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1.
Animal ; 15(2): 100121, 2021 Feb.
Article in English | MEDLINE | ID: mdl-33712212

ABSTRACT

The work presented in this pilot study aimed to identify potential risk factors associated with bovine periodontitis development. Bovine periodontitis is a multifactorial polymicrobial infectious disease for which the aetiopathogenesis and risk factors are not fully understood. From cattle slaughtered in an abattoir in Scotland, 35 dental arcades with periodontal lesions and 40 periodontally healthy arcades were selected over seven visits for study. Multivariable logistic regression analysis was used to evaluate the association between periodontitis and the independent variables, gender, age and breed. For every increase in year of age, cattle were 1.5 times more likely to have periodontitis. A graphical analysis indicated that within the limits of this study, we could not detect any major influence of breed on the age-effect. Although logistic regression analysis demonstrated that periodontitis lesions are more prevalent with increasing age of cattle the underlying mechanisms remain unclear. It is likely that periodontitis is an important cause of oral pain in older cattle and can contribute to reduced productivity/performance. Further studies with a larger sample size are necessary to elucidate the associations between potential risk factors and periodontitis in cattle and to define its effects on animal welfare and productivity.


Subject(s)
Cattle Diseases , Periodontitis , Animals , Cattle , Cattle Diseases/epidemiology , Periodontitis/epidemiology , Periodontitis/veterinary , Pilot Projects , Risk Factors , Scotland/epidemiology
3.
Mar Environ Res ; 155: 104881, 2020 Mar.
Article in English | MEDLINE | ID: mdl-32072985

ABSTRACT

Given the severity of injuries to biota in coastal wetlands from the Deepwater Horizon oil spill (DWH) and the resulting availability of funding for restoration, information on impacted salt marshes and biotic development of restored marshes may both help inform marsh restoration planning in the near term and for future spills. Accordingly, we performed a meta-analysis to model a restoration trajectory of total macroinfauna density in constructed marshes (studied for ~30 y), and with a previously published restoration trajectory for amphipods, we compared these to recovery curves for total macroinfauna and amphipods from DWH impacted marshes (over 8.5 y). Total macroinfauna and amphipod densities in constructed marshes did not consistently reach equivalency with reference sites before 20 y, yet in heavily oiled marshes recovery occurred by 4.5 y post spill (although it is unlikely that macroinfaunal community composition fully recovered). These differences were probably due to initial conditions (e.g., higher initial levels of belowground organic matter in oiled marshes) that were more conducive to recovery as compared to constructed marshes. Furthermore, we found that amphipod trajectories were distinctly different in constructed and oiled marshes as densities at oiled sites exceeded that of reference sites by as much as 20x during much of the recovery period. Amphipods may have responded to the rapid increase and high biomass of benthic microalgae following the spill. These results indicate that biotic responses after an oil spill may be quantitatively different than those following restoration, even for heavily oiled marshes that were initially denuded of vegetation. Our dual trajectories for oil spill recovery and restoration development for macroinfauna should help guide restoration planning and assessment following the DWH as well as for restoration scaling for future spills.


Subject(s)
Amphipoda/growth & development , Environmental Restoration and Remediation , Petroleum Pollution , Water Pollution, Chemical , Wetlands , Animals , Biomass , Gulf of Mexico , Models, Biological
4.
Proc Natl Acad Sci U S A ; 116(38): 18917-18922, 2019 09 17.
Article in English | MEDLINE | ID: mdl-31484762

ABSTRACT

Photosystem II (PSII) performs the solar-driven oxidation of water used to fuel oxygenic photosynthesis. The active site of water oxidation is the oxygen-evolving complex (OEC), a Mn4CaO5 cluster. PSII requires degradation of key subunits and reassembly of the OEC as frequently as every 20 to 40 min. The metals for the OEC are assembled within the PSII protein environment via a series of binding events and photochemically induced oxidation events, but the full mechanism is unknown. A role of proton release in this mechanism is suggested here by the observation that the yield of in vitro OEC photoassembly is higher in deuterated water, D2O, compared with H2O when chloride is limiting. In kinetic studies, OEC photoassembly shows a significant lag phase in H2O at limiting chloride concentrations with an apparent H/D solvent isotope effect of 0.14 ± 0.05. The growth phase of OEC photoassembly shows an H/D solvent isotope effect of 1.5 ± 0.2. We analyzed the protonation states of the OEC protein environment using classical Multiconformer Continuum Electrostatics. Combining experiments and simulations leads to a model in which protons are lost from amino acid that will serve as OEC ligands as metals are bound. Chloride and D2O increase the proton affinities of key amino acid residues. These residues tune the binding affinity of Mn2+/3+ and facilitate the deprotonation of water to form a proposed µ-hydroxo bridged Mn2+Mn3+ intermediate.


Subject(s)
Chlorides/chemistry , Oxygen/metabolism , Photosystem II Protein Complex/metabolism , Water/chemistry , Catalytic Domain , Deuterium , Kinetics , Manganese/chemistry , Oxidation-Reduction , Photosystem II Protein Complex/chemistry , Protons , Solvents/chemistry , Solvents/metabolism , Static Electricity , Water/metabolism
5.
Equine Vet J ; 49(3): 294-299, 2017 May.
Article in English | MEDLINE | ID: mdl-27270960

ABSTRACT

REASONS FOR PERFORMING STUDY: Equine periodontitis is a common and painful condition. However, the disease often goes unnoticed by owners and is thus a major welfare concern. The aetiopathogenesis of the condition remains poorly understood and has been investigated in few studies. The innate immune system is known to play an important role in human periodontitis, but its role in equine periodontitis has not been examined. OBJECTIVES: To quantify the messenger (m)RNA levels of Toll-like receptors (TLRs) and cytokines in gingival tissue from orally healthy horses and those affected by periodontitis. STUDY DESIGN: Observational study. METHODS: Gingival tissue samples were taken post-mortem from 13 horses with no clinical signs of oral disease and 20 horses with periodontitis. mRNA levels of TLR2, TLR4 and TLR9 and cytokines interleukin-1ß (IL-1ß), tumour necrosis factor-α (TNF-α), IL-4, IL-6, IL-10, IL-12, IL-17 and interferon-γ (IFN-γ) were determined using quantitative real-time PCR. The statistical significance of results was assessed using appropriate t tests. RESULTS: mRNA levels of all TLRs and cytokines were upregulated in equine periodontitis. Significant increases in mRNA levels of TLR2, TLR9, IL-4, IL-10, IL-12 (P≤0.05) and IFN-γ (P≤0.01) were observed for both unweighted and age-weighted analyses of diseased gingival tissue samples compared with healthy gingival samples. In comparisons of samples of periodontitis lesions with healthy gingival control samples from the same horse, significant increases in mRNA levels of TLR4, TLR9, IL-10, IFN-γ (P≤0.05), TLR2, IL-1ß and IL-12p35 (P≤0.01) were observed. CONCLUSIONS: This study has provided an initial insight into the involvement of the immune system in equine periodontitis. Increased mRNA levels of TLR2, TLR4 and TLR9 indicate substantial microbial challenge in diseased gingival tissue. A mixed Th1/Th2/Th17 cytokine response is produced in equine periodontitis. Further studies are required to more fully characterise the role of the innate immune system in this disease.


Subject(s)
Cytokines/metabolism , Horse Diseases/metabolism , Periodontitis/veterinary , RNA, Messenger/metabolism , Toll-Like Receptors/metabolism , Animals , Cytokines/genetics , Female , Gene Expression Regulation , Horses , Male , Periodontitis/metabolism , Periodontitis/pathology , RNA, Messenger/genetics , Toll-Like Receptors/genetics
7.
Clin Oral Investig ; 20(9): 2529-2537, 2016 Dec.
Article in English | MEDLINE | ID: mdl-26888221

ABSTRACT

OBJECTIVES: The serum IL-17A:IL-17E ratio has previously been demonstrated to be a clinical marker of periodontitis. The aim of this study was to determine the effects of non-surgical periodontal treatment on the serum IL-17A:IL-17E ratio. MATERIALS AND METHODS: Forty chronic periodontitis patients completed this study and received periodontal treatment comprising scaling and root planing plus ultrasonic debridement. Clinical data were recorded at baseline, 6 weeks (R1) after treatment completion (full-mouth or quadrant-scaling and root planing) and 25 weeks after baseline (R2). Serum samples were taken at each time point and cytokines concentrations determined by ELISA. RESULTS: Following treatment, statistically significant reductions were noted in clinical parameters. However, IL-17A and IL-17E concentrations were significantly greater than baseline values before- and after-adjusting for smoking. The IL-17A:IL-17E ratio was lower at R1 and R2. Serum IL-6 and TNF levels were significantly lower at R1 only. Also exclusively at R1, serum IL-17A and IL-17E correlated positively with clinical parameters, while the IL-17A:IL-17E ratio correlated negatively with probing pocket depth and clinical attachment. CONCLUSION: Increased serum IL-17E and a reduced IL-17A:IL-17E ratio may be indicative and/or a consequence of periodontal therapy. Therefore, the role of IL-17E in periodontal disease progression and the healing process is worthy of further investigation. CLINICAL RELEVANCE: IL-17E may be a valuable biomarker to monitor the healing process following periodontal treatment as increased IL-17E levels and a reduced IL-17A:IL-17E ratio could reflect clinical improvements post-therapy. Therefore, monitoring serum IL-17E might be useful to identify individuals who require additional periodontal treatment.


Subject(s)
Chronic Periodontitis/therapy , Dental Scaling , Interleukin-17/blood , Root Planing , Adult , Biomarkers/blood , Debridement , Enzyme-Linked Immunosorbent Assay , Female , Humans , Male , Periodontal Index
8.
Int J Sports Med ; 36(1): 49-53, 2015 Jan.
Article in English | MEDLINE | ID: mdl-25254901

ABSTRACT

We compared the accuracy of 2 GPS systems with different sampling rates for the determination of distances covered at high-speed and metabolic power derived from a combination of running speed and acceleration. 8 participants performed 56 bouts of shuttle intermittent running wearing 2 portable GPS devices (SPI-Pro, GPS-5 Hz and MinimaxX, GPS-10 Hz). The GPS systems were compared with a radar system as a criterion measure. The variables investigated were: total distance (TD), high-speed distance (HSR>4.17 m·s(-1)), very high-speed distance (VHSR>5.56 m·s(-1)), mean power (Pmean), high metabolic power (HMP>20 W·kg(-1)) and very high metabolic power (VHMP>25 W·kg(-1)). GPS-5 Hz had low error for TD (2.8%) and Pmean (4.5%), while the errors for the other variables ranged from moderate to high (7.5-23.2%). GPS-10 Hz demonstrated a low error for TD (1.9%), HSR (4.7%), Pmean (2.4%) and HMP (4.5%), whereas the errors for VHSR (10.5%) and VHMP (6.2%) were moderate. In general, GPS accuracy increased with a higher sampling rate, but decreased with increasing speed of movement. Both systems could be used for calculating TD and Pmean, but they cannot be used interchangeably. Only GPS-10 Hz demonstrated a sufficient level of accuracy for quantifying distance covered at higher speeds or time spent at very high power.


Subject(s)
Geographic Information Systems , Running/physiology , Soccer/physiology , Time and Motion Studies , Acceleration , Adolescent , Energy Metabolism , Humans , Male , Reproducibility of Results
9.
Scand J Med Sci Sports ; 22(3): 392-8, 2012 Jun.
Article in English | MEDLINE | ID: mdl-20807389

ABSTRACT

This study analyzes the anthropometric and physiological characteristics of junior cyclists within different cycling specialties and different performance levels. One hundred and thirty-two junior riders (16.8 ± 0.6 years, 177 ± 6 cm, 66.3 ± 6.7 kg) were tested for anthropometric, aerobic and anaerobic parameters. Cyclists were classified within specialties [uphill (UH) flat terrain (FT) all terrain (AT) and sprint (SP)] and performance levels, based on a seasonal ranking [low level (LL) medium level (ML) and high level (HL)]. The results of the two-way analysis of variance showed that FT and SP have greater body dimensions than UH and AT (P<0.001). Concerning the relative aerobic parameters, AT and UH have higher values (P<0.001) than FT and SP [maximal oxygen uptake (VO(2max) ): 69.4 ± 3.6, 67.5 ± 5.0, 62.8 ± 4.5 and 61.9 ± 4.1 mL/kg/min, respectively] while absolute parameters resulted higher for FT and AT (P≤0.008). The relative power produced in the 5 s test was higher (P<0.001) for AT and SP than FT and UH (16.7 ± 1.1, 16.6 ± 0.6, 14.9 ± 1.7 and 14.4 ± 1.7 W/kg, respectively). Concerning the performance level, only the age and the aerobic parameters resulted differently within levels (VO(2max) : HL=67.3 ± 4.9, ML=65.5 ± 5.1 and LL=63.3 ± 5.2 mL/kg/min), with the highest values for HL (P≤0.007). In conclusion, juniors are specialized in the same way as professional cyclists and the aerobic characteristics are confirmed as significant in the performance level assessment.


Subject(s)
Anthropometry , Bicycling/physiology , Adolescent , Analysis of Variance , Athletic Performance/physiology , Exercise Test , Female , Humans , Male , Oxygen Consumption/physiology , Physical Endurance/physiology , Physical Fitness/physiology , Statistics, Nonparametric
10.
Eur J Clin Microbiol Infect Dis ; 29(7): 823-34, 2010 Jul.
Article in English | MEDLINE | ID: mdl-20449620

ABSTRACT

The purpose of this study was to use microbiological culture and bacterial 16S rRNA gene sequencing methods to detect transcriptionally active bacteria present on the surface of failed prosthetic hip joints removed during revision arthroplasty. Five failed prosthetic hip joints were sonicated to dislodge adherent bacteria and subjected to microbiological culture. Bacterial RNA was extracted from each sonicate, cDNA prepared by reverse transcription and the 16S rRNA gene amplified using universal primers. Polymerase chain reaction (PCR) products were cloned, assigned to distinct groups by restriction fragment length polymorphism (RFLP) analysis and one representative clone from each group was sequenced. Bacteria were identified by comparison of the obtained 16S rRNA gene sequences with those deposited in public access sequence databases. All five specimens were positive for the presence of bacteria by both culture and PCR. Culture methods identified species from eight genera. Molecular detection of transcriptionally active bacteria identified a wider range of species. A total of 42 phylotypes were identified, of which Lysobacter gummosus was the most abundant (31.6%). Thirty-four clones (14.5%) represented uncultivable phylotypes. No potentially novel species were identified. It is concluded that a diverse range of transcriptionally active bacterial species are present within biofilms on the surface of failed prosthetic hip joints.


Subject(s)
Arthroplasty/adverse effects , Bacteria/isolation & purification , Hip Joint/surgery , Microbial Viability , Prosthesis-Related Infections/microbiology , Transcription, Genetic , Adult , Aged , Bacteria/classification , Bacteria/genetics , Bacteria/growth & development , Cloning, Molecular , DNA Fingerprinting , DNA, Bacterial/genetics , DNA, Bacterial/metabolism , DNA, Complementary/genetics , DNA, Complementary/metabolism , Humans , Male , Middle Aged , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , RNA, Bacterial/genetics , RNA, Bacterial/isolation & purification , RNA, Ribosomal, 16S/genetics , RNA, Ribosomal, 16S/isolation & purification , Reverse Transcriptase Polymerase Chain Reaction , Sequence Analysis, DNA
11.
Environ Entomol ; 38(4): 1174-81, 2009 Aug.
Article in English | MEDLINE | ID: mdl-19689897

ABSTRACT

Rice, Oryza sativa, is the most important staple food for a significant portion of the world's population. Despite the importance of rice, however, induced resistance to insects has not been thoroughly studied in rice; in fact, to our knowledge, direct induced resistance after injury by chewing insects has not been shown in rice. We conducted a series of experiments designed to characterize direct induced resistance in rice after feeding by larvae of the fall armyworm (Spodoptera frugiperda J. E. Smith) and application of jasmonic acid. Weight gains and relative growth rates of fall armyworm larvae were lower when fed leaves from plants previously damaged by armyworms than when fed leaves from undamaged plants. This response was stronger at a systemic spatial scale; that is, the induced resistance was stronger in newly emerged leaves not present at the time plants were damaged than in damaged leaves themselves. Armyworm growth rates were also reduced on foliage from plants treated with jasmonic acid, a hormone known to mediate plant responses to wounding. The response to injury by armyworm larvae and to exogenous jasmonic acid was stronger in transgenic rice plants in which levels of salicylic acid (a signaling molecule that inhibits jasmonic acid) were suppressed. These results show the existence of a direct induced resistance response in rice and suggest that this response to injury by a chewing insect may be mediated by jasmonic acid.


Subject(s)
Cyclopentanes/metabolism , Host-Parasite Interactions , Oryza/immunology , Oxylipins/metabolism , Salicylic Acid/metabolism , Spodoptera/physiology , Animals , Larva/physiology , Oryza/genetics , Oryza/metabolism , Plants, Genetically Modified/immunology , Plants, Genetically Modified/metabolism
12.
Eur J Clin Invest ; 38(10): 713-20, 2008 Oct.
Article in English | MEDLINE | ID: mdl-18837796

ABSTRACT

BACKGROUND: Atherosclerosis is potentiated by stimulation of Toll-like receptors (TLRs), which serve to detect pathogen associated molecular patterns (PAMPs). However little is known of which PAMPs may be present in atheroma, or capable of stimulating inflammatory signalling in vascular cells. MATERIALS AND METHODS: DNA extracted from human carotid atheroma samples was amplified and sequenced using broad-range 16S gene specific primers to establish historical exposure to bacterial PAMPs. Responsiveness of primary human arterial and venous endothelial and smooth muscle cells to PAMPs specific for each of the TLRs was assessed by measurement of interleukin-8 secretion and E-selectin expression. RESULTS: Extracts of atheromatous tissue stimulated little or no signalling in TLR-transfected HEK-293 cells. However, sequencing of bacterial DNA amplified from carotid atheroma revealed the presence of DNA from 17 different bacterial genera, suggesting historical exposure to bacterial lipopeptide, lipopolysaccharide and flagellin. All cells examined were responsive to the ligands of TLR3 and TLR4, poly inosine:cytosine and lipopolysaccharide. Arterial cells were responsive to a wider range of PAMPs than venous cells, being additionally responsive to bacterial flagellin and unmethylated cytosine-phosphate-guanosine DNA motifs, the ligands of TLR5 and TLR9, respectively. Cells were generally unresponsive towards the ligands of human TLR7 and TLR8, loxoribine and single stranded RNA. Only coronary artery endothelial cells expressed TLR2 mRNA and responded to the TLR2 ligand Pam(3)CSK(4). CONCLUSIONS: Vascular cells are responsive to a relatively diverse range of TLR ligands and may be exposed, at least transiently, to ligands of TLR2, TLR4, TLR5 and TLR9 during the development of carotid atheroma.


Subject(s)
Antigens, Bacterial/pharmacology , Carotid Artery Diseases/metabolism , Endothelial Cells/immunology , Toll-Like Receptors/metabolism , Antigens, Bacterial/immunology , Biomarkers/analysis , Carotid Artery Diseases/immunology , Cell Line , DNA Primers/genetics , DNA, Bacterial/analysis , E-Selectin/analysis , Endothelial Cells/drug effects , Humans , Interleukin-8/analysis , Ligands , Myocytes, Smooth Muscle/immunology , Reverse Transcriptase Polymerase Chain Reaction , Sequence Analysis, DNA , Toll-Like Receptor 2/genetics , Toll-Like Receptor 2/metabolism , Toll-Like Receptor 4/genetics , Toll-Like Receptor 4/metabolism , Toll-Like Receptor 5/genetics , Toll-Like Receptor 5/metabolism , Toll-Like Receptor 9/genetics , Toll-Like Receptor 9/metabolism , Toll-Like Receptors/genetics , Toll-Like Receptors/immunology , Transfection/methods
13.
Oral Dis ; 14(3): 251-8, 2008 Apr.
Article in English | MEDLINE | ID: mdl-18336372

ABSTRACT

AIM: Compare the microbial profiles on the tongue dorsum in patients with halitosis and control subjects in a UK population using culture-independent techniques. MATERIALS AND METHODS: Halitosis patients were screened according to our recently developed recruitment protocol. Scrapings from the tongue dorsum were obtained for 12 control subjects and 20 halitosis patients. Bacteria were identified by PCR amplification, cloning and sequencing of 16S rRNA genes. RESULTS: The predominant species found in the control samples were Lysobacter-type species, Streptococcus salivarius, Veillonella dispar, unidentified oral bacterium, Actinomyces odontolyticus, Atopobium parvulum and Veillonella atypica. In the halitosis samples, Lysobacter-type species, S. salivarius, Prevotella melaninogenica, unidentified oral bacterium, Prevotella veroralis and Prevotella pallens were the most commonly found species. For the control samples, 13-16 (4.7-5.8%) of 276 clones represented uncultured species, whereas in the halitosis samples, this proportion increased to 6.5-9.6% (36-53 of 553 clones). In the control samples, 22 (8.0%) of 276 clones represented potentially novel phylotypes, and in the halitosis samples, this figure was 39 (7.1%) of 553 clones. CONCLUSIONS: The microflora associated with the tongue dorsum is complex in both the control and halitosis groups, but several key species predominate in both groups.


Subject(s)
Halitosis/microbiology , Tongue/microbiology , Bacterial Typing Techniques , Biofilms , Case-Control Studies , DNA, Bacterial/analysis , Gram-Positive Asporogenous Rods/isolation & purification , Humans , Liver Transplantation , Polymerase Chain Reaction , Prevotella/isolation & purification , RNA, Ribosomal, 16S/analysis , Sequence Analysis, DNA
14.
Oral Dis ; 13(1): 63-70, 2007 Jan.
Article in English | MEDLINE | ID: mdl-17241432

ABSTRACT

OBJECTIVE: To develop and apply a detailed clinical protocol for screening and assessing subjects with a complaint of halitosis. DESIGN: Cross-sectional. SUBJECTS AND METHODS: Several methods were used to recruit subjects with a complaint of halitosis, including a newspaper advertisement. A definition of halitosis arising from within the oral cavity, which is not related to generalized chronic gingivitis, chronic periodontitis or pathology of the oral mucosa was used. An extensive list of exclusion criteria was applied at the initial visit. Eligible subjects were asked to follow strict instructions and complete a questionnaire prior to their second visit for data collection. The clinical examination consisted of an organoleptic assessment, Halimeter reading and periodontal examination. RESULTS: The best method of recruiting subjects was advertising. Of 66 individuals recruited, four failed to attend the screening visit and 25 were excluded. The main reasons for exclusion were poor oral hygiene and existing periodontal disease. Thirty-seven completed the full protocol, resulting in identification of 18 with halitosis and 19 controls. CONCLUSIONS: Application of the exclusion criteria resulted in significant attrition of eligible participants. Our results suggest that organoleptic assessment should be regarded as a useful standard for defining subjects with halitosis.


Subject(s)
Halitosis/diagnosis , Adolescent , Adult , Cross-Sectional Studies , Female , Humans , Lung , Male , Mass Screening , Middle Aged , Mouth , Nose , Odorants/analysis , Oral Hygiene , Patient Selection , Periodontal Diseases/diagnosis , Smell , Surveys and Questionnaires
15.
J Clin Periodontol ; 32(9): 973-83, 2005 Sep.
Article in English | MEDLINE | ID: mdl-16104962

ABSTRACT

OBJECTIVES: The aim of the current study was to assess the impact of smoking on the clinical indices, the humoral immune response and the detection frequency of putative periodontal pathogens in patients with periodontitis cross-sectionally and following therapy. MATERIAL AND METHODS: Clinical measurements, subgingival plaque samples, gingival crevicular fluid (GCF) and sera were collected from 40 untreated patients with moderate-to-advanced chronic periodontitis before and after treatment over a period of 6 months. The treatment consisted of the initial therapy of scaling and root planing. Smoking status was self-reported and was confirmed by cotinine enzyme inhibition assay (CEIA). Whole-mouth clinical measurements were recorded with a manual periodontal probe at baseline (BAS) and at 6 months (RAS). Selected-site analyses were performed on the deepest site in each quadrant before and after therapy and clinical indices were recorded with an electronic pressure-sensitive probe. GCF sample volume was quantified using the Periotron 6000. Polymerase chain reaction (PCR) was utilized to determine the presence of Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, Prevotella intermedia, Treponema denticola and Tanerella forsythensis in subgingival plaque. Enzyme-linked immunosorbent assay examined the systemic antibody titres to these bacteria, and thiocyanate disassociation determined the antibody avidity to these organisms. RESULTS: At baseline, smokers showed significantly less gingival inflammation and lower GCF volume compared with non-smokers. After treatment, a compromised clinical outcome was noted for smokers in terms of pocket depth reduction and gain in attachment levels. No significant differences in the detection of putative periodontal pathogens in subgingival plaque existed between smokers and non-smokers. A consistent trend was noted in that smokers had lower sera immunoglobulin G antibody titres to these organisms before and after treatment (statistically significant for A. actinomycetemcomitans). This pattern was less clear when antibody avidities were considered, revealing only small differences, if any, between the two groups of patients. CONCLUSION: Current data indicate that smokers with periodontal disease have a suppressed inflammatory response, a significantly less favourable clinical outcome and seem to have an altered host antibody response to antigenic challenge than non-smokers. In contrast, the subgingival microflora of smokers appears similar to that of non-smokers.


Subject(s)
Periodontitis , Smoking/adverse effects , Adult , Aged , Chi-Square Distribution , Cross-Sectional Studies , Female , Gingival Crevicular Fluid/microbiology , Humans , Male , Middle Aged , Periodontitis/blood , Periodontitis/immunology , Periodontitis/microbiology , Polymerase Chain Reaction , Statistics, Nonparametric
16.
Oral Dis ; 11 Suppl 1: 61-3, 2005.
Article in English | MEDLINE | ID: mdl-15752102

ABSTRACT

OBJECTIVE: Determination of the microflora present on the tongue dorsum of subjects with and without halitosis using conventional microbiological culture methods. METHODS: Twenty-one halitosis and 20 control patients were recruited using a strict clinical protocol. Samples were collected from the posterior dorsum of the tongue using a sterile brush. Each sample was vortex mixed for 30 s and serial 10-fold dilutions to 10(-7) were carried out. Samples were plated onto fastidious anaerobe agar (FAA) and FAA enriched with vancomycin. These were incubated under anaerobic conditions for 10 days at 37 degrees C. Strict anaerobes were identified by metronidazole sensitivity and bacteria were identified to genus level by a combination of colony morphology, Gram staining and biochemical and enzymatic tests (rapid ID 32 A). RESULTS: The predominant species in test and control groups were Veillonella sp. and Prevotella sp. Greater species diversity was found in the halitosis samples compared with controls. The halitosis samples contained an increased incidence of unidentifiable Gram-negative rods, Gram-positive rods and Gram-negative coccobacilli. CONCLUSIONS: There was no obvious association between halitosis and any specific bacterial genus. The increased species diversity found in halitosis samples suggests that halitosis may be the result of complex interactions between several bacterial species. The role of uncultivable bacteria may also be important in contributing to this process.


Subject(s)
Halitosis/microbiology , Tongue/microbiology , Bacteria, Anaerobic/isolation & purification , Bacterial Typing Techniques , Case-Control Studies , Colony Count, Microbial , Humans
17.
J Dent ; 33(4): 293-303, 2005 Apr.
Article in English | MEDLINE | ID: mdl-15781137

ABSTRACT

UNLABELLED: Some dental procedures initiate a bacteraemia. In certain compromised patients, this bacteraemia may lead to distant site infections, most notably infective endocarditis. OBJECTIVE: To investigate whether a detectable bacteraemia was produced during non-surgical root canal therapy. METHODS: Thirty patients receiving non-surgical root canal therapy were studied. Three blood samples were taken per patient: pre-operatively, peri-operatively and post-operatively. In addition, a paper point sample was collected from the root canal. The blood samples were cultured by pour plate and blood bottle methods. The isolated organisms were identified by standard techniques. Blood samples were analysed for the presence of bacterial DNA by the polymerase chain reaction (PCR). In two cases where the same species of organism was identified in the root canal and the bloodstream, the isolates were typed by pulsed field gel electrophoresis (PFGE). RESULTS: By conventional culturing, a detectable bacteraemia was present in 9 (30%) of the 30 patients who had no positive pre-operative control blood sample. In 7 (23.3%) patients, the same species of organism was identified in both the bloodstream and in the paper point sample from the root canal system. Overall, PCR gave lower detection rates compared with conventional culture, with 10 of 90 (11%) of the blood samples displaying bacterial DNA. PFGE typing was undertaken for two pairs of culture isolates from blood and paper points; these were found to be genetically identical. CONCLUSIONS: Non-surgical root canal treatment may invoke a detectable bacteraemia.


Subject(s)
Bacteremia/diagnosis , Root Canal Therapy , Anti-Infective Agents, Local/therapeutic use , Bacteremia/microbiology , Bacteria/classification , Bacteriological Techniques , Chlorhexidine/therapeutic use , Dental Cavity Preparation/instrumentation , Dental Cavity Preparation/methods , Dental Pulp Cavity/microbiology , Electrophoresis, Agar Gel , Electrophoresis, Gel, Pulsed-Field , Female , Gutta-Percha/therapeutic use , Humans , Hydrogen Peroxide/therapeutic use , Male , Middle Aged , Polymerase Chain Reaction , Potassium Iodide/therapeutic use , Root Canal Filling Materials/therapeutic use , Root Canal Irrigants/therapeutic use , Root Canal Obturation , Root Canal Therapy/methods , Rubber Dams , Sodium Hypochlorite/therapeutic use
18.
J Clin Periodontol ; 32(2): 200-6, 2005 Feb.
Article in English | MEDLINE | ID: mdl-15691352

ABSTRACT

OBJECTIVES: To compare the effects of scaling and root planing (SRP) on clinical and microbiological parameters at selected sites in smoker and non-smoker chronic and generalized aggressive periodontitis patients. MATERIALS AND METHODS: Clinical parameters including probing depth (PD), relative attachment level (RAL), and bleeding upon probing (BOP), and subgingival plaque samples were taken from four sites in 28 chronic periodontitis (CP) and 17 generalized aggressive periodontitis (GAgP) patients before and after SRP. Polymerase chain reaction assays were used to determine the presence of A. actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythensis, Prevotella intermedia and Treponema denticola. RESULTS: Both CP and GAgP non-smokers had significantly greater reduction in pocket depth (1.0+/-1.3 mm in CP smokers versus 1.7+/-1.4 mm in non-smokers, p=0.007 and 1.3+/-1.0 in GAgP smokers versus 2.4+/-1.2 mm in GAgP non-smokers, p<0.001) than respective non-smokers, with a significant decrease in Tannerella forsythensis in CP sites (smokers 25% increase and non-smokers 36.3% decrease, p<0.001) and Prevotella intermedia at GAgP sites (smokers 25% reduction versus 46.9% in non-smokers, p=0.028). CONCLUSION: SRP was effective in reducing clinical parameters in both groups. The inferior improvement in PD following therapy for smokers may reflect the systemic effects of smoking on the host response and the healing process. The lesser reduction in microflora and greater post-therapy prevalence of organisms may reflect the deeper pockets seen in smokers and poorer clearance of the organisms. These detrimental consequences for smokers appear consistent in both aggressive and CP.


Subject(s)
Dental Plaque/microbiology , Periodontitis/microbiology , Smoking/adverse effects , Adult , Bacteroides/genetics , Chi-Square Distribution , Chronic Disease , Dental Plaque/genetics , Dental Plaque/therapy , Dental Scaling/methods , Female , Humans , Male , Middle Aged , Periodontitis/genetics , Periodontitis/therapy , Polymerase Chain Reaction/methods , Root Planing/methods , Statistics, Nonparametric
19.
J Med Microbiol ; 54(Pt 3): 299-303, 2005 Mar.
Article in English | MEDLINE | ID: mdl-15713615

ABSTRACT

Lactobacillus fermentum is a Gram-positive bacterium that is associated with active caries lesions. Methods for identifying Lactobacillus species traditionally have been based upon culture methods coupled with biochemical tests, which are generally unreliable. The aim of this study was to develop a species-specific PCR assay for the direct detection of L. fermentum in oral clinical samples. PCR primers specific for L. fermentum were identified by alignment of bacterial 16S rRNA genes and selection of sequences specific for L. fermentum at their 3' ends. PCR positivity for L. fermentum DNA was indicated by amplification of a 337 bp product. The primers were shown to be specific for L. fermentum DNA, since no PCR product was obtained when genomic DNA from a wide range of other oral bacteria, including closely related Lactobacillus species, were used as test species. The PCR assay was then used in an attempt to identify L. fermentum DNA in supragingival plaque samples and in pus aspirates from subjects with acute dento-alveolar abscesses. Four out of 70 (5.7 %) supragingival plaque samples analysed were positive for the presence of L. fermentum DNA while none of the 19 pus samples analysed was positive for L. fermentum DNA. This PCR assay provides a more rapid, specific and sensitive alternative to conventional culture methods for the identification of L. fermentum in clinical specimens.


Subject(s)
DNA, Bacterial/analysis , Lactobacillus/isolation & purification , Polymerase Chain Reaction/methods , DNA, Ribosomal/analysis , Dental Plaque/microbiology , Humans , Lactobacillus/genetics , Periapical Abscess/microbiology , Polymerase Chain Reaction/standards , Polymorphism, Restriction Fragment Length , RNA, Ribosomal, 16S/genetics , Sensitivity and Specificity , Species Specificity
20.
J Clin Periodontol ; 31(2): 141-8, 2004 Feb.
Article in English | MEDLINE | ID: mdl-15016040

ABSTRACT

OBJECTIVES: The aim of this study was to test the hypothesis that over a period of 6 months, same-day full-mouth scaling and root planing (FM-SRP) resulted in greater reductions in the detection frequency of five putative periodontal pathogens compared with quadrant scaling and root planing (Q-SRP) in chronic periodontitis patients. MATERIALS AND METHODS: Forty patients were recruited into this study. Subjects were randomised into two groups. The FM-SRP group received full-mouth scaling and root planing completed within the same day, while the Q-SRP group received quadrant root planing at 2-weekly intervals over four consecutive sessions. Selected-site analyses were performed on the deepest site in each quadrant before and after therapy, at approximately 3 and 6 months from baseline (R1 and R2) and clinical indices were recorded with an electronic pressure-sensitive probe. In addition, subgingival plaque samples were collected from these sites at baseline (BAS), at reassessment 1 (R1), approximately 6 weeks after the completion of therapy and at reassessment 2 (R2), 6 months from baseline. Polymerase chain reaction (PCR) was used to determine the presence of Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, Prevotella intermedia, Treponema denticola and Bacteroides forsythus in plaque. RESULTS: Both therapies resulted in significant improvements in all clinical indices both at R1 and R2. A marked reduction in the presence of all candidate periodontal pathogens was noted after both treatment modalities, reaching statistical significance for the majority of the test organisms. These improvements were maintained over a period of 6 months. When the two treatment groups were compared, a significantly higher percentage of Q-SRP patients was positive for P. intermedia at R1 compared with FM-SRP patients (p<0.05). In addition, a greater reduction in the patient prevalence for T. denticola was found for the FM-SRP group than the Q-SRP group at R1 and R2 from baseline (p<0.005), but the significance of this is questionable given the skewed detection frequency of this organism at baseline between the two treatments (p<0.01). CONCLUSION: This study failed to confirm that same-day FM-SRP resulted in greater microbiological improvements compared with Q-SRP at 2-weekly intervals over a 6-month period, as determined by PCR.


Subject(s)
Dental Scaling/methods , Periodontitis/microbiology , Root Planing/methods , Adult , Aged , Aggregatibacter actinomycetemcomitans/isolation & purification , Bacteroides/isolation & purification , Chi-Square Distribution , Chronic Disease , Colony Count, Microbial , Dental Plaque/microbiology , Follow-Up Studies , Humans , Middle Aged , Periodontal Attachment Loss/microbiology , Periodontal Attachment Loss/therapy , Periodontal Index , Periodontal Pocket/microbiology , Periodontal Pocket/therapy , Periodontitis/therapy , Porphyromonas gingivalis/isolation & purification , Prevotella intermedia/isolation & purification , Treponema/isolation & purification
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