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2.
J Dent Res ; 100(10): 1169-1177, 2021 09.
Article in English | MEDLINE | ID: mdl-34253090

ABSTRACT

Titanium implants present 2 major drawbacks-namely, the long time needed for osseointegration and the lack of inherent antimicrobial properties. Surface modifications and coatings to improve biomaterials can lose their integrity and biological potential when exposed to stressful microenvironments. Graphene nanocoating (GN) can be deposited onto actual-size dental and orthopedic implants. It has antiadhesive properties and can enhance bone formation in vivo. However, its ability to maintain structural integrity and quality when challenged by biologically relevant stresses remains largely unknown. GN was produced by chemical vapor deposition and transferred to titanium via a polymer-assisted transfer technique. GN has high inertness and did not increase expression of inflammatory markers by macrophages, even in the presence of lipopolysaccharides. It kept high coverage at the top tercile of tapered dental implant collars after installation and removal from bone substitute and pig maxilla. It also resisted microbiologically influenced corrosion, and it maintained very high coverage area and quality after prolonged exposure to biofilms and their removal by different techniques. Our findings show that GN is unresponsive to harsh and inflammatory environments and that it maintains a promising level of structural integrity on the top tercile of dental implant collars, which is the area highly affected by biofilms during the onset of implant diseases. Our findings open the avenues for the clinical studies required for the use of GN in the development of implants that have higher osteogenic potential and are less prone to implant diseases.


Subject(s)
Dental Implants , Graphite , Animals , Coated Materials, Biocompatible , Osseointegration , Surface Properties , Swine , Titanium
3.
Benef Microbes ; 12(3): 215-230, 2021 Jun 15.
Article in English | MEDLINE | ID: mdl-34057054

ABSTRACT

Probiotics are thought to be beneficial microbes that influence health-related outcomes through host immunomodulation and modulation of the bacteriome. Its reported success in the treatment of gastrointestinal disorders has led to further research on its potential applicability within the dental field due to similarities such as a polymicrobial aetiology and disease associated microbial-shifts. Although the literature is replete with studies demonstrating its efficacy, the use of probiotics in dentistry continues to polarise opinion. Here, we explore the evidence for probiotics and its effect on periodontal and peri-implant health. MEDLINE, EMBASE, and CENTRAL were systemically searched from June 2010 to June 2020 based on a formulated search strategy. Of 1,956 potentially relevant articles, we selected 27 double-blinded randomised clinical trials in the areas of gingivitis, periodontitis, residual pockets during supportive periodontal therapy, and peri-implant diseases, and reviewed their efficacy in these clinical situations. We observed substantial variation in treatment results and protocols between studies. Overall, the evidence for probiotic therapy for periodontal and peri-implant health appears unconvincing. The scarcity of trials with adequate power and follow-up precludes any meaningful clinical recommendations. Thus, the routine use of probiotics for these purposes are currently unsubstantiated. Further multi-centre trials encompassing a standardised investigation on the most promising strains and administration methods, with longer observation times are required to confirm the benefits of probiotic therapy for these applications.


Subject(s)
Gingivitis/therapy , Peri-Implantitis/therapy , Periodontitis/therapy , Probiotics/therapeutic use , Humans , Immunomodulation , Probiotics/pharmacology , Randomized Controlled Trials as Topic , Stomatitis/therapy , Treatment Outcome
4.
JDR Clin Trans Res ; 6(3): 343-351, 2021 07.
Article in English | MEDLINE | ID: mdl-32777190

ABSTRACT

INTRODUCTION: Previous studies have largely explored the microbial composition and pathogenesis of pregnancy gingivitis. However, the patterns of microbial colonization during pregnancy in the absence of pregnancy gingivitis have rarely been studied. Characterization of the oral microbiome in pregnant women with healthy gingiva is an important initial step in understanding the role of the microbiome in progression to pregnancy gingivitis. OBJECTIVES: In this study, we compared the oral microbiome of pregnant women without gingivitis (healthy pregnancy) with pregnant women having gingivitis and nonpregnant healthy women to understand how pregnancy modifies the oral microbiome and induces progression to pregnancy gingivitis. METHODS: Subgingival plaque samples were collected from Chinese pregnant women with gingivitis (n = 10), healthy pregnant women (n = 10), and nonpregnant healthy women (n = 10). The Illumina MiSeq platform was used to perform 16S rRNA gene sequencing targeting the V4 region. RESULTS: The alpha and beta diversity was significantly different between pregnant and nonpregnant women, but minimal differences were observed between pregnant women with and without gingivitis. Interestingly, the oral bacterial community showed higher abundance of pathogenic taxa during healthy pregnancy as compared with nonpregnant women despite similar gingival and plaque index scores. However, when compared with overt pregnancy gingivitis, pathogenic taxa were less abundant during healthy pregnancy. PICRUSt analysis (phylogenetic investigation of communities by reconstruction of unobserved states) also suggested no difference in the functional capabilities of the microbiome during pregnancy, irrespective of gingival disease status. However, metabolic pathways related to amino acid metabolism were significantly increased in healthy pregnant women as compared with nonpregnant women. CONCLUSION: The presence of pathogenic taxa in healthy pregnancy and pregnancy gingivitis suggests that bacteria may be necessary for initiating disease development but progression to gingivitis may be influenced by the host environmental factors. More efforts are required to plan interventions aimed at sustaining health before the appearance of overt gingivitis. KNOWLEDGE TRANSFER STATEMENT: The results of this study draw attention to the importance of oral health maintenance during pregnancy, as women without any prenatal oral conditions are predisposed to the risk of developing pregnancy gingivitis. Hence, it is important to incorporate comprehensive assessment of oral health in the prenatal health care schedules. Pregnant woman should be screened for oral risks, counseled on proper oral hygiene and expected oral changes, and referred for dental treatment, when necessary.


Subject(s)
Dental Plaque , Gingivitis , Microbiota , Female , Humans , Phylogeny , Pregnancy , RNA, Ribosomal, 16S/genetics
5.
Infect Immun ; 87(10)2019 10.
Article in English | MEDLINE | ID: mdl-31383746

ABSTRACT

Candida albicans, a major opportunistic fungal pathogen, is frequently found together with Streptococcus mutans in dental biofilms associated with severe childhood caries (tooth decay), a prevalent pediatric oral disease. However, the impact of this cross-kingdom relationship on C. albicans remains largely uncharacterized. Here, we employed a novel quantitative proteomics approach in conjunction with transcriptomic profiling to unravel molecular pathways of C. albicans when cocultured with S. mutans in mixed biofilms. RNA sequencing and iTRAQ (isobaric tags for relative and absolute quantitation)-based quantitative proteomics revealed that C. albicans genes and proteins associated with carbohydrate metabolism were significantly enhanced, including sugar transport, aerobic respiration, pyruvate breakdown, and the glyoxylate cycle. Other C. albicans genes and proteins directly and indirectly related to cell morphogenesis and cell wall components such as mannan and glucan were also upregulated, indicating enhanced fungal activity in mixed-species biofilm. Further analyses revealed that S. mutans-derived exoenzyme glucosyltransferase B (GtfB), which binds to the fungal cell surface to promote coadhesion, can break down sucrose into glucose and fructose that can be readily metabolized by C. albicans, enhancing growth and acid production. Altogether, we identified key pathways used by C. albicans in the mixed biofilm, indicating an active fungal role in the sugar metabolism and environmental acidification (key virulence traits associated with caries onset) when interacting with S. mutans, and a new cross-feeding mechanism mediated by GtfB that enhances C. albicans carbohydrate utilization. In addition, we demonstrate that comprehensive transcriptomics and quantitative proteomics can be powerful tools to study microbial contributions which remain underexplored in cross-kingdom biofilms.


Subject(s)
Biofilms/growth & development , Candida albicans/genetics , Gene Expression Regulation, Bacterial , Gene Expression Regulation, Fungal , Streptococcus mutans/genetics , Transcriptome , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Candida albicans/metabolism , Candida albicans/pathogenicity , Carbohydrate Metabolism , Cell Wall/metabolism , Child , Coculture Techniques , Dental Caries/microbiology , Dental Caries/pathology , Glucans/metabolism , Glucosyltransferases/genetics , Glucosyltransferases/metabolism , Humans , Proteomics , Streptococcus mutans/metabolism , Streptococcus mutans/pathogenicity , Symbiosis/genetics
6.
Oral Dis ; 24(4): 518-526, 2018 May.
Article in English | MEDLINE | ID: mdl-28513096

ABSTRACT

Pathobiology of dental caries is complex. Data from recent molecular microbiologic studies have further redefined the role of the oral microbiome in the etiology of dental caries. This new information challenges the conventional view on the hegemony of classic cariogenic prokaryotes such as Streptococcus mutans in caries etiology, and raises the intriguing possibility of the participation of the eukaryotic oral fungal pathogen Candida in the caries process. The virulence attributes of Candida species such as their acidogenicity and aciduric nature, the ability to develop profuse biofilms, ferment and assimilate dietary sugars, and produce collagenolytic proteinases are all indicative of their latent cariogenic potential. Based on the above, oral candidal counts have been used by some as a caries risk indicator. On the contrary, other studies suggest that Candida is merely a passenger extant in an acidic cariogenic milieu, and not a true pathogen. In this review, we critically examine the varying roles of Candida, and traditionally accepted cariogens such as the mutans group of streptococci in the pathobiology of dental caries. The weight of available data tends to imply that Candida may play a pivotal role as a secondary agent perpetuating the carious process, especially in dentinal caries.


Subject(s)
Biofilms , Candida albicans/metabolism , Carbohydrate Metabolism , Dental Caries/microbiology , Streptococcus mutans/metabolism , Acids/metabolism , Candida albicans/enzymology , Humans
7.
J Dent Res ; 96(10): 1129-1135, 2017 Sep.
Article in English | MEDLINE | ID: mdl-28605597

ABSTRACT

Streptococcus mutans is a biofilm-forming oral pathogen commonly associated with dental caries. Clinical studies have shown that S. mutans is often detected with Candida albicans in early childhood caries. Although the C. albicans presence has been shown to enhance bacterial accumulation in biofilms, the influence of S. mutans on fungal biology in this mixed-species relationship remains largely uncharacterized. Therefore, we aimed to investigate how the presence of S. mutans influences C. albicans biofilm development and coexistence. Using a newly established haploid biofilm model of C. albicans, we found that S. mutans augmented haploid C. albicans accumulation in mixed-species biofilms. Similarly, diploid C. albicans also showed enhanced biofilm formation in the presence of S. mutans. Surprisingly, the presence of S. mutans restored the biofilm-forming ability of C. albicans bcr1Δ mutant and bcr1Δ/Δ mutant, which is known to be severely defective in biofilm formation when grown as single species. Moreover, C. albicans hyphal growth factor HWP1 as well as ALS1 and ALS3, which are also involved in fungal biofilm formation, were upregulated in the presence of S. mutans. Subsequently, we found that S. mutans-derived glucosyltransferase B (GtfB) itself can promote C. albicans biofilm development. Interestingly, GtfB was able to increase the expression of HWP1, ALS1, and ALS3 genes in the C. albicans diploid wild-type SC5314 and bcr1Δ/Δ, leading to enhanced fungal biofilms. Hence, the present study demonstrates that a bacterial exoenzyme (GtfB) augments the C. albicans counterpart in mixed-species biofilms through a BCR1-independent mechanism. This novel finding may explain the mutualistic role of S. mutans and C. albicans in cariogenic biofilms.


Subject(s)
Biofilms/growth & development , Candida albicans/physiology , Glucosyltransferases/physiology , Streptococcus mutans/enzymology , Candida albicans/genetics , Coculture Techniques , Colony Count, Microbial , Dental Caries/microbiology , Dental Plaque/microbiology , Gene Expression , Microscopy, Confocal , Reverse Transcriptase Polymerase Chain Reaction , Symbiosis
9.
Int J Nurs Stud ; 51(11): 1472-81, 2014 Nov.
Article in English | MEDLINE | ID: mdl-24647054

ABSTRACT

BACKGROUND: South Asians experience higher rates of cardiovascular disease than any other ethnic group. Some evidence suggests that South Asians may be less adherent to cardiac medication regimens than Whites residing in Canada. Identification of the key factors contributing to adherence may assist this growing population to optimize their cardiac health. AIM: To explore key factors associated with adherence to cardiac medications among South Asian people with cardiac disease. METHODS: Ethnography was used to highlight population specific themes and domains related to medication adherence. Ethnographic observations were undertaken of patients, as well as their family (primary care) physicians and pharmacists (including their staff), while in physician offices and pharmacies. A purposive sample of patients (n=8), as well as physicians (n=3) and pharmacists (n=2) underwent in-depth interviews. Field note and interview data were transcribed verbatim and analyzed using ethnographic domain and thematic analysis. RESULTS: The patients relied on family members for instrumental support in remaining adherent with their medications. Relationships with healthcare professionals who demonstrated clear communication and cultural awareness was associated with enhanced medication adherence. Memory mechanisms needed to be individualized and were generally 'low technology'. While prepackaging of medications enhanced adherence, patients who used them were less knowledgeable about their medications. CONCLUSIONS: Healthcare providers should understand the importance of including family members in the care of South Asian people with heart disease. They also need to appreciate that the quality of provider-patient relationships are important to South Asian patients and will influence adherence to healthcare regimens. Expanding the role of nurse in the primary healthcare team could provide a cost-effective means of enhancing medication adherence.


Subject(s)
Heart Diseases/drug therapy , Patient Compliance , Aged , Alberta , Asia/ethnology , Communication Barriers , Female , Humans , Male , Middle Aged
10.
Clin Oral Investig ; 18(1): 1-16, 2014 Jan.
Article in English | MEDLINE | ID: mdl-23860901

ABSTRACT

OBJECTIVES: To evaluate the effectiveness of natural compounds containing mouthrinse (NCCM) as an adjunct to unsupervised oral hygiene in the management of dental plaque and gingivitis. MATERIALS AND METHODS: An electronic search for clinical studies of NCCMs was conducted in Medline-PubMed, the Cochrane Central Register of Controlled Trials and EMBASE for a period spanning from the earliest available date in each database up to February 2013. Plaque index, gingival index, and gingival bleeding index were selected as primary outcomes. The methodological quality of the studies was assessed according to the "Levels of Evidence" outlined by the Center of Evidence-Based Medicine, and to the Jadad scale. RESULTS: The screen yielded 2,236 titles and abstracts that met the inclusion criteria. These identified 11 clinical trials testing 13 different NCCMs, and were used for data extraction. Heterogeneity and the limited number of studies on any individual NCCM precluded a formal meta-analysis. Of the 13 NCCMs tested, eight demonstrated positive results, and few reported any adverse effects or events. CONCLUSIONS: Evidence proving the effectiveness of NCCM as an adjunct to unsupervised oral hygiene for plaque and gingivitis control is still insufficient. However, some natural products (compounds) may have oral health benefits, so further high-quality study is warranted. CLINICAL RELEVANCE: This review provides an overview of the strength of clinical evidence regarding the effectiveness of natural compounds containing mouthrinses in promoting gingival health.


Subject(s)
Biological Products/therapeutic use , Dental Plaque/drug therapy , Gingivitis/drug therapy , Mouthwashes/therapeutic use , Biological Products/adverse effects , Dental Plaque/prevention & control , Gingivitis/prevention & control , Humans , Mouthwashes/adverse effects , Periodontal Index
11.
Arch Oral Biol ; 58(10): 1327-34, 2013 Oct.
Article in English | MEDLINE | ID: mdl-23880095

ABSTRACT

OBJECTIVE: Enterococcus faecalis is a bacterial pathogen that is often associated with endodontic infections. Biofilm formation is a key virulence attribute in the pathogenicity of E. faecalis. In the present study, we comprehensively examined the effect of various culture media and nutrients on the development of E. faecalis biofilms. DESIGN: A reference strain and a clinical isolate of E. faecalis were used in all experiments for comparison. Commonly used liquid culture media with different nutrient compositions were used to support the development of E. faecalis biofilms in a time-dependent assay. E. faecalis biofilms were quantified by colony forming unit (CFU) and crystal violet (CV) assays. Biofilm architecture and cellular viability were evaluated by scanning electron microscopy and confocal laser scanning microscopy. RESULTS: Growth kinetics evaluated by CFU and CV assays and by microscopy showed that E. faecalis biofilms reached maturity at 72h. "Pg broth" (Tryptic Soy Broth with yeast extract, hemen and vitamin K) promoted E. faecalis biofilm formation more than Brain Heart Infusion broth or Tryptic Soy Broth. Addition of 2% glucose enhanced biofilm formation. Thus, it seems that nutrients such as hemen, vitamin K and glucose are important for E. faecalis for the formation of biofilms. CONCLUSION: The present study demonstrated that nutrient-rich media containing glucose enhances the formation of E. faecalis biofilms, which exhibit maturation at 72h.


Subject(s)
Biofilms/drug effects , Biofilms/growth & development , Culture Media/pharmacology , Enterococcus faecalis/drug effects , Enterococcus faecalis/growth & development , Enterococcus faecalis/pathogenicity , Enterococcus faecalis/ultrastructure , Humans , Microscopy, Confocal , Microscopy, Electron, Scanning , Virulence
12.
Chin Med J (Engl) ; 126(4): 634-9, 2013 Feb.
Article in English | MEDLINE | ID: mdl-23422180

ABSTRACT

BACKGROUND: Few literatures pertain to the 16S ribosomal DNA (16S rDNA) analysis of bacteria contributing to primary and persistent endodontic lesions, with no information available for the Chinese population. As such, we investigated endodontic bacteria associated with primary and persistent endodontic lesions in adult Chinese patients living in Beijing, China using 16S rDNA gene sequencing techniques. METHODS: Endodontic microbial samples were obtained from fourteen adult Chinese patients and subjected to DNA extraction. Pllymerase chain reaction (PCR) products were cloned and 100 clones from each generated library were randomly selected. Purified plasmid DNA with 16S rDNA gene inserts was sequenced, and the sequences were searched against GenBank databases using the BLASTN algorithm. Only significant identification with the highest-scored BLAST result and 99% minimum similarity was considered for phylotyping. RESULTS: More than 150 taxa were obtained. Primary endodontic infection was mainly associated with Burkholderia cepacia, Actinomyces, Aranicola spp. and Streptococcus sanguinis, whilst Burkholderia cepacia was predominant in the persistent endodontic infections. CONCLUSION: There is a difference in the species profile associated with endodontic infections of Chinese patients living in Beijing in comparison to other geographical or ethnic reports.


Subject(s)
Bacteria/pathogenicity , DNA, Ribosomal/genetics , Pulpitis/microbiology , Sequence Analysis, DNA/methods , Bacteria/classification , Bacteria/genetics , China , DNA, Bacterial/genetics , Female , Humans , Male
13.
J Hosp Infect ; 81(2): 79-86, 2012 Jun.
Article in English | MEDLINE | ID: mdl-22595316

ABSTRACT

BACKGROUND: Most clinical guidelines for the use of biocides have been developed for planktonic micro-organisms, but in nature, most micro-organisms live as surface-adherent communities or biofilms. AIM: To evaluate the effectiveness of commonly used biocides against Escherichia coli and Candida spp. in three distinct growth phases: planktonic, adhesion and biofilm. METHODS: Ultrastructural, architectural and cellular viability changes following a 5 min exposure to biocide were monitored by scanning electron microscopy and confocal laser scanning microscopy using fluorescent dyes. Comparative transcript expression of the antioxidants SOD1 and CAT1 in the planktonic and biofilm phases was evaluated using quantitative real-time polymerase chain reaction. FINDINGS: E. coli and Candida spp. in the planktonic phase were susceptible to all the tested biocides at the recommended concentrations. However, early adhesion and late biofilm phases of both were less susceptible to the biocides, and exceeded the recommended concentrations on several occasions. A short period of biocide exposure failed to fully eradicate the adherent microbial cells, and they recovered from the biocide challenge, forming biofilm on the biocide-treated surfaces. The biofilm phase showed higher expression of SOD1 and CAT1. CONCLUSION: The recommended concentrations of biocides for clinical disinfection in the hospital setting may not fully eradicate the adhesion or biofilm phases of E. coli and Candida spp. Higher antioxidative capacities in microbial biofilms may be responsible for the resistance of biofilms against clinical biocides.


Subject(s)
Biofilms/drug effects , Candida/physiology , Disinfectants/pharmacology , Drug Resistance, Bacterial , Drug Resistance, Fungal , Escherichia coli/physiology , Candida/drug effects , Candida/ultrastructure , Catalase/biosynthesis , Escherichia coli/drug effects , Escherichia coli/ultrastructure , Gene Expression Profiling , Humans , Microbial Sensitivity Tests , Microbial Viability/drug effects , Microscopy, Confocal , Microscopy, Electron, Scanning , Oxidative Stress , Stress, Physiological , Superoxide Dismutase/biosynthesis , Superoxide Dismutase-1
14.
Proteomics ; 12(4-5): 651-65, 2012 Feb.
Article in English | MEDLINE | ID: mdl-22246638

ABSTRACT

Biofilms are surface-attached, matrix-encased, structured microbial communities which display phenotypic features that are dramatically different from those of their free-floating, or planktonic, counterparts. Biofilms seem to be the preferred mode of growth of microorganisms in nature, and at least 65% of all human infections are associated with biofilms. The most notable and clinically relevant property of biofilms is their greater resistance to antimicrobials compared with their planktonic counterparts. Although both bacterial and fungal biofilms display this phenotypic feature, the exact mechanisms underlying their increased drug resistance are yet to be determined. Advances in proteomics techniques during the past decade have facilitated in-depth analysis of the possible mechanisms underpinning increased drug resistance in biofilms. These studies have demonstrated the ability of proteomics techniques to unravel new targets for combating microbial biofilms. In this review, we discuss the putative drug resistance mechanisms of microbial biofilms that have been uncovered by proteomics and critically evaluate the possible contribution of the new knowledge to future development in the field. We also summarize strategic uses of novel proteomics technologies in studies related to drug resistance mechanisms of microbial biofilms.


Subject(s)
Bacteria/drug effects , Biofilms/drug effects , Drug Resistance, Fungal , Drug Resistance, Multiple, Bacterial , Fungi/drug effects , Proteomics/methods , Bacterial Infections/drug therapy , Bacterial Infections/metabolism , Bacterial Infections/microbiology , Bacterial Physiological Phenomena , Biofilms/growth & development , Fungi/physiology , Humans , Oxidative Stress , Quorum Sensing
15.
Am J Orthod Dentofacial Orthop ; 140(4): 537-42, 2011 Oct.
Article in English | MEDLINE | ID: mdl-21967942

ABSTRACT

INTRODUCTION: The aim of this study was to evaluate the antimicrobial efficacy of 2 commercially available mouth rinses on a monospecies-biofilm model on orthodontic brackets in vitro. METHODS: The antimicrobial effects of the 2 mouth rinses, Listerine (tartar control; IDS Manufacturing, Bangkok, Thailand) and Corsodyl (SmithKline Beecham, Maidenhead, United Kingdom), on the planktonic Streptococcus mutans were tested by maximum inhibitory dilution assay. The cell viability of S mutans biofilm on Damon3 MX brackets (Ormco, Glendora, Calif) after exposure to the 2 mouth rinses was quantified by 2,3-bis (2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino) carbonyl]-2H-tetrazolium hydroxide (XTT) reduction assay. Visualization of the biofilm samples was performed by fluorescence microscopy and confocal laser scanning microscopy. RESULTS: The maximum inhibitory dilution assays of S mutans were 1:5 for Listerine and 1:320 for Corsodyl. The optical density values, which were measured by XTT reduction assay from S mutans biofilms after 1 minute of exposure to the different test agents, demonstrated that the cell viability of S mutans biofilms exposed to Listerine was less than that for Corsodyl, which was less than that for brain-heart infusion (P <0.001). Listerine caused more dead cells on the surface of the brackets than did Corsodyl when examined with the 2 microscope systems. CONCLUSIONS: Both mouth rinses showed marked antimicrobial effects on the monospecies biofilm in vitro. Listerine showed a stronger bactericidal effect but had less bacterial inhibitory effect than did Corsodyl.


Subject(s)
Anti-Infective Agents, Local/pharmacology , Biofilms/drug effects , Chlorhexidine/analogs & derivatives , Mouthwashes/pharmacology , Orthodontic Brackets/microbiology , Salicylates/pharmacology , Streptococcus mutans/drug effects , Terpenes/pharmacology , Anti-Infective Agents, Local/administration & dosage , Bacterial Load , Bacteriological Techniques , Chlorhexidine/administration & dosage , Chlorhexidine/pharmacology , Drug Combinations , Humans , Indicators and Reagents , Materials Testing , Microbial Viability/drug effects , Microscopy, Confocal , Microscopy, Fluorescence , Mouthwashes/administration & dosage , Salicylates/administration & dosage , Streptococcus mutans/physiology , Terpenes/administration & dosage , Tetrazolium Salts , Time Factors
16.
J Anal Toxicol ; 35(7): 431-7, 2011 Sep.
Article in English | MEDLINE | ID: mdl-21871151

ABSTRACT

Methadone is used worldwide for the treatment of heroin addiction; however, fatal poisonings are increasingly reported. The prevalence of CYP2B6 and µ-opioid receptor (OPRM1) gene variations were examined between a postmortem population where the deaths were associated with methadone and a live nondrug-using control population using Taqman™ SNP Genotyping assays. The CYP2B6*6 allele was higher in the postmortem population, but the difference was not significant (P = 0.92). The CYP2B6 T750C promoter variation was similar in frequency for both populations. Linkage between T750C and CYP2B6*6 was identified for both populations (P < 0.01). The prevalence of the OPRM1 A118G variation was significantly higher in the control population (P = 0.0046), which might indicate a protective mechanism against opioid toxicity. Individual susceptibility to methadone may be determined by screening for CYP2B6*6.


Subject(s)
Aryl Hydrocarbon Hydroxylases/genetics , Forensic Genetics , Genetic Predisposition to Disease , Methadone/poisoning , Oxidoreductases, N-Demethylating/genetics , Polymorphism, Single Nucleotide , Receptors, Opioid, mu/genetics , Adolescent , Adult , Age Factors , Case-Control Studies , Cytochrome P-450 CYP2B6 , Female , Gene Frequency , Genetic Testing , Genotype , Humans , Male , Methadone/pharmacokinetics , Middle Aged , Poisoning/genetics , Poisoning/mortality , Prevalence , Sex Factors , Young Adult
17.
Mycopathologia ; 172(5): 389-95, 2011 Nov.
Article in English | MEDLINE | ID: mdl-21744043

ABSTRACT

Candida bloodstream infection has dramatically increased in the last decade due to the growing number of immunocompromised populations worldwide. In this study, we evaluated the antifungal susceptibility profiles and virulence attributes of Candida bloodstream isolates (CBIs) derived from Hong Kong and Finland, information which are vital for devising empirical clinical strategies. Susceptibility testing of a wide range of antifungals including fluconazole, itraconazole, voriconazole, ketoconazole, 5-fluorocytosine, amphotericin B and caspofungin was performed. Haemolytic activity and secretion of proteinase of CBIs were also examined. All CBIs derived from Hong Kong were susceptible to all the antifungals tested whilst some CBIs from Finland were resistant to azoles and caspofungin. C. albicans, C. glabrata and C. tropicalis showed higher haemolytic activity whereas C. parapsilosis and C. guilliermondii were non-haemolytic in general. Proteinase activity of the Finland C. albicans isolates was significantly higher than the Hong Kong isolates. Our data provide a glimpse of the possible evolutionary changes in pathogenic potential of Candida that may be occurring in different regions of the world. Therefore, continuous surveillance and availability of local data should be taken into consideration when treating candidemia patients.


Subject(s)
Antifungal Agents/pharmacology , Candida/drug effects , Candida/pathogenicity , Candidemia/microbiology , Virulence Factors/metabolism , Amphotericin B/pharmacology , Candida/isolation & purification , Caspofungin , Drug Resistance, Fungal , Echinocandins/pharmacology , Finland , Fluconazole/pharmacology , Flucytosine/pharmacology , Hemolysin Proteins/metabolism , Hong Kong , Humans , Itraconazole/pharmacology , Ketoconazole/pharmacology , Lipopeptides , Microbial Sensitivity Tests , Peptide Hydrolases/metabolism , Pyrimidines/pharmacology , Triazoles/pharmacology , Voriconazole
18.
Electrophoresis ; 32(12): 1492-9, 2011 Jun.
Article in English | MEDLINE | ID: mdl-21563180

ABSTRACT

In this paper, we demonstrate, using both experiment and simulation, how sample zone conductivity can affect plug-plug mixing in small molecule applications of electrophoretically mediated microanalysis (EMMA). The effectiveness of in-line mixing, which is driven by potential, can vary widely with experimental conditions. Using two small molecule systems, the effects of local conductivity differences between analyte plugs, reagent plugs and the BGE on EMMA analyses are examined. Simul 5.0, a dynamic simulation program for CE systems, is used to understand the ionic boundaries and profiles that give rise to the experimentally obtained data for EMMA analyses for (i) creatinine determination via the Jaffe reaction, a reaction involving a neutral and an anion, and (ii) the redox reaction between gallate and 2,6-dichloroindophenol, two anions. Low sample conductivity, which is widely used in CE analyses, can be detrimental for in-line reactions involving a neutral reactant, as rapid migration of the ionic component across a low conductivity neutral zone results in poor reagent plug overlap and low reaction efficiency. Conversely, with two similarly charged reagents, a low conductivity sample plug is advantageous, as it allows field-amplified stacking of the reagents into a tight reaction zone. In addition, the complexity of simultaneously overlapping three reagent zones is considered, and experimental results validate the predictions made by the simulation. The simulations, however, do not appear to predict all of the observed experimental behavior. Overall, by combining experiment with simulation, an enhanced appreciation for the local field effects in EMMA is realized, and general guidelines for an advantageous sample matrix can be established for categories of EMMA analyses.


Subject(s)
Electric Conductivity , Electrophoresis, Capillary/methods , Models, Chemical , 2,6-Dichloroindophenol/chemistry , Anions/chemistry , Computer Simulation , Creatinine/chemistry , Gallic Acid/chemistry , Picrates/chemistry
19.
J Med Microbiol ; 60(Pt 9): 1241-1247, 2011 Sep.
Article in English | MEDLINE | ID: mdl-21474609

ABSTRACT

Biofilm formation is a major virulence attribute of Candida albicans and is directly associated with therapeutic failure. One method by which Candida acquires antifungal resistance is the expression of drug-resistance genes. This study aimed to evaluate the transcriptional regulation of several genes associated with antifungal resistance of C. albicans under planktonic, recently adhered and biofilm growth modes and in C. albicans biofilms in response to antifungal agents. Initially, the antifungal susceptibility of C. albicans cultures in different growth modes was evaluated by standard antifungal susceptibility testing. Next, to assess CDR1, CDR2, MDR1, ERG11, FKS1 and PIL1 expression, RNA was harvested from cells in each growth mode, and from biofilms after drug treatment, and subjected to quantitative real-time RT-PCR (qRT-PCR). Biofilm C. albicans was more resistant to antifungals than recently adhered cells and stationary-phase planktonic cultures. Transcriptional expression of CDR1, CDR2, MDR1, ERG11 and FKS1 was lower in recently adhered C. albicans than in the stationary-phase planktonic cultures. In contrast, PIL1 levels were significantly increased in recently adhered and biofilm modes of growth. The expression of MDR1 in biofilms greatly increased on challenge with amphotericin B but not with the other drugs tested (P<0.01). ERG11 was significantly upregulated by ketoconazole (P<0.01). Caspofungin and amphotericin B significantly upregulated FKS1 expression, whereas they significantly downregulated PIL1 expression (P<0.01). These results indicate that the expression of drug-resistance genes is associated with higher drug resistance of Candida biofilms, and lay a foundation for future large-scale genome-wide expression analysis.


Subject(s)
Antifungal Agents/pharmacology , Biofilms/drug effects , Candida albicans/drug effects , Drug Resistance, Fungal , Gene Expression Regulation, Fungal/drug effects , Transcription, Genetic , Transcriptional Activation , Antifungal Agents/metabolism , Biofilms/growth & development , Candida albicans/physiology , Fungal Proteins/biosynthesis , Fungal Proteins/genetics , Gene Expression Profiling , Microbial Sensitivity Tests
20.
Arch Oral Biol ; 56(1): 16-21, 2011 Jan.
Article in English | MEDLINE | ID: mdl-20864088

ABSTRACT

OBJECTIVE: the aim of this study was to evaluate the antimicrobial efficacy of Traditional Chinese Medicine Fructus mume on a monospecies-biofilm model established on orthodontic brackets in vitro. MATERIALS AND METHODS: the antimicrobial effect of Fructus mume aqueous extract on the planktonic Streptococcus mutans (S. mutans) was tested by microdilution method (MIC). The cell viability of S. mutans biofilm on Damon3 MX bracket (Ormco, USA) after exposed to Fructus mume extract was quantified by XTT reduction assay. Visualization of the samples was performed by fluorescence microscope and confocal laser scanning microscopy (CLSM). RESULTS: HPLC analysis revealed that the main compounds of Fructus mume are organic acids. The MIC of Fructus mume extract on the planktonic S. mutans was 50mg/mL. The optical density (OD) values, measured by XTT reduction assay from S. mutans biofilms after 1-min exposure to different test agents, demonstrated that the cell viability of S. mutans biofilms exposed to 250mg/mL Fructus mume extract

Subject(s)
Anti-Bacterial Agents/pharmacology , Biofilms/drug effects , Drugs, Chinese Herbal/pharmacology , Orthodontic Brackets/microbiology , Plant Extracts/pharmacology , Prunus , Chromatography, High Pressure Liquid , Citric Acid/analysis , Drugs, Chinese Herbal/chemistry , Humans , Indicators and Reagents , Materials Testing , Microbial Sensitivity Tests , Microbial Viability/drug effects , Microscopy, Confocal , Microscopy, Fluorescence , Oxalic Acid/analysis , Plant Extracts/analysis , Prunus/chemistry , Spectrum Analysis , Streptococcus mutans/drug effects , Tartrates/analysis , Tetrazolium Salts
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