações Microbianas/fisiologia , Complexos Multienzimáticos/genética , NADH NADPH Oxirredutases/genética , Streptococcus mutans/patogenicidade , Streptococcus oralis/fisiologia , Sacarose/metabolismo , Virulência/fisiologia
10.
mSystems ; 2(1)2017.
Artigo em Inglês | MEDLINE | ID: mdl-28066817

RESUMO

In the cariogenic Streptococcus mutans, competence development is regulated by the ComRS signaling system comprised of the ComR regulator and the ComS prepeptide to the competence signaling peptide XIP (ComX-inducing peptide). Aside from competence development, XIP signaling has been demonstrated to regulate cell lysis, and recently, the expression of bacteriocins, small antimicrobial peptides used by bacteria to inhibit closely related species. Our study further explores the effect of XIP signaling on the S. mutans transcriptome. RNA sequencing revealed that XIP induction resulted in a global change in gene expression that was consistent with a stress response. An increase in several membrane-bound regulators, including HdrRM and BrsRM, involved in bacteriocin production, and the VicRKX system, involved in acid tolerance and biofilm formation, was observed. Furthermore, global changes in gene expression corresponded to changes observed during the stringent response to amino acid starvation. Effects were also observed on genes involved in sugar transport and carbon catabolite repression and included the levQRST and levDEFG operons. Finally, our work identified a novel heat shock-responsive intergenic region, encoding a small RNA, with a potential role in competence shutoff. IMPORTANCE Genetic competence provides bacteria with an opportunity to increase genetic diversity or acquire novel traits conferring a survival advantage. In the cariogenic pathogen Streptococcus mutans, DNA transformation is regulated by the competence stimulating peptide XIP (ComX-inducing peptide). The present study utilizes high-throughput RNA sequencing (RNAseq) to provide a greater understanding of how global gene expression patterns change in response to XIP. Overall, our work demonstrates that in S. mutans, XIP signaling induces a response that resembles the stringent response to amino acid starvation. We further identify a novel heat shock-responsive intergenic region with a potential role in competence shutoff. Together, our results provide further evidence that multiple stress response mechanisms are linked through the genetic competence signaling pathway in S. mutans.

11.
Appl Environ Microbiol ; 82(7): 2187-201, 2016 Jan 29.
Artigo em Inglês | MEDLINE | ID: mdl-26826230

RESUMO

The ability of certain oral biofilm bacteria to moderate pH through arginine metabolism by the arginine deiminase system (ADS) is a deterrent to the development of dental caries. Here, we characterize a novel Streptococcus strain, designated strain A12, isolated from supragingival dental plaque of a caries-free individual. A12 not only expressed the ADS pathway at high levels under a variety of conditions but also effectively inhibited growth and two intercellular signaling pathways of the dental caries pathogen Streptococcus mutans. A12 produced copious amounts of H2O2 via the pyruvate oxidase enzyme that were sufficient to arrest the growth of S. mutans. A12 also produced a protease similar to challisin (Sgc) of Streptococcus gordonii that was able to block the competence-stimulating peptide (CSP)-ComDE signaling system, which is essential for bacteriocin production by S. mutans. Wild-type A12, but not an sgc mutant derivative, could protect the sensitive indicator strain Streptococcus sanguinis SK150 from killing by the bacteriocins of S. mutans. A12, but not S. gordonii, could also block the XIP (comX-inducing peptide) signaling pathway, which is the proximal regulator of genetic competence in S. mutans, but Sgc was not required for this activity. The complete genome sequence of A12 was determined, and phylogenomic analyses compared A12 to streptococcal reference genomes. A12 was most similar to Streptococcus australis and Streptococcus parasanguinis but sufficiently different that it may represent a new species. A12-like organisms may play crucial roles in the promotion of stable, health-associated oral biofilm communities by moderating plaque pH and interfering with the growth and virulence of caries pathogens.


Assuntos
Antibiose , Arginina/metabolismo , Placa Dentária/microbiologia , Streptococcus mutans/crescimento & desenvolvimento , Streptococcus/fisiologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Bacteriocinas/metabolismo , Bacteriocinas/farmacologia , Biofilmes , Regulação Bacteriana da Expressão Gênica , Filogenia , Streptococcus/classificação , Streptococcus/genética , Streptococcus/isolamento & purificação , Streptococcus mutans/efeitos dos fármacos , Streptococcus mutans/fisiologia
12.
Mol Microbiol ; 98(2): 302-17, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26172310

RESUMO

A unique 373 bp region (igr66) between grpE and dnaK of Streptococcus mutans lacks a promoter but is required for optimal production of DnaK. Northern blotting using probes specific to hrcA, igr66 or dnaK revealed multiple transcripts produced from the dnaK operon and 5'-RACE mapped 5' termini of multiple dnaK transcripts within igr66. One product mapped to a predicted 5'-SL (stem-loop) and two others mapped just 5' to Shine-Dalgarno (SD)-like sequences located immediately upstream to dnaK and to a predicted SL 120 bp upstream of the dnaK start codon (3'-SL). A collection of cat reporter-gene strains containing mutant derivatives of igr66 were engineered. Chloramphenicol acetyltransferase (CAT) activity varied greatly between strains, but there were no correlative changes in cat mRNA levels. Interestingly, mutations introduced into the SD-like sequences 5' to the 3'-SL resulted in an 83-98% decrease in CAT activity. Markerless point mutations introduced upstream of dnaK in the SD-like sequences impaired growth at elevated temperatures and resulted in up to a 40% decrease in DnaK protein after heat shock. Collectively, these results indicate processing within igr66 enhances translation in a temperature dependent manner via non-canonical ribosome binding sites positioned >120 bp upstream of dnaK.


Assuntos
Proteínas de Bactérias/genética , DNA Intergênico , Regulação Bacteriana da Expressão Gênica , Proteínas de Choque Térmico/genética , Processamento Pós-Transcricional do RNA , RNA Mensageiro/metabolismo , Streptococcus mutans/genética , Sequência de Aminoácidos , Sequência de Bases , Cloranfenicol O-Acetiltransferase/biossíntese , Proteínas de Ligação a DNA/genética , Proteínas de Choque Térmico HSP70/genética , Resposta ao Choque Térmico/genética , Mutação , Óperon , Regiões Promotoras Genéticas , RNA Mensageiro/genética , Proteínas Repressoras/genética
13.
Mol Microbiol ; 96(3): 463-82, 2015 May.
Artigo em Inglês | MEDLINE | ID: mdl-25620525

RESUMO

Streptococcus mutans displays complex regulation of genetic competence, with ComX controlling late competence gene transcription. The rcrRPQ operon has been shown to link oxidative stress tolerance, (p)ppGpp metabolism and competence in S. mutans. Importantly, an rcrR polar (ΔrcrR-P) mutant is hyper-transformable, but an rcrR non-polar (ΔrcrR-NP) mutant cannot be transformed. Transcriptome comparisons of the rcrR mutants using RNA-Seq and quantitative real-time polymerase chain reaction revealed little expression in the 5' region of comX in ΔrcrR-NP, but high level expression in the 3' region. Northern blotting with comX probes revealed two distinct transcripts in the ΔrcrR-P and ΔrcrR-NP strains, and 5' Rapid Amplification of cDNA Ends mapped the 5' terminus of the shorter transcript to nt +140 of the comX structural gene, where a unique 69-aa open reading frame, termed XrpA, was encoded in a different reading frame than ComX. Two single-nucleotide substitution mutants (comX::T162C; comX::T210A) were introduced to disrupt XrpA without affecting the sequence of ComX. When the mutations were in the ΔrcrR-NP genetic background, ComX production and transformation were restored. Overexpression of xrpA led to impaired growth in aerobic conditions and decreased transformability. These results reveal an unprecedented mechanism for competence regulation and stress tolerance by a gene product encoded within the comX gene that appears unique to S. mutans.


Assuntos
Competência de Transformação por DNA , Regulação Bacteriana da Expressão Gênica , Fases de Leitura Aberta , Estresse Oxidativo , Streptococcus mutans/genética , Streptococcus mutans/fisiologia , Perfilação da Expressão Gênica , Reação em Cadeia da Polimerase em Tempo Real , Análise de Sequência de RNA , Transcrição Gênica
14.
Genome Biol Evol ; 6(4): 741-53, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24625962

RESUMO

The genus Streptococcus comprises important pathogens that have a severe impact on human health and are responsible for substantial economic losses to agriculture. Here, we utilize 46 Streptococcus genome sequences (44 species), including eight species sequenced here, to provide the first genomic level insight into the evolutionary history and genetic basis underlying the functional diversity of all major groups of this genus. Gene gain/loss analysis revealed a dynamic pattern of genome evolution characterized by an initial period of gene gain followed by a period of loss, as the major groups within the genus diversified. This was followed by a period of genome expansion associated with the origins of the present extant species. The pattern is concordant with an emerging view that genomes evolve through a dynamic process of expansion and streamlining. A large proportion of the pan-genome has experienced lateral gene transfer (LGT) with causative factors, such as relatedness and shared environment, operating over different evolutionary scales. Multiple gene ontology terms were significantly enriched for each group, and mapping terms onto the phylogeny showed that those corresponding to genes born on branches leading to the major groups represented approximately one-fifth of those enriched. Furthermore, despite the extensive LGT, several biochemical characteristics have been retained since group formation, suggesting genomic cohesiveness through time, and that these characteristics may be fundamental to each group. For example, proteolysis: mitis group; urea metabolism: salivarius group; carbohydrate metabolism: pyogenic group; and transcription regulation: bovis group.


Assuntos
Evolução Molecular , Transferência Genética Horizontal/fisiologia , Genoma Bacteriano/fisiologia , Filogenia , Streptococcus/genética , Sequência de Bases , Humanos , Dados de Sequência Molecular
15.
Radiol Technol ; 84(6): 571-6, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23861517

RESUMO

Early in pregnancy, the female embryo's reproductive system begins developing from the müllerian ducts. When development fails to progress normally, various anomalies of the uterus, vagina, and urinary system can result. In some cases, these anomalies cause infertility and other reproductive problems. This literature review discusses the 6 classes of müllerian duct anomalies, the complications associated with each, and the imaging examinations used to diagnose them and further evaluate uterine anatomy. In addition, a case study of a class II anomaly, known as unicornuate uterus, is presented. As reproductive medicine advances and more women seek diagnosis of and treatment for fertility problems, radiologic technologists need to be aware of duct anomalies and their appearance with various imaging modalities, including ultrasonography, hysterosalpingography, and magnetic resonance imaging. In addition, technologists must be sensitive to the emotional needs of patients with müllerian duct anomalies.


Assuntos
Transtornos 46, XX do Desenvolvimento Sexual/diagnóstico por imagem , Transtornos 46, XX do Desenvolvimento Sexual/patologia , Anormalidades Congênitas/diagnóstico por imagem , Anormalidades Congênitas/patologia , Imageamento por Ressonância Magnética/métodos , Ductos Paramesonéfricos/anormalidades , Tomografia Computadorizada por Raios X/métodos , Adulto , Feminino , Humanos , Ductos Paramesonéfricos/diagnóstico por imagem , Ductos Paramesonéfricos/patologia
16.
PLoS One ; 8(4): e61358, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23613838

RESUMO

High coverage, whole genome shotgun (WGS) sequencing of 57 geographically- and genetically-diverse isolates of Streptococcus mutans from individuals of known dental caries status was recently completed. Of the 57 sequenced strains, fifteen isolates, were selected based primarily on differences in gene content and phenotypic characteristics known to affect virulence and compared with the reference strain UA159. A high degree of variability in these properties was observed between strains, with a broad spectrum of sensitivities to low pH, oxidative stress (air and paraquat) and exposure to competence stimulating peptide (CSP). Significant differences in autolytic behavior and in biofilm development in glucose or sucrose were also observed. Natural genetic competence varied among isolates, and this was correlated to the presence or absence of competence genes, comCDE and comX, and to bacteriocins. In general strains that lacked the ability to become competent possessed fewer genes for bacteriocins and immunity proteins or contained polymorphic variants of these genes. WGS sequence analysis of the pan-genome revealed, for the first time, components of a Type VII secretion system in several S. mutans strains, as well as two putative ORFs that encode possible collagen binding proteins located upstream of the cnm gene, which is associated with host cell invasiveness. The virulence of these particular strains was assessed in a wax-worm model. This is the first study to combine a comprehensive analysis of key virulence-related phenotypes with extensive genomic analysis of a pathogen that evolved closely with humans. Our analysis highlights the phenotypic diversity of S. mutans isolates and indicates that the species has evolved a variety of adaptive strategies to persist in the human oral cavity and, when conditions are favorable, to initiate disease.


Assuntos
Genoma Bacteriano/genética , Streptococcus mutans/genética , Proteínas de Bactérias/genética , Streptococcus mutans/classificação
17.
Artigo em Inglês | MEDLINE | ID: mdl-22995465

RESUMO

Plumbogummite PbAl(3)(PO(4))(2)(OH,H(2)O)(6) is a mineral of environmental significance and is a member of the alunite-jarosite supergroup. The molecular structure of the mineral has been investigated by Raman spectroscopy. The spectra of different plumbogummite specimens differ although there are many common features. The Raman spectra prove the spectral profile consisting of overlapping bands and shoulders. Raman bands and shoulders observed at 971, 980, 1002 and 1023 cm(-1) (China sample) and 913, 981, 996 and 1026 cm(-1) (Czech sample) are assigned to the ν(1) symmetric stretching modes of the (PO(4))(3-), at 1002 and 1023 cm(-1) (China) and 996 and 1026 cm(-1) to the ν(1) symmetric stretching vibrations of the (O(3)POH)(2-) units, and those at 1057, 1106 and 1182 (China) and at 1102, 1104 and 1179 cm(-1) (Czech) to the ν(3) (PO(4))(3-) and ν(3) (PO(3)) antisymmetric stretching vibrations. Raman bands and shoulders at 634, 613 and 579 cm(-1) (China) and 611 and 596 cm(-1) (Czech) are attributed to the ν(4) (δ) (PO(4))(3-) bending vibrations and those at 507, 494 and 464 cm(-1) (China) and 505 and 464 cm(-1) (Czech) to the ν(2) (δ) (PO(4))(3-) bending vibrations. The Raman spectrum of the OH stretching region is complex. Raman bands and shoulders are identified at 2824, 3121, 3249, 3372, 3479 and 3602 cm(-1) for plumbogummite from China, and at 3077, 3227, 3362, 3480, 3518 and 3601 cm(-1) for the Czech Republic sample. These bands are assigned to the ν OH stretching modes of water molecules and hydrogen ions. Approximate O-H⋯O hydrogen bond lengths inferred from the Raman spectra vary in the range >3.2-2.62Å (China) and >3.2-2.67Å (Czech). The minority presence of some carbonate ions in the plumbogummite (China sample) is connected with distinctive intensity increasing of the Raman band at 1106 cm(-1), in which may participate the ν(1) (CO(3))(2-) symmetric stretching vibration overlapped with phosphate stretching vibrations.


Assuntos
Chumbo/química , Minerais/química , Fosfatos/química , Análise Espectral Raman , Alumínio/química , Compostos de Alumínio/química , Compostos Férricos/química , Compostos de Potássio/química , Sulfatos/química
18.
Microbiology (Reading) ; 158(Pt 7): 1702-1712, 2012 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-22504439

RESUMO

The cariogenic bacterium Streptococcus mutans has two paralogues of the YidC/Oxa1/Alb3 family of membrane protein insertases/chaperones. Disruption of yidC2 results in loss of genetic competence, decreased membrane-associated ATPase activity and stress sensitivity (acid, osmotic and oxidative). Elimination of yidC1 has less severe effects, with little observable effect on growth or stress sensitivity. To examine the respective roles of YidC1 and YidC2, a conditional expression system was developed allowing simultaneous elimination of both endogenous YidCs. The function of the YidC C-terminal tails was also investigated and a chimeric YidC1 protein appended with the C terminus of YidC2 enabled YidC1 to complement a ΔyidC2 mutant for stress tolerance, ATP hydrolysis activity and extracellular glyceraldehyde-3-phosphate dehydrogenase (GAPDH) activity. Elimination of yidC1 or yidC2 affected levels of extracellular proteins, including GtfB, GtfC and adhesin P1 (AgI/II, PAc), which were increased without YidC1 but decreased in the absence of YidC2. Both yidC1 and yidC2 were shown to contribute to S. mutans biofilm formation and to cariogenicity in a rat model. Collectively, these results provide evidence that YidC1 and YidC2 contribute to cell surface biogenesis and protein secretion in S. mutans and that differences in stress sensitivity between the ΔyidC1 and ΔyidC2 mutants stem from a functional difference in the C-termini of these two proteins.


Assuntos
Proteínas de Bactérias/metabolismo , Biofilmes/crescimento & desenvolvimento , Neoplasias/microbiologia , Streptococcus mutans/patogenicidade , Fatores de Virulência/metabolismo , Animais , Modelos Animais de Doenças , Ratos , Streptococcus mutans/fisiologia
19.
Artigo em Inglês | MEDLINE | ID: mdl-22495251

RESUMO

In this paper, we report on many phosphate containing natural minerals found in the Jenolan Caves - Australia. Such minerals are formed by the reaction of bat guano and clays from the caves. Among these cave minerals is the montgomeryite mineral [Ca(4)MgAl(4)(PO(4))(6)·(OH)(4)·12H(2)O]. The presence of montgomeryite in deposits of the Jenolan Caves - Australia has been identified by X-ray diffraction (XRD). Raman spectroscopy complimented with infrared spectroscopy has been used to characterise the crystal structure of montgomeryite. The Raman spectrum of a standard montgomeryite mineral is identical to that of the Jenolan Caves sample. Bands are assigned to H(2)PO(4)(-), OH and NH stretching vibrations. By using a combination of XRD and Raman spectroscopy, the existence of montgomeryite in the Jenolan Caves - Australia has been proven. A mechanism for the formation of montgomeryite is proposed.


Assuntos
Alumínio/química , Cálcio/química , Cavernas/química , Magnésio/química , Metais/química , Minerais/química , Austrália , Padrões de Referência , Espectrofotometria Infravermelho , Análise Espectral Raman , Difração de Raios X
20.
Artigo em Inglês | MEDLINE | ID: mdl-22446787

RESUMO

The two minerals borickyite and delvauxite CaFe(4)(3+)(PO(4),SO(4))(2)(OH)(8)·4-6H(2)O have the same formula. Are the minerals identical or different? The minerals borickyite and delvauxite have been characterised by Raman spectroscopy. The minerals are related to the minerals diadochite and destinezite. Both minerals are amorphous. Delvauxite appears to vary in crystallinity from amorphous to semi-crystalline. The minerals are often X-ray non-diffracting. The minerals are found in soils and may be described as 'colloidal' minerals. Vibrational spectroscopy enables an assessment of the molecular structure of borickyite and delvauxite. Bands are assigned to phosphate and sulphate stretching and bending modes. Multiple water bending and stretching modes imply that non-equivalent water molecules in the structure exist with different hydrogen bond strengths. The two minerals show differing spectra and must be considered as different minerals.


Assuntos
Minerais/química , Alumínio/química , Cálcio/química , Ferro/química , Magnésio/química , Fosfatos/química , Análise Espectral Raman , Sulfatos/química , Água/química
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