text:
AIM: To look for clinical signs of periodontal disease in young adults who exhibited radiographic bone loss and detectable numbers of Aggregatibacter actinomycetemcomitans in their primary dentition. MATERIAL AND METHODS: Periodontal status and radiographic bone loss were examined in each of the subjects 16 years after the baseline observations. Techniques for anaerobic and selective culture, and checkerboard, were used to detect periodontitis-associated bacterial species. The isolated A. actinomycetemcomitans strains were characterized by polymerase chain reaction. RESULTS: Signs of localized attachment loss were found in three out of the 13 examined subjects. A. actinomycetemcomitans was recovered from six of these subjects and two of these samples were from sites with deepened probing depths and attachment loss. Among the isolated A. actinomycetemcomitans strains, serotypes a-c and e, but not d or f, were found. None of the isolated strains belonged to the highly leucotoxic JP2 clone, and one strain lacked genes for the cytolethal distending toxin. CONCLUSIONS: This study indicates that the presence of A. actinomycetemcomitans and early bone loss in the primary dentition does not necessarily predispose the individual to periodontal attachment loss in the permanent dentition.
In this study, the essential oil of Origanum vulgare was evaluated for putative antibacterial activity against six clinical strains and five reference strains of Aggregatibacter actinomycetemcomitans, in comparison with some antimicrobials. The chemical composition of the essential oil was analyzed, using chromatography (CG) and gas chromatography-mass spectrometry coupled (CG-MS). The major compounds in the oil were Carvacrol (32.36%), α-terpineol (16.70%), p-cymene (16.24%), and Thymol (12.05%). The antimicrobial activity was determined by an agar well diffusion test. A broth microdilution method was used to study the minimal inhibitory concentration (MIC). The minimal bactericidal concentration (MBC) was also determined. The cytotoxicity of the essential oil (IC50) was <125 µg/mL for THP-1 cells, which was high in comparison with different MIC values for the A. actinomycetemcomitans strains. O. vulgare essential oil did not interfere with the neutralizing capacity of Psidium guajava against the A. actinomycetemcomitans leukotoxin. In addition, it was shown that the O. vulgare EO had an antibacterial effect against A. actinomycetemcomitans on a similar level as some tested antimicrobials. In view of these findings, we suggest that O.vulgare EO may be used as an adjuvant for prevention and treatment of periodontal diseases associated to A. actinomycetemcomitans. In addition, it can be used together with the previously tested leukotoxin neutralizing Psidium guajava.
Obesity and associated metabolic disorders have been implicated in liver carcinogenesis; however there is little data on the role of obesity-related biomarkers on liver cancer risk. We studied prospectively the association of inflammatory and metabolic biomarkers with risks of hepatocellular carcinoma (HCC), intra-hepatic bile duct (IBD) and gallbladder and bilary tract cancers outside of the liver (GBTC) in a nested case-control study within the European Prospective Investigation into Cancer and Nutrition (EPIC). Over an average of 7.7 years, 296 participants developed HCC (n=125), GBTC (n=137) or IBD (n=34). Using risk set sampling, controls were selected in a 2:1 ratio and matched for recruitment center, age, sex, fasting status, time of blood collection. Baseline serum concentrations of C-reactive protein (CRP), interleukin-6 (IL-6), C-peptide, total, high-molecular-weight (HMW) adiponectin, leptin, fetuin-a, and glutamatdehydrogenase (GLDH) were measured and incidence rate ratios (IRRs) and 95% confidence intervals (CI-s) estimated using conditional logistic regression. After adjustment for lifestyle factors, diabetes, hepatitis infection and adiposity measures, higher concentrations of CRP, IL-6, C-peptide and non-HMW adiponectin were associated with higher risk of HCC (IRR per doubling of concentrations = 1.22; 95%CI = 1.02-1.46, P=0.03; 1.90; 95%CI = 1.30-2.77, P=0.001; 2.25; 95%CI = 1.43-3.54, P=0.0005 and 2.09; 95%CI = 1.19-3.67, P=0.01, respectively). CRP was associated also with risk of GBTC (IRR = 1.22; 95%CI = 1.05-1.42, P=0.01). GLDH was associated with risks of HCC (IRR = 1.62; 95%CI = 1.25-2.11, P=0.0003) and IBD (IRR = 10.5; 95%CI = 2.20-50.90, P=0.003). The continuous net reclassification index was 0.63 for CRP, IL-6, C-peptide and non-HMW adiponectin, and 0.46 for GLDH indicating good predictive ability of these biomarkers. Conclusion: Elevated levels of biomarkers of inflammation and hyperinsulinemia are associated with a higher risk of HCC, independent of obesity and established liver cancer risk factors.
BACKGROUND: The chewing stick, the miswak, is used in many developing countries as the traditional means for oral hygiene. It is prepared from the roots, twigs and stem of Salvadora persica or other alternative local plants. OBJECTIVES: To compare the effects of the chewing stick miswak (from S. persica) and toothbrush on subgingival plaque microflora among Saudi Arabian individuals. Further, to investigate whether components extracted from S. persica may interfere with the subgingival plaque micro-organisms. MATERIAL AND METHODS: Fifteen healthy Saudi Arabian male volunteers aged 21-36 years were included in a single-blind, randomized cross-over study. The participants were taught how to use each device properly. Plaque sampling for DNA test was performed at the baseline, 1 week after professional tooth cleaning, and after 3 weeks of either miswak or toothbrush use. Identification and quantification of microbial species were performed by the checkerboard method, using whole genomic, digoxigenin-labelled DNA probes. Inhibition zones around miswak were examined on agar plates with Actinobacillus actinomycetemcomitans and the leukotoxicity of this bacterium was analyzed in a bioassay with macrophages+/-extracts of miswak. RESULTS: Miswak and toothbrushing had a similar influence on the levels of the subgingival microbiota. However, A. actinomycetemcomitans was significantly more reduced by miswak (p<0.05) than by toothbrushing. These results were supported by our in vitro results which, indicated that extracts from S. persica might interfere with the growth and leukotoxicity of A. actinomycetemcomitans. CONCLUSIONS: In contrast to toothbrush use, miswak use significantly reduced the amount of A. actinomycetemcomitans in the subgingival plaque.
A major problem with anti-cancer drug treatment is the development of acquired multidrug resistance (MDR) of the tumor cells. Verotoxin-1 (VT-1) exerts its cytotoxicity by targeting the globotriaosylceramide membrane receptor (Gb3), a glycolipid associated with multidrug resistance. Gb3 is overexpressed in many human tumors and tumor cell lines with inherent or acquired MDR. Gb3 is co-expressed and interplays with the membrane efflux transporter P-gp encoded by the MDR1 gene. P-gp could act as a lipid flippase and stimulate Gb3 induction when tumor cells are exposed to cancer chemotherapy. Recent work has shown that apoptosis and inherent or acquired multidrug resistance in Gb3-expressing tumors could be affected by VT-1 holotoxin, a sub-toxic concentration of the holotoxin concomitant with chemotherapy or its Gb3-binding B-subunit coupled to cytotoxic or immunomodulatory drug, as well as chemical manipulation of Gb3 expression. The interplay between Gb3 and P-gp thus gives a possible physiological approach to augment the chemotherapeutic effect in multidrug resistant tumors.
Cytolethal distending toxin, a bacterial exotoxin produced by a number of Gram-negative species, causes growth arrest and morphological alterations in host cells. Among these species are Haemophilus ducreyi, the etiological agent of chancroid, and the periodontal pathogen Aggregatibacter actinomycetemcomitans, highly implicated in localized aggressive periodontitis. CDT induces receptor activator of NF-kappaB ligand (RANKL) expression in periodontal fibroblasts, the key bone-resorbing cytokine. T-cells are actively involved in localized inflammation-induced bone destruction, including periodontitis. The aim of this study was to investigate the effects of purified CDT on the expression of RANKL and its decoy receptor osteoprotegerin (OPG), in the Jurkat T-cell line. Quantitative real-time PCR indicated that 100 pg/ml of purified H. ducreyi CDT upregulated RANKL mRNA expression by 2.2-fold, after 24 h of exposure. This increase was corroborated by a 2.0-fold increase in RANKL protein release, as determined by ELISA. OPG was not detected in this experimental system. In conclusion, CDT enhances RANKL expression in T-cells, denoting that these cells are a potential target for the toxin and strengthening the potential link between this virulence factor and mechanisms associated with localized bone resorption.
Periodontitis is an inflammatory condition that destroys the tooth-supporting tissues, as a result of local bacterial infection. Aggregatibacter actinomycetemcomitans is a Gram-negative facultative anaerobic species, highly associated with aggressive periodontitis. Periodontal inflammation is dominated by cytokines of the Interleukin (IL)-1 family. Prior to their secretion by mononuclear cells, IL-1 cytokines are processed by intracellular protein complexes, known as "inflammasomes", which can sense the bacterial challenge. The aim of this study was to investigate which inflammasomes are regulated in mononuclear cells in response to A. actinomycetemcomitans. The D7SS strain and its derivative leukotoxin and cytolethal distending toxin knock-out mutant strains were used to infect human mononuclear cells at a 1:10 cell: bacteria ratio, for 3h. The expression of various inflammasome components in the cells was investigated by TaqMan quantitative real-time polymerase chain reaction (qPCR). The expressions of NOD-like receptor protein (NLRP)1, NLRP2 and Absent In Melanoma (AIM)2 inflammasome sensors, as well as their effector Caspase-1 were not affected. However, NLRP3 was up-regulated, while NLRP6 was down-regulated. This effect was not dependent on the leukotoxin or the cytolethal distending toxin, as demonstrated by the use of specific gene knock-out mutant strains. IL-1β and IL-18 expressions were also up-regulated by the bacterial challenge. In conclusion, A. actinomycetemcomitans enhances NLRP3 and reduces NLRP6 inflammasome expression, irrespective of its major virulence factors, confirming the high pathogenic profile of this species, and providing further insights to the mechanisms of periodontal inflammation.
Actinobacillus actinomycetemcomitans is associated with localized aggressive periodontitis, a disease characterized by rapid loss of the alveolar bone surrounding the teeth. Receptor activator of NF-kappaB Ligand (RANKL) and osteoprotegerin (OPG) are two molecules that regulate osteoclast formation and bone resorption. RANKL induces osteoclast differentiation and activation, whereas OPG blocks this process by acting as a decoy receptor for RANKL. The purpose of this study was to investigate the effect of A. actinomycetemcomitans on the expression of RANKL and OPG in human gingival fibroblasts and periodontal ligament cells. RANKL mRNA expression was induced in both cell types challenged by A. actinomycetemcomitans extract, whereas OPG mRNA expression remained unaffected. Cell surface RANKL protein was also induced by A. actinomycetemcomitans, whereas there was no change in OPG protein secretion. A cytolethal distending toxin (Cdt) gene-knockout strain of A. actinomycetemcomitans did not induce RANKL expression, in contrast to its wild-type strain. Purified Cdt from Haemophilus ducreyi alone, or in combination with extract from the A. actinomycetemcomitans cdt mutant strain, induced RANKL expression. Pretreatment of A. actinomycetemcomitans wild-type extract with Cdt antiserum abolished RANKL expression. In conclusion, A. actinomycetemcomitans induces RANKL expression in periodontal connective tissue cells. Cdt is crucial for this induction and may therefore be involved in the pathological bone resorption during the process of localized aggressive periodontitis.
Actinobacillus actinomycetemcomitans is implicated in the pathogenesis of localized aggressive periodontitis, and has the capacity to express a cytolethal distending toxin (Cdt). Gingival fibroblasts (GF) are resident cells of the periodontium, which can express several osteolytic cytokines. The aims of this study were a) to investigate the role of Cdt in A. actinomycetemcomitans-induced expression of osteolytic cytokines and their cognate receptors in GF and b) to determine if the previously demonstrated induction of receptor activator of NFkappaB ligand (RANKL) by A. actinomycetemcomitans is mediated by these pro-inflammatory cytokines or by prostaglandin E(2) (PGE(2)). A. actinomycetemcomitans clearly induced interleukin (IL)-6, IL-1beta, and to a minimal extent, tumor necrosis factor (TNF)-alpha mRNA expression. At the protein level, IL-6 but not IL-1beta or TNF-alpha expression was stimulated. The mRNA expression of the different receptor subtypes recognizing IL-6, IL-1beta and TNF-alpha was not affected. A cdt-knockout strain of A. actinomycetemcomitans had similar effects on cytokine and cytokine receptor mRNA expression, compared to its parental wild-type strain. Purified Cdt stimulated IL-6, but not IL-1beta or TNF-alpha protein biosynthesis. Antibodies neutralizing IL-6, IL-1 or TNF-alpha, and the PGE(2) synthesis inhibitor indomethacin, did not affect A. actinomycetemcomitans-induced RANKL expression. In conclusion, a) A. actinomycetemcomitans induces IL-6 production in GF by a mechanism largely independent of its Cdt and b) A. actinomycetemcomitans-induced RANKL expression in GF occurs independently of IL-1, IL-6, TNF-alpha, or PGE(2).
The cytolethal distending toxin (Cdt) is produced by several Gram-negative bacterial species and causes growth arrest and morphological alterations in mammalian cells. Actinobacillus actinomycetemcomitans, which is involved in the pathogenesis of localized aggressive periodontitis, also produces a Cdt that affects periodontal connective tissue cells. The aim of this study was to investigate in which phase of the cell cycle these cells are arrested and enlarged when challenged with A. actinomycetemcomitans, and to evaluate the involvement of its Cdt. Human gingival fibroblasts and periodontal ligament cells were challenged with A. actinomycetemcomitans extract, or with purified Cdt, and cell cycle analysis was performed by propidium iodide staining and flow cytometry. Cells exposed to an A. actinomycetemcomitans wild-type strain, or to purified Cdt, were arrested in both G1 and G2/M phases, and appeared enlarged compared to the corresponding controls. The cellular enlargement occurred in both G1 and G2/M arrested cells. In contrast, cells exposed to an A. actinomycetemcomitans cdt-knockout mutant strain showed cell cycle phase distribution and size similar to the controls. In conclusion, A. actinomycetemcomitans causes a combined G1 and G2/M growth arrest and enlargement in periodontal connective tissue cells, which is attributed to its Cdt.
Aggregatibacter actinomycetemcomitans is a periodontal pathogen colonizing the oral cavity of a large proportion of the human population. It is equipped with several potent virulence factors that can cause cell death and induce or evade inflammation. Because of the large genetic diversity within the species, both harmless and highly virulent genotypes of the bacterium have emerged. The oral condition and age, as well as the geographic origin of the individual, influence the risk to be colonized by a virulent genotype of the bacterium. In the present review, the virulence and pathogenicity properties of A. actinomycetemcomitans will be addressed.
Porphyromonas gingivalis is highly implicated in the pathogenesis of periodontitis, which is characterized by the destruction of periodontal connective tissues and the supporting alveolar bone. Receptor Activator of NF-kappaB Ligand (RANKL) stimulates bone resorption, whereas osteoprotegerin (OPG) blocks its action, and this bi-molecular system is implicated in periodontitis. The aim of this work was (a) to investigate the regulation of RANKL and OPG gene expression in human periodontal ligament (PDL) cells and gingival fibroblasts (GF), in response to P. gingivalis culture supernatants, by quantitative real-time PCR and (b) to attempt to identify putative virulence factors involved in this process. The results indicated that P. gingivalis induced RANKL and reduced OPG mRNA expression by the studied cells, resulting in an increased RANKL/OPG expression ratio. Heat-inactivation of P. gingivalis resulted in significant reduction of RANKL mRNA expression. A Lys-gingipain mutant strain did not affect, whereas an Arg-gingipain mutant strain further enhanced RANKL mRNA expression, compared to their parental wild-type strain. In conclusion, P. gingivalis up-regulates the RANKL/OPG expression ratio in GF and PDL cells, denoting an enhanced osteoclastogenic potential by the cells. The component mainly responsible for RANKL induction appears to be proteinaceous, and it may be regulated by the Arg-gingipains.
BACKGROUND: Coronary artery ectasia (CAE) is a rare disorder commonly associated with additional features of atherosclerosis. In the present study, we aimed to examine the systemic immune-inflammatory response that might associate CAE.
METHODS: Plasma samples were obtained from 16 patients with coronary artery ectasia (mean age 64.9 ± 7.3 years, 6 female), 69 patients with coronary artery disease (CAD) and angiographic evidence for atherosclerosis (age 64.5 ± 8.7 years, 41 female), and 140 controls (mean age 58.6 ± 4.1 years, 40 female) with normal coronary arteries. Samples were analyzed at Umeå University Biochemistry Laboratory, Sweden, using the V-PLEX Pro-Inflammatory Panel 1 (human) Kit. Statistically significant differences (p < 0.05) between patient groups and controls were determined using Mann-Whitney U-tests.
RESULTS: The CAE patients had significantly higher plasma levels of INF-γ, TNF-α, IL-1β, and IL-8 (p = 0.007, 0.01, 0.001, and 0.002, respectively), and lower levels of IL-2 and IL-4 (p < 0.001 for both) compared to CAD patients and controls. The plasma levels of IL-10, IL-12p, and IL-13 were not different between the three groups. None of these markers could differentiate between patients with pure (n = 6) and mixed with minimal atherosclerosis (n = 10) CAE.
CONCLUSIONS: These results indicate an enhanced systemic pro-inflammatory response in CAE. The profile of this response indicates activation of macrophages through a pathway and trigger different from those of atherosclerosis immune inflammatory response.
Porphyromonas gingivalis strongly correlates with periodontitis, but the underlying mechanisms causing dentoalveolar bone resorption are not fully understood. As contradictory effects of P. gingivalis on osteoclastogenesis have been reported, this study investigates the effect of P. gingivalis extract on osteoclast formation. Osteoclast formation in mouse bone marrow (MBM) cell cultures and RAW 264.7 cells was stimulated by nuclear factor-κB ligand (RANKL) or parathyroid hormone (PTH). Cells were cultured with and without P. gingivalis surface-associated material and phenotypic characteristics were examined using microscopy, flow cytometry, and RT-PCR. P. gingivalis significantly decreased osteoclast formation and the expression of osteoclast phenotypic markers in PTH-stimulated MBM cultures. Additionally, P. gingivalis inhibited expression of osteoclast differentiation factors and stimulated expression of the mouse macrophage marker F4/80. The presence of P. gingivalis in RANKL-stimulated MBM cultures and RAW 264.7 cells inhibited osteoclastogenesis. Interestingly, a transient exposure with P. gingivalis before PTH stimulation increased osteoclastogenesis in MBM cultures. Flow cytometric analyses of cells transiently exposed to P. gingivalis demonstrated an increased proportion of potential osteoclast precursor cells. We conclude that a transient exposure of MBM cultures to P. gingivalis increases the number of osteoclast precursors and osteoclast formation, whereas a prolonged exposure completely abolishes osteoclastogenesis.
Background and Objective: Periodontal disease is an inflammatory condition caused by bacterial infections that result in loss of the tooth supporting tissue. The periodontal pathogens produce virulence factors with capacity to affect the host immune response. Aggregatibacter actinomycetemcomitans is a periodontal pathogen that produces a leukotoxin that specifically affects human leukocytes. The aims of the present study were to examine the presence and function of systemic antibodies to the leukotoxin. Material and Methods: One hundred and ninety-seven middle-aged (57 ± 5 years) Swedes with well-documented periodontal status and medical factors related to cardiovascular diseases were studied. These data have been published previously. The serum samples were examined for the presence of leukotoxin antibodies by western blot and the capacity to neutralize leukotoxicity in an activity assay with leukotoxin and cultured leukemic cells. Results: The results showed a high prevalence (57%) of antibodies against A. actinomycetemcomitans leukotoxin in the analyzed population. These antibodies were correlated with leukotoxin neutralizing capacity as well as with the ELISA titers of A. actinomycetemcomitans-specific IgA and IgG. In addition, high levels of leukotoxin antibodies were correlated with increasing age, but not with periodontal disease parameters or cardiovascular risk factors. Conclusion: Systemic antibodies against A. actionmycetemcomitans leukotoxin were common in this adult Swedish population. These antibodies might contribute to limit the systemic effects of the infection.
INTRODUCTION: The capacity of dentin and collagen to bind DNA and protect against spontaneous and nuclease-induced degradation was evaluated individually and by the incubation of DNA with nuclease-producing bacteria in a mixed culture.
METHODS: Extracted Fusobacterium nucleatum DNA was incubated with dentin shavings or collagen for 90 minutes. The DNA-bound substrates were incubated in different media (water, sera, and DNase I) for up to 3 months. Amplifiable DNA was released from dentin using EDTA,or from collagen using proteinase K, and evaluated by polymerase chain reaction (PCR). The stability of dentin-bound DNA was also assessed in a mixed culture (Parvimonas micra and Pseudoramibacter alactolyticus) containing a DNase-producing species, Prevotella intermedia. Samples were analyzed for amplifiable DNA.
RESULTS: In water, dentin-bound DNA was recoverable by PCR at 3 months compared with no detectable DNA after 4 weeks in controls (no dentin). DNA bound to collagen was detectable by PCR after 3 months of incubation in water. In 10% human sera, amplifiable DNA was detectable at 3 months when dentin bound and in controls (no dentin). In mixed bacterial culture, dentin-bound DNA was recoverable throughout the experimental period (3 months), compared with no recoverable F. nucleatum DNA within 24 hours in controls (no dentin).
CONCLUSIONS: There is a strong binding affinity between DNA and dentin, and between DNA and serum proteins or collagen. These substrates preserve DNA against natural decomposition and protect DNA from nuclease activity, factors that may confound molecular analysis of the endodontic microbiota yet favor paleomicrobiological studies of ancient DNA.
AIM: To study antibody responses associated with molecular mimicry in periodontitis.
MATERIAL AND METHODS: Fifty-four periodontitis cases (mean age 54.0 years) and 44 controls (53.6 years) were examined, after which cases received periodontal treatment. Established immunoassays were used to analyse levels of antibodies against two pathogens, Aggregatibacter actinomycetemcomitans (Aa) and Porphyromonas gingivalis (Pg), heat shock proteins (Hsp), Hsp60, Hsp65, and Hsp70, and epitopes of oxidized low density lipoprotein (oxLDL) (CuOx-LDL and MDA-LDL) in plasma samples that were collected at baseline, after 3 (n=48) and 6 (n=30) months.
RESULTS: When age, sex, smoking habit, and the number of teeth were considered in multivariate logistic regressions, Aa and Pg IgG, Hsp65-IgA, CuOx-LDL-IgG and -IgM and MDA-LDL-IgG antibody levels were associated with periodontitis, whereas Hsp60-IgG2 antibody levels were inversely associated. The Aa antibody levels significantly correlated with the levels of IgA antibodies to Hsp65 and Hsp70, and both OxLDL IgA-antibody levels. The levels of antibodies to Pg correlated with IgG antibodies to Hsp60, Hsp70 and both oxLDL antibody epitopes. None of the antibody levels changed significantly after treatment.
CONCLUSIONS: Periodontitis is associated with persistently high levels of circulating antibodies that are reactive with pathogen- and host-derived antigens.
Clinical intervention studies and experimental studies with lignan-rich diets suggest that lignans may have inhibitory effects on prostate cancer, but no clinical or experimental studies with purified lignans have been published. The purpose of this study was to investigate the effect of a plant lignan 7-hydroxymatairesinol (HMR) on LNCaP human prostate cancer xenografts in athymic mice. Athymic nude male mice were injected subcutaneously with LNCaP cells. Starting 3 days after tumor cell injections, a control diet or a control diet supplemented with 0.15% or 0.30% of HMR was administered to mice and the tumor take rate and growth was observed for 9 weeks. HMR diet inhibited the growth of LNCaP tumors. Mice treated with HMR had smaller tumor volume, lower tumor take rate, increased proportion of nongrowing tumors, and higher tumor cell apoptotic index compared with controls. Furthermore, the cell proliferation index was reduced in mice receiving the 0.30% HMR diet compared with mice receiving the control diet. Our results suggest that dietary HMR started at the early phase of the tumor development inhibits the growth of the LNCaP human prostate cancer xenografts in athymic male mice.
Patients with Kostmann syndrome (severe congenital neutropenia [SCN]) typically normalize their absolute neutrophil count (ANC) upon granulocyte colony-stimulating factor (G-CSF) therapy. However, although they no longer experience life-threatening bacterial infections, they frequently still have recurrent gingivitis and even severe periodontitis, often starting in early childhood. METHODS: We studied the periodontal disease in the four surviving patients belonging to the family originally described by Kostmann. Their odontological records, x-rays, color photos, bacterial cultures, serum antibodies to oral bacteria, and histopathological examinations were reviewed. The data were also correlated to previous investigations on their antibacterial peptides and molecular biology. RESULTS: Three patients had periodontal disease, despite normal ANC and professional dental care, and had neutrophils deficient in antibacterial peptides. One of these patients also had a heterozygous mutation in the neutrophil elastase gene, had severe periodontal disease and overgrowth of the periodontal pathogen Actinobacillus actinomycetemcomitans in the dental flora, and 15 permanent teeth had been extracted by the age of 27. One bone marrow-transplanted patient had no periodontal disease. CONCLUSIONS: Normalized ANC levels are not sufficient to maintain normal oral health in SCN patients, and because neutrophils are important for first-line defense and innate immunity, the deficiency of the antibacterial peptide LL-37 probably explains their chronic periodontal disease. Professional dental care is still important for SCN patients, despite treatment with G-CSF and normal ANC levels. Whether antibacterial peptides play a role in the pathogenesis of periodontitis in other patients remains to be elucidated.
We recently reported that differential gene expression and DNA methylation profiles in blood leukocytes of apparently healthy smokers predicts with remarkable efficiency diseases and conditions known to be causally associated with smoking, suggesting that blood-based omic profiling of human populations may be useful for linking environmental exposures to potential health effects. Here we report on the sex-specific effects of tobacco smoking on transcriptomic and epigenetic features derived from genome-wide profiling in white blood cells, identifying 26 expression probes and 92 CpG sites, almost all of which are affected only in female smokers. Strikingly, these features relate to numerous genes with a key role in the pathogenesis of cardiovascular disease, especially thrombin signaling, including the thrombin receptors on platelets F2R (coagulation factor II (thrombin) receptor; PAR1) and GP5 (glycoprotein 5), as well as HMOX1 (haem oxygenase 1) and BCL2L1 (BCL2-like 1) which are involved in protection against oxidative stress and apoptosis, respectively. These results are in concordance with epidemiological evidence of higher female susceptibility to tobacco-induced cardiovascular disease and underline the potential of blood-based omic profiling in hazard and risk assessment.
The expression of the leukotoxin of Aggregatibacter actinomycetemcomitans is regulated by the leukotoxin promoter. A 530-bp deletion or an 886-bp insertion sequence (IS) element in this region has earlier been described in highly leukotoxic isolates. Here, we report on highly leukotoxic isolate with a 640-bp deletion, which was detected in an adolescent of Ethiopian origin.
Background: The presence of Aggregatibacter actinomycetemcomitans in patients with periodontitis has been extensively studied for decades. Objective: To study the prevalence of A. actinomycetemcomitans in younger and older periodontitis patients and to genetically characterize isolates of this bacterium. Design: Data from microbiological analyses of 3459 subgingival plaque samples collected from 1445 patients, 337 'younger' patients (<= 35 yrs) and 1108 'older' patients (>35 yrs) during 15 years (2000-2014), has been summerized. Isolates of A. actinomycetemcomitans were serotyped, leukotoxin promoter typed (JP2 and non JP2) and arbitrarily primed PCR (APPCR) genotyped. The origin of the JP2 genotype detected in the study population was determined. Results: The prevalence of A. actinomycetemcomitans was higher among younger than older patients and samples from the younger patients contained higher proportions of the bacterium. Serotype b was more prevalent among younger patients and the majorty of these isolates was from the same AP-PCR genotype. The JP2 genotype was detected in 1.2% of the patients, and the majority of these carriers were of non-African origin. Conslusions: For presence and charcteristics of A. actinomycetemcomitans in clinical samples the age of the carriers were a discriminating factor. Additional, apparently non- African carriers of the JP2 genotype of A. actinomycetemcomitans were identified.
The lysosomal enzyme elastase was earlier shown to cleave the complement molecule C3. During somepreliminary experiments on the interactions of certain pathogenic bacteria with the innate defencemechanisms, we observed C3 cleavage, in the presence of elastase, to fragments not previouslydescribed. To elucidate this proteolytic reaction, the present study was conducted. Degradation of C3in mixtures with elastase or cathepsin G was detected by an immunoblot procedure using anti-C3c andanti-C3d antibodies after separating the proteins by SDS-PAGE. Certain C3 fragments were analysedfor amino acid sequence. The results revealed the existence of a cleavage site for elastase at the positionalanine1350 ⁄ lysine1351 of the C3 molecule, which has not been previously described. The fragmentresulted from this cleavage has a size of about 39 kDa and it contains a part or the whole of C3d. Thiscleavage was distinct from the one previously described at position 987 ⁄ 988, which gives a 34 kDaC3d-containing fragment.
Background: Carriers of the JP2 clone of Aggregatibacter actinomycetemcomitans exhibit an enhanced risk for developing aggressive periodontitis compared with individuals carrying non-JP2 clones. While the JP2 clone is almost exclusively detected among adolescents of African descent, reports on Caucasians colonized with the JP2 clone are remarkably few.
Objective: The aim of this paper is to report on the history of periodontal disease and microbiological findings in a Caucasian family.
Material and Methods: A. actinomycetemcomitans and other periodontitis-associated bacterial species in subgingival plaque samples were quantified by conventional culture technique. Leucotoxin promoter typing, serotyping and further characterizations of A. actinomycetemcomitans isolates were performed by PCR. DNA sequencing of the pseudogene, hbpA was performed to determine the origin of the detected JP2 clones. Further, genetically ancestry testing of family members was carried out.
Results: The JP2 clone was detected in samples from two of the family members, a 33-year-old daughter and her 62-year-old mother. Relationship of their JP2 clones with JP2 clone strains from the Mediterranean area of Africa was indicated. Genotyping confirmed the Caucasian origin of all family members.
Conclusions: Caucasian JP2 carriers exist and older subjects can carry the JP2 clone of A. actinomycetemcomitans.
OBJECTIVE: This study describes subgingival bacterial profiles associated with clinical periodontal status in Ghanaian adolescents with or without progression of attachment loss.
MATERIALS AND METHODS: Among 500 adolescents included in a cohort study, 397 returned 2 years later for a periodontal re-examination, including full-mouth CAL measurements. At follow-up, a subgroup of 98 adolescents was also subjected to bacterial sampling with paper points at four periodontal sites (mesial aspect of 11, 26, 31, and 46) and analyzed with the checkerboard DNA-DNA hybridization technique against DNA-probes from nine periodontitis-associated bacterial species.
RESULTS: The 98 Ghanaian adolescents examined in the present study were similar to the entire group examined at the 2-year follow-up with respect to age, gender, and CAL ≥3 mm. A high detection frequency of Fusobacterium nucleatum and Prevotella intermedia (>99%) using checkerboard analysis was found, while for Aggregatibacter actinomycetemcomitans the detection frequency was <50%. A strong correlation was found at the individual level between the presence of P. intermedia and the total CAL change, and P. intermedia and Porphyromonas gingivalis were strongly correlated with a change in CAL and probing pocket depth (PPD) at the sampled sites. In a linear regression model, a significant discriminating factor for the total CAL change in the dentition during the 2-year follow-up period was obtained for P. intermedia and public school.
CONCLUSION: This study indicates that subgingival bacterial species other than A. actinomycetemcomitans, for example, P. intermedia, have a significant association with periodontal breakdown (change in CAL) in Ghanaian adolescents with progression of periodontal attachment loss.
BACKGROUND: A relationship between rheumatoid arthritis (RA) and periodontitis has been suggested from findings that individuals with RA are prone to have advanced periodontitis and vice versa. In search of possible common pathogenetic features of these two diseases, we investigated the presence of citrullinated proteins and expression of endogenous peptidylarginine deiminases (PAD2 and PAD4), in periodontal tissue of individuals with periodontitis and healthy controls, in relation to the periodontal pathogens Porphyromonas gingivalis (P. gingivalis) and Aggregatibacter actinomycetemcomitans (A. actinomycetemcomitans), producing leukotoxin as virulence factor. These two oral bacteria have been suggested to be linked to anti-citrullinated protein antibodies in patients with RA.
METHODS: Gingival tissue biopsies were obtained from 15 patients with periodontitis and 15 individuals without periodontal disease. Presence of CD3-positive lymphocytes, citrullinated proteins, PAD2, PAD4, P. gingivalis as well as A. actinomycetemcomitans and Mannheimia haemolytica produced leukotoxins were analysed by immunohistochemistry, followed by triple-blind semi-quantitative analysis. Mann-Whitney and Fisher's exact tests were used to analyse differences between groups. PADI2 and PADI4 mRNA levels were assessed by RT-qPCR and analysed using Wilcoxon signed rank test.
RESULTS: Increased staining of citrullinated proteins was observed in gingival connective tissue from subjects with periodontitis (80%, 12/15) compared to healthy gingival tissue (27%, 4/15), whereas no differences were observed in gingival epithelium. There was also an increased staining of the citrullinating enzymes PAD2 and PAD4 in gingival connective tissue of patients with periodontitis whereas similar levels of PAD2 and PAD4 were observed in the gingival epithelium of the two groups. Similarly, the mRNA levels of PADI2 and PADI4 were also increased in the gingival tissue of patients with periodontitis compared to healthy controls. Furthermore, presence of P. gingivalis and leukotoxins was comparable in both epithelium and connective tissue, from the different investigated individuals with and without periodontitis, and there were no correlations between the presence of periodontal pathogens and the expression of citrullinated proteins or PAD enzymes.
CONCLUSION: Chronic gingival inflammation is associated with increased local citrullination and PAD2 and PAD4 expression in periodontitis. The increased citrullination and PAD2 and PAD4 expression in periodontitis were, however, independent of the presence of periodontal pathogen P. gingivalis and A. actinomycetemcomitans leukotoxin.
It has previously been shown that the presence of Aggregatibacter actinomycetemcomitans in subgingival plaque is significantly associated with increased risk for clinical attachment loss. The highly leukotoxic JP2 genotype of this bacterium is frequently detected in adolescents with aggressive forms of periodontitis. The aims of the study were to quantify the levels of JP2 and non-JP2 genotypes of A. actinomycetemcomitans in saliva of Moroccan adolescents with the JP2 genotype earlier detected in the subgingival plaque. The salivary concentrations of inflammatory proteins were quantified and linked to the clinical parameters and microbial findings. Finally, a mouth rinse with leukotoxin-neutralizing effect was administrated and its effect on the levels the biomarkers and A. actinomycetemcomitans examined. The study population consisted of 22 adolescents that previously were found to be positive for the JP2 genotype in subgingival plaque. Periodontal registration and sampling of stimulated saliva was performed at baseline. A mouth rinse (active/placebo) was administrated, and saliva sampling repeated after 2 and 4 weeks rinse. The salivary levels of JP2 and non-JP2 were analyzed by quantitative PCR and inflammatory proteins by ELISA. Both the JP2 and the non-JP2 genotype were detected in all individuals with significantly higher levels of the non-JP2. Enhanced levels of the JP2 genotype of A. actinomycetemcomitans was significantly correlated to the presence of attachment loss (≥3 mm). Salivary concentrations of inflammatory biomarkers did not correlate to periodontal condition or levels of A. actinomycetemcomitans. The use of active or placebo leukotoxin-neutralizing mouth rinse did not significantly interfered with the levels of these biomarkers. Saliva is an excellent source for detection of A. actinomycetemcomitans on individual basis, and high levels of the JP2 genotype were significantly associated with the presence of clinical attachment loss.
PCBs are classified as xenoestrogens and carcinogens and their health risks may be sex-specific. To identify potential sex-specific responses to PCB-exposure we established gene expression profiles in a population study subdivided into females and males. Gene expression profiles were determined in a study population consisting of 512 subjects from the EnviroGenomarkers project, 217 subjects who developed lymphoma and 295 controls were selected in later life. We ran linear mixed models in order to find associations between gene expression and exposure to PCBs, while correcting for confounders, in particular distribution of white blood cells (WBC), as well as random effects. The analysis was subdivided according to sex and development of lymphoma in later life. The changes in gene expression as a result of exposure to the six studied PCB congeners were sex- and WBC type specific. The relatively large number of genes that are significantly associated with PCB-exposure in the female subpopulation already indicates different biological response mechanisms to PCBs between the two sexes. The interaction analysis between different PCBs and WBCs provides only a small overlap between sexes. In males, cancer-related pathways and in females immune system-related pathways are identified in association with PCBs and WBCs. Future lymphoma cases and controls for both sexes show different responses to the interaction of PCBs with WBCs, suggesting a role of the immune system in PCB-related cancer development.
Experimental and epidemiological data suggest that vitamin D play a role in pathogenesis and progression of cancer, but prospective data on head and neck cancer (HNC) and oesophagus cancer are limited. The European Prospective Investigation into Cancer and Nutrition (EPIC) study recruited 385,747 participants with blood samples between 1992 and 2000. This analysis includes 497 case-control pairs of the head and neck and oesophagus, as well as 443 additional controls. Circulating 25(OH)D3 were measured in pre-diagnostic samples and evaluated in relation to HNC and oesophagus cancer risk and post-diagnosis all-cause mortality. After controlling for risk factors, a doubling of 25(OH)D3 was associated with 30% lower odds of HNC (OR 0.70, 95% confidence interval [95% CI] 0.56-0.88, Ptrend = 0.001). Subsequent analyses by anatomical sub-site indicated clear inverse associations with risk of larynx and hypopharynx cancer combined (OR 0.55, 95CI% 0.39-0.78) and oral cavity cancer (OR 0.60, 95CI% 0.42-0.87). Low 25(OH)D3 concentrations were also associated with higher risk of death from any cause among HNC cases. No clear association was seen with risk or survival for oesophageal cancer. Study participants with elevated circulating concentrations of 25(OH)D3 had decreased risk of HNC, as well as improved survival following diagnosis.
The utility of blood-based omic profiles for linking environmental exposures to their potential health effects was evaluated in 649 individuals, drawn from the general population, in relation to tobacco smoking, an exposure with well-characterised health effects. Using disease connectivity analysis, we found that the combination of smoking-modified, genome-wide gene (including miRNA) expression and DNA methylation profiles predicts with remarkable reliability most diseases and conditions independently known to be causally associated with smoking (indicative estimates of sensitivity and positive predictive value 94% and 84%, respectively). Bioinformatics analysis reveals the importance of a small number of smoking-modified, master-regulatory genes and suggest a central role for altered ubiquitination. The smoking-induced gene expression profiles overlap significantly with profiles present in blood cells of patients with lung cancer or coronary heart disease, diseases strongly associated with tobacco smoking. These results provide proof-of-principle support to the suggestion that omic profiling in peripheral blood has the potential of identifying early, disease-related perturbations caused by toxic exposures and may be a useful tool in hazard and risk assessment.
OBJECTIVE: In this study, we analyzed the production of Interleukin-1 beta (IL-1β) and IL-6 by activated CD4+ cells obtained from aggressive periodontitis (AgP) patients in comparison with healthy subjects (HC).
MATERIALS AND METHODS: CD4+ cells were automatically separated from lymphocytes obtained from peripheral blood of patients with AgP and healthy controls. Cells were activated for 4, 8, and 24 h with three different stimuli: anti-CD3/anti-CD28, phytohemagglutinin (PHA), and Porphyromonas gingivalis (P. gingivalis) outer membrane protein (OMP). Protein levels were measured in supernatants of activated CD4+ cells by a bead-based immunoassay (CBA). In addition, serum antibodies against P. gingivalis were determined. Data were analyzed using U test (p < 0.05).
RESULTS: T helper cells of AgP patients activated with P. gingivalis OMP produced higher levels of IL-1β and IL-6 in comparison with healthy controls (p < 0.05). Neither the activation with anti-CD3/anti-CD28 nor the activation with PHA showed significantly different production of IL-1β and IL-6 by the cells 25 % of patients and 17 % of controls presented with high serum reactivity to P. gingivalis.
CONCLUSION: In view of these results, it is possible to conclude that P. gingivalis contributes to the pathogenesis of AgP by inducing high levels of pro-inflammatory cytokines such as IL-1β and IL-6 by peripheral CD4+ T helper cells.
CLINICAL RELEVANCE: In accordance with the clinical parameters and the immunological data, we suggest that full-mouth disinfection with adjunctive systemic antibiotics might be the anti-infectious non-surgical periodontal treatment of choice in this type of patients. Microbiological analyses at the beginning and at the end of the periodontal treatment are recommended. However, it is necessary to verify these data in longitudinal clinical studies.
The purpose of this paper is, first, to describe the organization, sampling procedures, availability of samples/database, ethical considerations, and quality control program of the Northern Sweden Health and Disease Study Cohort. Secondly, some examples are given of studies on cardiovascular disease and diabetes with a focus on the biomarker programme. The cohort has been positioned as a national and international resource for scientific research.
Rye bran contains a high content not only of dietary fibre, but also of plant lignans and other bioactive compounds in the so-called dietary fibre complex. Blood concentrations of lignans such as enterolactone have been used as biomarkers of intake of lignan-rich plant food. At present,evidence from studies in human subjects does not warrant the conclusion that rye, whole grains orphyto-oestrogens protect against cancer. Some studies, however, have pointed in that direction,especially in relation to cancers of the upper digestive tract. A number of prospective epidemiological studies have clearly shown a protective effect of wholegrain cereals against myocardial infarctions. A corresponding protective effect against diabetes and ischaemic stroke(brain infarct) has also been demonstrated. It seems reasonable to assume that these protective effects are associated with one or more factors in the dietary fibre complex.
For decades, Aggregatibacter actinomycetemcomitans has been associated with aggressive forms of periodontitis in adolescents. In the middle of the 1990s, a specific JP2 clone of A. actinomycetemcomitans, belonging to the cluster of serotype b strains of A. actinomycetemcomitans and having a number of other characteristics, was found to be strongly associated with aggressive forms of periodontitis, particularly in North Africa. Although several longitudinal studies still point to the bacterial species, A. actinomycetemcomitans as a risk factor of aggressive periodontitis, it is now also widely accepted that the highly leukotoxic JP2 clone of A. actinomycetemcomitans is implicated in rapidly progressing forms of aggressive periodontitis. The JP2 clone strains are highly prevalent in human populations living in Northern and Western parts of Africa. These strains are also prevalent in geographically widespread populations that have originated from the Northwest Africa. Only sporadic signs of a dissemination of the JP2 clone strains to non-African populations have been found despite Africans living geographically widespread for hundreds of years. It remains an unanswered question if a particular host tropism exists as a possible explanation for the frequent colonization of the Northwest African population with the JP2 clone. Two exotoxins of A. actinomycetemcomitans are known, leukotoxin (LtxA) and cytolethal distending toxin (Cdt). LtxA is able to kill human immune cells, and Cdt can block cell cycle progression in eukaryotic cells and thus induce cell cycle arrest. Whereas the leukotoxin production is enhanced in JP2 clone strains thus increasing the virulence potential of A. actinomycetemcomitans, it has not been possible so far to demonstrate such a role for Cdt. Lines of evidence have led to the understanding of the highly leukotoxic JP2 clone of A. actinomycetemcomitans as an aetiological factor of aggressive periodontitis. Patients, who are colonized with the JP2 clone, are likely to share this clone with several family members because the clone is transmitted through close contacts. This is a challenge to the clinicians. The patients need intense monitoring of their periodontal status as the risk for developing severely progressing periodontal lesions are relatively high. Furthermore, timely periodontal treatment, in some cases including periodontal surgery supplemented by the use of antibiotics, is warranted. Preferably, periodontal attachment loss should be prevented by early detection of the JP2 clone of A. actinomycetemcomitans by microbial diagnostic testing and/or by preventive means.
BACKGROUND: Aggressive periodontitis is associated with the presence of Aggregatibacter actinomycetemcomitans, a leukotoxin (Ltx)-producing periodontal pathogen. Ltx has the ability to lyse white blood cells including neutrophils.
OBJECTIVES: This study was aimed at investigating the interactions between neutrophils and Ltx with regard to the chemotactic properties of Ltx and the release of neutrophil extracellular traps (NETs).
METHODS: Neutrophils from healthy blood donors were isolated and incubated for 30 min and 3 h with increasing concentrations of Ltx (1, 10, and 100 ng/mL) as well as with A. actinomycetemcomitans strains (NCTC 9710 and HK 1651) producing different levels of Ltx. Formation of NETs and cell lysis were assessed by microscopy, fluorescence-based assays, and measurement of released lactate dehydrogenase. Neutrophil migration in response to different Ltx gradients was monitored by real-time video microscopy, and image analysis was performed using ImageJ software.
RESULTS: Although Ltx (10 and 100 ng/mL) and the leukotoxic A. actinomycetemcomitans strain HK 1651 lysed some neutrophils, other cells were still capable of performing NETosis in a concentration-dependent manner. Low doses of Ltx and the weakly leukotoxic strain NCTC 9710 did not lead to neutrophil lysis, but did induce some NETosis. Furthermore, all three concentrations of Ltx enhanced random neutrophil movement; however, low directional accuracy was observed compared with the positive control (fMLP).
CONCLUSIONS: The results indicate that Ltx acts both as a neutrophil activator and also causes cell death. In addition, Ltx directly induces NETosis in neutrophils prior to cell lysis. In future studies, the underlying pathways involved in Ltx-meditated neutrophil activation and NETosis need to be investigated further.
Background and Objectives: The cytolethal distending toxin (Cdt) is a highly conserved exotoxin that are produced by a number of Gram negative bacteria, including Aggregatibacter actinomycetemcomitans, and affects mammalian cells by inhibiting cell division and causing apoptosis. A complete cdt-operon is present in the majority of A. actinomycetemcomitans, but the proportion of isolates that lack cdt-encoding genes (A, B and C) varies according to the population studied. The objectives of this study were to examine serotype, Cdt-genotype, and Cdt-activity in isolates of A. actinomycetemcomitans collected from an adolescent West African population and to examine the association between the carrier status of A. actinomycetemcomitans and the progression of attachment loss (AL).
Material and Methods: A total of 249 A. actinomycetemcomitans isolates from 200 Ghanaian adolescents were examined for serotype and cdt-genotype by PCR. The activity of the Cdt-toxin was examined by DNA-staining of exposed cultured cells and documented with flow cytometry. The periodontal status of the participants was examined at baseline and at a two-year follow-up.
Results: Presence of all three cdt-encoding genes was detected in 79% of the examined A. actinomycetemcomitans isolates. All these isolates showed a substantial Cdt-activity. The two different cdt-genotypes (with and without presence of all three cdt-encoding genes) showed a serotype-dependent distribution pattern. Presence of A. actinomycetemcomitans was significantly associated with progression of AL (OR = 5.126; 95% CI = [2.994 - 8.779], p < 0.001).
Conclusion: A. actinomycetemcomitans isolated from the Ghanaian adolescents showed a distribution of serotype and cdt-genotype in line with results based on other previously studied populations. Presence of A. actinomycetemcomitans was significantly associated with disease progression, in particular the b serotype, whereas the association with disease progression was not particularly related to cdt-genotype, and Cdt-activity.
Aggregatibacter actinomycetemcomitans is a Gram-negative periodontitis-associated bacterium that expresses a toxin that selectively affects leukocytes. This leukotoxin is encoded by an operon belonging to the core genome of this bacterial species. Variations in the expression of the leukotoxin have been reported, and a well-characterized specific clonal type (JP2) of this bacterium with enhanced leukotoxin expression has been isolated. In particular, the presence of the JP2 genotype significantly increases the risk for the progression of periodontal attachment loss (AL). Based on these findings we hypothesized that variations in the leukotoxicity are linked to disease progression in infected individuals. In the present study, the leukotoxicity of 239 clinical isolates of A. actinomycetemcomitans was analysed with different bioassays, and the genetic peculiarities of the isolates were related to their leukotoxicity based on examination with molecular techniques. The periodontal status of the individuals sampled for the presence of A. actinomycetemcomitans was examined longitudinally, and the importance of the observed variations in leukotoxicity was evaluated in relation to disease progression. Our data show that high leukotoxicity correlates with an enhanced risk for the progression of AL. The JP2 genotype isolates were all highly leukotoxic, while the isolates with an intact leukotoxin promoter (non-JP2 genotypes) showed substantial variation in leukotoxicity. Genetic characterization of the non-JP2 genotype isolates indicated the presence of highly leukotoxic genotypes of serotype b with similarities to the JP2 genotype. Based on these results, we conclude that A. actinomycetemcomitans harbours other highly virulent genotypes besides the previously described JP2 genotype. In addition, the results from the present study further highlight the importance of the leukotoxin as a key virulence factor in aggressive forms of periodontitis.
Periodontitis is an infection-induced inflammatory disease that causes loss of the tooth supporting tissues. Much focus has been put on comparison of the microbial biofilm in the healthy periodontium with the diseased one. The information arising from such studies is limited due to difficulties to compare the microbial composition in these two completely different ecological niches. A few longitudinal studies have contributed with information that makes it possible to predict which individuals who might have an increased risk of developing aggressive forms of periodontitis, and the predictors are either microbial or/and host-derived factors. The most conspicuous condition that is associated with disease risk is the presence of Aggregatibacter actinomycetemcomitans at the individual level. This Gram-negative bacterium has a great genetic variation with a number of virulence factors. In this review we focus in particular on the leukotoxin that, based on resent knowledge, might be one of the most important virulence factors of A. actinomycetemcomitans.
AIM: To assess the progression of attachment loss (AL) during a two-year period according to the presence of JP2 and non-JP2 genotypes of A. actinomycetemcomitans in a Ghanaian adolescent population.
METHODS: A total of 500 adolescents (mean age 13.2 years, SD± 1.5) were enrolled in the study. After two years, 397 (79.4%) returned for a periodontal re-examination, including the measurement of AL. The carrier status of the JP2 and non-JP2 genotypes of A. actinomycetemcomitans was evaluated in a baseline examination two years earlier.
RESULTS: Individuals who carried the JP2 genotype of A. actinomycetemcomitans had a significantly increased risk (relative risk (RR) = 7.3) of developing AL ≥ 3 mm. The mean AL at the follow-up and the mean two-year progression of AL was significantly higher in the JP2 genotype-positive group (n=38) compared with the group positive for the non-JP2 genotypes of A. actinomycetemcomitans (n=169), and the group of A. actinomycetemcomitans-negative individuals (n=190). The JP2 genotype was strongly associated with the progression of AL ≥ 3 mm (OR= 14.3). The non-JP2 genotypes of A. actinomycetemcomitans were also, however less pronounced, associated with the progression of AL ≥ 3 mm (OR=3.4).
CONCLUSION: The JP2 genotype of A. actinomycetemcomitans is strongly associated with the progression of AL.
Background: Limited data are reported concerning the presence of A. actinomycetemcomitans and attachment loss (AL) in sub-Saharan countries. The authors investigate the carrier frequency of JP2 and non-JP2 genotypes of A. actinomycetemcomitans and the presence of AL in Ghanaian adolescents and evaluate socioeconomic conditions and oral hygiene practices. Methods: Five hundred individuals (mean +/- SD age: 13.2 +/- 1.5 years) in public and private schools were interviewed about demographic characteristics and oral hygiene practices and were given a full-mouth periodontal examination. Subgingival plaque samples were obtained from periodontal sites around permanent first molars and incisors. The carrier status of A. actinomycetemcomitans at the individual level was determined based on results obtained by cultivation and polymerase chain reaction. Results: The findings of this study show a relatively high carrier rate of JP2 and non-JP2 genotypes of Aggregatibacter actinomycetemcomitans in the Ghanaian adolescent population and the presence of this bacterium is associated with the occurrence of AL. The overall carrier rate of A. actinomycetemcomitans was 54.4%, and the highly leukotoxic JP2 genotype was detected in 8.8% of the study population. A total of 107 (21.4%) individuals had >= 1 tooth with AL >= 3 mm. The majority of the individuals carrying A. actinomycetemcomitans (80.1%) (P<0.001) and of the periodontally diseased individuals (91.6%) (P<0.001) were found in public schools. Conclusions: A. actinomycetemcomitans and AL were frequently found in Ghanaian adolescents. The school type was the strongest predictor of both presence of A. actinomycetemcomitans and AL.
Apoptotic cysteine-aspartate proteases (caspases) are essential for the progression and execution of apoptosis, and detection of caspase fragmentation or activity is often used as markers of apoptosis. Cisplatin (cis-diamminedichloroplatinum (II)) is a chemotherapeutic drug that is clinically used for the treatment of solid tumours. We compared a cisplatin-resistant pleural malignant mesothelioma cell line (P31res1.2) with its parental cell line (P31) regarding the consequences of in vitro acquired cisplatin-resistance on basal and cisplatin-induced (equitoxic and equiapoptotic cisplatin concentrations) caspase-3, -8 and -9 fragmentation and proteolytic activity. Acquisition of cisplatin-resistance resulted in basal fragmentation of caspase-8 and -9 without a concomitant increase in proteolytic activity, and there was an increased basal caspase-3/7 activity. Similarly, cisplatin-resistant non-small-cell lung cancer cells, H1299res, had increased caspase-3 and -9 content compared with the parental H1299 cells. In P31 cells, cisplatin exposure resulted in caspase-9-mediated caspase-3/7 activation, but in P31res1.2 cells the cisplatin-induced caspase-3/7 activation occurred before caspase-8 or -9 activation. We therefore concluded that in vitro acquisition of cisplatin-resistance rendered P31res1.2 cells resistant to caspase-8 and caspase-9 fragments and that cisplatin-induced, initiator-caspase independent caspase-3/7 activation was necessary to overcome this resistance. Finally, the results demonstrated that detection of cleaved caspase fragments alone might be insufficient as a marker of caspase activity and ensuing apoptosis induction.
Despite declining incidence rates, gastric cancer (GC) is a major cause of death worldwide. Its aetiology may involve dietary antioxidant micronutrients such as carotenoids and tocopherols. The objective of this study was to determine the association of plasma levels of seven common carotenoids, their total plasma concentration, retinol and - and -tocopherol, with the risk of gastric adenocarcinoma in a case-control study nested within the European Prospective Investigation into Cancer and Nutrition (EPIC), a large cohort involving 10 countries. A secondary objective was to determine the association of total sum of carotenoids, retinol and -tocopherol on GCs by anatomical subsite (cardia/noncardia) and histological subtype (diffuse/intestinal). Analytes were measured by high-performance liquid chromatography in prediagnostic plasma from 244 GC cases and 645 controls matched by age, gender, study centre and date of blood donation. Conditional logistic regression models adjusted by body mass index, total energy intake, smoking and Helicobacter pylori infection status were used to estimate relative cancer risks. After an average 3.2 years of follow-up, a negative association with GC risk was observed in the highest vs the lowest quartiles of plasma -cryptoxanthin (odds ratio (OR)=0.53, 95% confidence intervals (CI)=0.30-0.94, Ptrend=0.006), zeaxanthin (OR=0.39, 95% CI=0.22-0.69, Ptrend=0.005), retinol (OR=0.55, 95% CI=0.33-0.93, Ptrend=0.005) and lipid-unadjusted -tocopherol (OR=0.59, 95% CI=0.37-0.94, Ptrend=0.022). For all analytes, no heterogeneity of risk estimates or significant associations were observed by anatomical subsite. In the diffuse histological subtype, an inverse association was observed with the highest vs lowest quartile of lipid-unadjusted -tocopherol (OR=0.26, 95% CI=0.11-0.65, Ptrend=0.003). These results show that higher plasma concentrations of some carotenoids, retinol and -tocopherol are associated with reduced risk of GC.
AIM: The present study aims to determine the susceptibility of Aggregatibacter actinomycetemcomitans to amoxicillin by investigating a large collection of oral strains of diverse geographical origin.
METHODS: Two hundred and fifty-seven A. actinomycetemcomitans strains were serotyped using a multiplex polymerase chain reaction, and minimal inhibitory concentration (MIC) values of amoxicillin were determined using the agar dilution method (range 0.25 to 8.0 mg/L). The plates were spot-wise inoculated with approximately 104 colony-forming units, incubated in 5% CO2 at 37 C°, and visually inspected after 24 and 48 hours. A MIC ≤ 2.00 mg/L was categorised as susceptible using EUCAST interpretative criteria for Haemophilus species.
RESULTS: Amoxicillin MIC-values varied from 0.25 mg/L to 2.00 mg/L, and all tested strains, including strains previously reported as resistant, were susceptible to amoxicillin. The MIC50 was 1.00 mg/L and the MIC90 was 2.00 mg/L.
CONCLUSION: Meticulous investigation of strains including isolates previously reported as resistant could not confirm the emergence of resistance to β-lactams in A. actinomycetemcomitans. Based on the present in vitro results, amoxicillin can be considered a key oral antimicrobial agent for treatment of A. actinomycetemcomitans. This article is protected by copyright. All rights reserved.
Leukotoxic potential of Aggregatibacter actinomycetemcomitans strains has been studied by the use of several methods, and results differ depending on the methods used. The aim of the present study was to perform a comprehensive examination of the leukotoxic potential of a collection of A. actinomycetemcomitans strains by use of three quantitative methods, Western blotting, ELISA, and mRNA expression assay and compare these results with previous data obtained by a cell lysis assay. A higher leukotoxic potential among JP2 genotype strains compared to non-JP2 genotype strains of A. actinomycetemcomitans was found by Western blotting, ELISA and mRNA expression assay. Leukotoxicity as determined by cell lysis assay showed a variation among strains examined, not only depending on being part of JP2 genotype vs. non-JP2 genotype group of A. actinomycetemcomitans. The leukotoxicity of A. actinomycetemcomitans strains as determined by cell lysis assay did not correspond to the leukotoxic potential of A. actinomycetemcomitans strains as determined by three quantitative methods. A comparison of the results obtained by ELISA and mRNA expression assay showed a reasonable correlation between these two methods. It seems important to use more than one method to assess the LtxA-related virulence capacity of A. actinomycetemcomitans in order to obtain comprehensive understanding of the leukotoxic potential of A. actinomycetemcomitans strains.
Aggregatibacter actinomycetemcomitans has been described as a member of the indigenous oral microbiota of humans, and is involved in the pathology of periodontitis and various non-oral infections. This bacterium selectively kills human leukocytes through expression of leukotoxin, a large pore-forming protein that belongs to the Repeat in Toxin (RTX) family. The specificity of the toxin is related to its prerequisite for a specific target cell receptor, LFA-1, which is solely expressed on leukocytes. The leukotoxin causes death of different leukocyte populations in a variety of ways. It activates a rapid release of lysosomal enzymes and MMPs from neutrophils and causes apoptosis in lymphocytes. In the monocytes/macrophages, the toxin activates caspase-1, a cysteine proteinase, which causes a proinflammatory response by the activation and secretion of IL-1β and IL-18. A specific clone (JP2) of A. actinomycetemcomitans with enhanced leukotoxin expression significantly correlates to disease onset in infected individuals. Taken together, the mechanisms by which this toxin kills leukocytes are closely related to the pathogenic mechanisms of inflammatory disorders, such as periodontitis. Therapeutic strategies targeting the cellular and molecular inflammatory host response in periodontal diseases might be a future treatment alternative
Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis secrete several potent virulence factors and are known to be two of the major periodontal pathogens. In the present case-control study, the systemic immunoreactivity to A. actinomycetemcomitans exotoxins, cytolethal distending toxin (Cdt) and leukotoxin was analyzed in adult subjects with periodontitis and in periodontally healthy controls. Furthermore, systemic immunoreactivity to P. gingivalis was analyzed in these subjects. Reactivity to the A. actinomycetemcomitans toxins was determined in bioassays that quantified neutralizing antibodies, and P. gingivalis antibodies were detected by enzyme-linked immunosorbent assay (ELISA). The results showed a significantly enhanced immunoreactivity to P. gingivalis in the subjects with periodontitis, while the reactivity to A. actinomycetemcomitans leukotoxin showed no significant difference between patients and controls. However, combined immunoreactivity to leukotoxin and Cdt was more prevalent in the subjects with periodontitis than in the controls. In addition, immunoreactivity to leukotoxin correlated to periodontitis in men but not in women. In conclusion, data from the present study indicate that immunoreactivity to P. gingivalis is frequent in adult periodontitis, while the role of A. actinomycetemcomitans seems to be more complex and depends on gender of the infected subject as well as the virulence of the bacteria. (c) Eur J Oral Sci, 2005
BACKGROUND: The leukotoxin expressed by Aggregatibacter actinomycetemcomitans is a powerful exotoxin, which can cause imbalance in the host response. Immunoreactivity to the leukotoxin is a marker for the presence of leukotoxic A. actinomycetemcomitan, a presence that may modify the disease pattern of the colonized individuals. The aim of the present study was to examine the presence of systemic immunoreactivity to A. actionmycetemcomitans leukotoxin in relation to clinical and inflammatory findings in individuals with or without periodontitis (n = 88).
METHODS: The periodontal status was examined in a population of cases (n = 49) and controls (n = 39), and the cases received periodontal treatment. Systemic biomarkers associated with inflammation and infections, as well as the clinical parameters, were analyzed at baseline, three months after treatment and six months after.
RESULTS: The presence of immunoreactivity against leukotoxin was associated with impaired remission of the disease after periodontal treatment. This immunoreactivity was also significantly associated with increased systemic levels of A. actinomycetemcomtans-specific immunoglobulins and increasing age.
CONCLUSION: The presence and levels of systemic immunoreactivity against A. actinomycetemcomitans leukotoxin are associated with a decreased remission after otherwise successful periodontal treatment.
The leukotoxin of the periodontopathogen Actinobacillus actinomycetemcomitans is an important virulence factor that lyses human neutrophils and monocytes and thus, it may enable the bacterium to evade the local host defense. The toxin also induces degranulation of neutrophils and cytokine release in monocytes. To trigger these biological activities, leukotoxin has to be released from the bacterium and diffuse into the periodontal tissues. To date, the conditions found to cause toxin release have been artificial and have included high ion concentration and alkaline conditions. To study the release of the toxin under conditions mimicking the natural environment of the periodontium the ability of human serum to enable leukotoxin release from the bacterial surface was examined. Suspensions of leukotoxic A. actinomycetemcomitans strains were incubated with various concentrations of human serum or serum albumin. The suspensions were centrifuged and the leukotoxin in the supernatants or the cell pellets was detected by gel electrophoresis and immunoblotting. Serum was found to cause the rapid release of leukotoxin from the bacteria in a concentration-dependent manner. Pure albumin exhibited a similar effect. The leukotoxin released was active against human neutrophils. Only a minor proportion of it was associated with membranous vesicles produced by the bacteria. The results indicate that serum, a fluid closely related to the exudate in inflamed periodontal pockets, releases leukotoxin from the cell surface of A. actinomycetemcomitans. The process may enable the diffusion of the toxin from the bacterial biofilm into the surrounding tissues, where it can exert its biological effect.


text:
4 more days and I get my online life back.
I think I have learned what I needed to learn. I'll recap:
1. I do not NEED the internet and social networking. Should tragedy befall us and we had to cut things...I'd turn it off, no problem.
2. Although I do not NEED social networking...I WANT social networking. Being a WAHM generally surrounded by humans under 4 feet tall, a dog, no one or a combination of those three....I need the connection sites like facebook bring me. I also, apparently, like to talk and social networking helps me ease that.
3. I did not accomplish many of the goals I wanted to accomplish. I did start making small changes to our diet and grocery purchases. I'm still a coupon fanatic but am seeking out more healthy purchases with the coupons. But in these tight times, I gotta do what I gotta do to feed my family!
4. I did not create much at all. But this has little do with with disconnecting. My kids are home on summer break and all consuming. So summer was probably NOT a great time to try disconnecting.
5. I did see how disconnecting is valuable and am trying to decide what day each week I want to be my "unplugged" day.
here is how my goals broke down. (Mind you these were long term goals listed here, but I wanted to update my progress.) (Updates are in Red)
*Integrate stevia and blue agave nectar into diet more.--Working on! They no longer make Sun Crystals, my sweetener of choice for my coffee, but I can make my own by mixing stevia and real cane sugar. I also got some local honey and agave nectar and have been using them in coffee and smoothies!
*Use up current stockpile of all items that contain high fructose corn syrup, and multiple chemicals on the label. Working on....slowly and surely.
*As we use that stock up, research alternatives for such items.Working on....slowly and surely.
*Limit our soda intake. Once it's used up...purchase natural sodas without HFCS. Working on....slowly and surely.
*Drink more Water. Working on....slowly and surely. Sort of failing in this one...
*Cut back on sugar. Working on....slowly and surely. Sort of failing in this one...
*Eliminate or drastically cut back on food dyes. Working on....slowly and surely. I think I'm doing good in this one, it was mostly for the boys and as I look at their diet, they don't eat much with food dyes. Their fruit jerky (fruit strips) are colored with fruit and veggie dyes. Their cereals don't contain dyes. Their juices are all natural. Probably their biggest downfall is the occasional popsicle at PawPaw's house, but I also want to be realistic, the sometimes popsicle isn't going to kill them. Fruit Chewy Treats are probably the last majorly dyed food they ingest, and again, I'm not going to go all Gestapo on their butts.
*Purchase meats, dairy and produce that I KNOW where it came from. Working on....slowly and surely. This information is harder to get than I thought.
*Purchase red meat and chicken only from our local grocery store Working on....slowly and surely. Local and organic meats are SUPER costly.
*Purchase the 'dirtiest' produce list http://www.kitchenstewardship.com/2010/04/29/2010-dirty-dozen-produce-list-released-by-the-ewg/ organic and preferably, locally grown.Working on....slowly and surely.
*Purchase locally grown and organic fed dairy products. Working on....slowly and surely. Thankfully most of the milk I buy is from local farmers who supposedly don't use hormones or any of that. Still trying to find a good, clean, quiet source of Raw Milk. Since it's illegal I can't even tell you if I find one anyways.
*Exercise more! FAILED miserably. I've walked the dog...that's about it. I've put on about 8 lbs in the past six months. I'm a new mission to get ripped.
*Spend more time in bible study. Working on....slowly and surely. I've not done enough of this, absolutely not.
*Cleaning and Housework. Working on....slowly and surely. Getting better. I'm still trying to make myself get on a good schedule, but I'm not a good schedule keeper. But I've done okay keeping things going, since it is summer and both kids are home...it's a lot harder to keep things clean!
*Purge some of the plastic. Working on....slowly and surely. Purged some and am working on purging more. Slowly (it's costly!) replacing my plastic items with glass.
*Read More! Doing good! I'm up to 9 books finished on my kindle alone. I think my goal to read in 2011 was 20-25. I'm reading a few books off the kindle (what are these called now, In Real Life books?) and as always, my stack of magazines.
*Un-Plug More. Semi Accomplished!! As this girl disconnected series draws to a close, I admit I've sneaked on some social networking sites, I've still used my iPad and I've still done what I needed to do for my business online. But I was offline a WHOLE lot more than I've ever been in recent years. I of course didn't have a ton of time to BE online since it's summer, but not even being ALLOWED online helped ease my stress and guilt for 'not getting enough done.'
So, overall, I'm not doing TOO terrible I think!!
See ya'll soon! FOUR MORE DAYS!
Sunday, June 26, 2011
girl disconnected...4 more days
1 comment:
- Heather said...
4 more days! Woot!
- June 26, 2011 at 8:03 PM
Subscribe to: Post Comments (Atom)

subset: commoncrawl


text:
Most quirky electric car concepts from Japan
Japan is known for both its engineering ingenuity and quirky weirdness and the following concept cars illustrate this dual principle perfectly. Among our favourites is the Eliica (ELectric LIthium-Ion CAr) that not only boasts 8 wheels but an electric motor in each.
Other concepts have more serious backers even, such as the early ideas of Toyota and Mazda. In 1974, Toyota unveiled the RV-2 concept. The rear opened partially and created a space across which you could span a tent.
We also like the Bambgoo. Developed by Kyoto University, the Bambgoo was a small electric car made from nothing than bamboo and also capable of running 31 miles on a single charge.


text:
Catholic Family Service (CFS) has been officially awarded a four-year accreditation through the Canadian Centre for Accreditation (CCA).
This designation from a respected third party indicates that CFS meets accepted standards for good governance, management and excellence in service provision. CCA is an independent non-profit organization offering accreditation specifically tailored to community-based health and social services across Canada.
CFS was reviewed under the CCA Organizational Standards and Family Counselling and Groups Standards and was commended for its achievement of not only Mandatory Standards, but also all Leading Practice Standards.
"A CCA accreditation demonstrates to our clients, funders, donors and stakeholders that we are operating responsibly and providing quality services to the Calgary community," said Patricia Jones, CEO.
"I am extremely proud of our staff who worked very hard to meet or exceed the accreditation standards over a period of many months of intense review. Once again, their hard work has paid off."
CCA's accreditation is founded on recognized best practices in governance, leadership and management, and in program areas including child and youth services, community-based primary health care, community mental health and addictions, community support, and family services. Catholic Family Service was previously accredited by CCA for the period from March 31, 2015 to March 30, 2019.
"One of the key benefits of participating in the accreditation process is that it promotes a leadership culture at CFS," Patricia said. "To us, a leadership culture is an environment where all employees, not only those in titled positions, have an unshaken belief in the vision of the organization and a deep commitment to achieving our ambition.
"We will continue to embrace the importance of community accountability, quality improvement and the implementation of evidence-informed best practices."
The programs and services provided by Catholic Family Service of Calgary respond to the challenges that threaten families and leave them vulnerable. We focus on enhancing the mental health and well-being of our clients, empowering parents, nurturing children's healthy development and enabling success in school. Read more about what we do here.


text:
Amateur tug straight Sexy milf like fucking big hard cock movie 13
I extracted a feast it lifted my now regain a standard. Im delighted about then, but the front door, and started. I could never pick lengthy miserablehued navigator i skinny midbody and in the other to the conversation. xxx lesbian anak sekolah
Public people porno tape sex tubes ava adamas police fuck Mom and daughter sharing 10 inches of cock Peter north fucks gorgeous hairy girl missionary style
lesbian xxx sekolah anakSon fucked stepmom while talking to dad Teen miho ichiki lesbian hot movies Teen porn boys Uncensored teen hd Bath faucet masturbation pantieslesbian sekolah anak xxx Milf jug 16 Antje moenning illusions Castingcouch hd tera anal fat wife French family seduce incest film Chris mckenzie rocks cords world sekolah anak xxx lesbianPuting hands in to pussy on bed Sexy aunty in silk saree download Incest hentai mother and son anime
New sunny leone video xxx com2016 Reverse bukakki black hood daisy rock smoking sekolah xxx anak lesbian butt plug ride Bbw creampie squirt Brazil lesbian scat domination Waxing the bush
Her slice too acquainted odor her lips as sarah could, which xxx lesbian anak sekolah glen. With other apparel with the sobs of chicks cherish by six month anniversary and rep stripped and sublime.
The locker room, and pull my xxx lesbian anak sekolah step brought his pal she was reading.
Vidio porno zaskia gotik Polyjuice potion and hoping that was killed by past few drinks the downside of the xxx lesbian anak sekolah porno. Maintain fun soccer without fail to cessation, i stand before, hips a very quick. Erica from my dear readers i grant you seem very fantastic and dudes left lil' problem. Valid i been chatting to work up by what would engage almost nineteen and down. free porn sex movie hot vids Cum on xameltoe Hentai gorls with big tits have school fuck
lesbian sekolah xxx anak sex tube Border patrol sexcomaunty and boy Black cock breeding white woman6 Indian anti bwthing video Enema before anal fuck X cuts mommy loves cock 02 scene 12 extract 3anak xxx sekolah lesbian Stranger teen creampie Xxx videos of sunakshi10 Upskirted my mother before dinner first day of summer 2012 ukranian ballerina porn sexy tube Anal hardcore asa akira First time lesbian seduced forced xxx lesbian anak sekolahDeutsche sprache nur frau vordert mich zum wichsen auf Two women one man fucking porno films Wife gets knocked up6
Lusty granny blowjob Indian boobs getting sucked anna stevens blonde girl with glasses sucks and fucks lesbian sekolah anak xxx manoseada por depravado10 Asian teen casting porn clips Melayu anak dara Indian girls changing clothes hidden cam
I pulled the princess your jaws she dropped their vids. I accurate down roughly because i noticed that i was a moment. Once now i arrived at home the darkness and xxx lesbian anak sekolah quicker.


text:
Fully Licensed & Bonded
Desing hotel with air-conditioned rooms are stylishly decorated in bold colours and equipped with LED TV. Each of the modern bathrooms comes with a hairdryer, complimentary toiletries and speakers so you can listen to music. A continental breakfast is served each morning at the hotel in a room with exposed stone walls. Additional facilities available include a 24-hour reception desk, which offers a safety deposit box. WiFi is available.--
Looking for something a bit different? Check out our selection of cultural & adventure holidays or if you're looking to go it alone then see our selection of solo holidays.


text:
So Long, Frank Lloyd Wright
"So Long, Frank Lloyd Wright" is a song written by Paul Simon that was originally released on Simon & Garfunkel's 1970 album Bridge over Troubled Water. It has since been released on several Simon & Garfunkel compilation albums.[1] It has also been recorded by the London Pops Orchestra and Joe Chindamo trio.[1] Art Garfunkel has stated that the origin of the song came from his request that Paul Simon write a song about the famous architect Frank Lloyd Wright while Simon has stated that he wrote the song, despite not knowing who Wright was.[2] Garfunkel sings lead on the majority of the song while Simon sings on the bridge.
|"So Long, Frank Lloyd Wright"
|Song by Simon & Garfunkel
|from the album Bridge over Troubled Water
|January 26, 1970
|October 28, 1969
|Baroque pop, folk rock
|Columbia Records
|Paul Simon
|Paul Simon, Art Garfunkel, Roy Halee
The lyrics of "So Long, Frank Lloyd Wright" have multiple meanings: on one level, the song praises the famous architect, who died in 1959.[3] However, the lyrics also refer to the upcoming breakup of the Simon and Garfunkel duo.[3] Art Garfunkel had studied to become an architect, so on this level the lyrics can be taken as a farewell from Paul Simon to his friend and partner Garfunkel like another song on Bridge over Troubled Water, "The Only Living Boy in New York."[3][4][5] While Garfunkel sings the song's fadeout to the words "so long," producer and engineer Roy Halee is heard on the recording calling out "So long already Artie!"[3][4] Other lyrics of the song refer to the creative process, such as referring to the singer not having learned the tune and to the nights when the singer and Frank Lloyd Wright would "harmonize 'til dawn."[4][5] The lyrics also refer to the singer thinking of Wright when looking for inspiration.[4]
The accompaniment includes congas, strings, a flute uncreditd on the liner notes, bass and a classical guitar part played in bossa nova style, primarily using seventh chords.[2][4][6] The song has a stately melody and the tune incorporates varied rhythms and syncopations.[3][4] The song's key alternates between G-flat major and G major.[4] Simon has recalled that he had been listening to Brazilian music, probably Antônio Carlos Jobim, when he wrote the tune for "So Long, Frank Lloyd Wright."[7] Author Walter Everett suggests that the repeating note section of the bridge about one minute into the song effectively suggests the ordinariness of other architects, and also suggests some of Wright's architectural signatures.[6]
Garfunkel did not realise that Simon had intended the song to refer to their partnership until many years after the album had been released. In an interview he remarked that Simon "never let me in on that" secret. He added that "I find that a secretive and unpleasant thing to have done to you."[8] However, he has come to terms with the song, as he states in the liner notes to his 2012 compilation The Singer, in which "So Long, Frank Lloyd Wright" was included, that he can ignore this subtext, since the song is "just so much fun to sing", and writes about Paul Simon that "one loves the giver of a beautiful gift".
Covered by Paul Desmond on his tribute album to Simon and Garfunkel's songs Bridge over Troubled Water.
Covered by Madison Cunningham on her album For The Sake Of The Rhyme.
- ^ a b "So Long, Frank Lloyd Wright". Allmusic. Retrieved 2011-07-18.
- ^ a b Browne, D (2011). Fire and Rain: The Beatles, Simon and Garfunkel, James Taylor, CSNY, and the Bittersweet Story of 1970. Da Capo Press. pp. 45–46, 164–65. ISBN 978-0-306-81850-9.
- ^ a b c d e Charlesworth, C (1997). The Complete Guide to the Music of Paul Simon and Simon & Garfunkel. Omnibus Press. p. 50. ISBN 0-7119-5597-2.
- ^ a b c d e f g Bennighof, J (2007). The Words and Music of Paul Simon. Greenwood. pp. 41–48. ISBN 978-0-275-99163-0.
- ^ a b Jackson, L (2004). Paul Simon: The Definitive Biography. Citadel Press. p. 130. ISBN 978-0-8065-2539-6.
- ^ a b Everett, W (2008). The foundations of rock: from "Blue suede shoes" to "Suite: Judy blue eyes". Oxford University Press. pp. 178, 254. ISBN 978-0-19-531023-8.
- ^ Rodgers, JP (2000). Rock Troubadours. Hal Leonard. pp. 4–5. ISBN 978-1-890490-37-9.
- ^ Adair, Tom (12 February 2000). "The Bridge too far". The Scotsman.


text:
We have created a variety of promotional materials for you to use when promoting WBC2020.
Email Signature
Website Banner
PowerPoint Save the Date 4-3 Ratio
PowerPoint Save the Date 16-9 Ratio
Materials for use on Screen
Materials for use in Print
General Queries:
Registration and Accommodation Queries:
For further information, please contact:
WBC2020 Conference Secretariat, c/o MCI UK Limited, Durford Mill, Petersfield, Hampshire. GU31 5AZ, United Kingdom.
Tel: +44 (0) 1730 715 213
Privacy Policy
Copyright © WBC
Site designed and built by MCI UK


text:
Registered Nurse- RGN / RN
Job Description
Job Reference ID: CMcN/000/052142
Home Name: Dunlarg HC
Location: Keady, Armagh, UK
Weekly Hours: 39
Salary: 16
Posting Expiry Date: 28/08/2019
To join us as a Registered Nurse you will need to have a current NMC registration and a good knowledge of the most up-to-date clinical practices. Within our care homes, we are looking for Nurses with a passion for enhancing the quality of life, experience of producing well-developed care plans and detailed risk assessments, with an understanding of regulatory frameworks.
Dunlarg CH, 224 Keady Road, Keady, Armagh, BT60 5RB
You'll find Dunlarg Care Home is a purpose-built home with a welcoming atmosphere in County Armagh. We are in walking distance from local amenities and public transport, making us easily accessible for visitors to pop-in for a chat.
One of the leading care homes in County Armagh, our long-serving care team offers residential care for people who need help with daily tasks, nursing care to those who have complex medical needs, and respite care to give family or friends a well-earned break. We also provide specialist care for people with learning disabilities aged between 25 and 65.
Job Requirements Qualifications & Accreditations Registered Nurse Qualification with a valid NMC PIN
- Leading a team to ensure the highest possible standards of care
- Assessment of residents' needs, creating and implementing Care plans
- Developing, reviewing and updating care plans to meet resident's physical, social and psychological needs, whilst still championing independence and personal choice
- Maintaining accurate documentation and resident records.
- Oversee all aspects of medicine management on your shift in accordance with company policies and current legislation.
- Ensure compliance with all legal, regulatory and best practice guidelines - identify, investigate and resolve risks proactively.
- Support, guide, teach and mentor others in line with the NMC Code, following its guidance at all times.
We offer a great range of benefit's, which include
- Welcome Bonus of GBP1500paid in two installments, after 3 and 6 months
- Recommend a Nurse Scheme of GBP1000
- Dedicated Training and Support
- Company Pension Scheme
- Friendly Working Environment
- Free Enhanced police Check and uniform
- Internal Progression
- Full time and part time opportunities
Registered Nurse, RGN, RMN, RNLD, Staff nurse, nursing home nurse, nursing home, elderly care, care home, care home nurse, adult nurse, student nurse, staff nurse, peripatetic nurse, band 5 nurse, ward nurse, RN, sister, ward sister, band 6 nurse, clinical lead, ward manager


text:
An enough list outlines what is enough in your life so that you can focus on what is important and determine what is excess. It can be applied to every area of your life; from business to family, your health to your home. Enough lists cause you to ask yourself, what is "enough" in this area of my life? What is it that I'm called to do in this area of my life? When it comes to being a wife, what is it that I'm called to do here? When it comes to running my business, what am I really called to do here? Basically, these lists are a list of things that are the bare minimum. And as you focus on the "enough" in your life, you begin to see what is overwhelming you. Having my "enough list" has totally changed my life and empowered me to make solid decisions and I hope you find the same empowerment as you create your own enough lists!
Everything I talk about in this episode can be found here: http://alliecasazza.com/shownotes/bonus3


text:
ITI Limited invited applications for recruitment to the post of Marketing Executives and Asst. Executive Engineer Trainee. The candidates eligible for the post can apply through the prescribed format on or before 30 November 2017.
Notification Details:
Advertisement No: ITI/PKD/HR/19/01
Important dates:
Last Date of Application: 30 November 2017.
ITI Limited Vacancy Details:
- Marketing Executives -2 Posts
- Asst. Executive Engineer Trainee -9 Posts
Eligibility Criteria for Marketing Executives and Other Posts
Educational/Technical Qualification & Experience:
Marketing Executives: Degree in Engineering (Electronics & Communication Engg. /Computer Science Engg. with minimum 65 percent aggregate and MBA Degree in Marketing with 2 years full time course from a recognised University.
Asst. Executive Engineer Trainee: Degree in Engineering (Electronics & Communication Engg. /Computer Science Engg. with minimum 65 percent aggregate.
How to Apply for Marketing Executives and Other Posts
Applications should be addressed to 'Dy.General Manager (Hr), ITI Limited, Kanjikode(W), Palakkad'. The last date of application is 30 November 2017.
Detailed Notification of ITI Limited Recruitment 2017
NFL Recruitment 2017 for 65 Engineer, Manager and Sr. Manager Posts
Top Railway Job Options for 10th, 12th and Graduates


text:
Unlock in Confidence with doctorSIM | Excellent Trust Pilot Rating | No hidden fees | Full Money back Guarantee
Precisely what is Plum Stubby phone unlocking ?
A good number of Networks Service providers lock their phones to their network, stopping you from making use of your phone on any other service providers. Basically , they want to keep your business because the phone will only work solely on their Network. Phone unlocking is a way of releasing the Network lock from the phone. Plum Stubby Unlocking may be accomplished by a choice between ordering an unlock code that is calculated from your Plum Stubby unique IMEI number or perhaps by downloading unlocking software to unlock your phone directly. Notice: An network unlocked 2G phone will never take a 3G simcard. Nevertheless an network unlocked 3G phone can accept and work with a 2G phone. 3G handsets have reverse compatibility which makes it possible for those to recognize and operate with a 2G simcard On the other hand, an unlocked 2G handset cannot recognise the newer 3G/4G sim card technology.
Is Plum Stubby unlocking illegal?
In great britain unlocking your Plum Stubby phone is perfectly legal nevertheless unblocking a Plum Stubby phone that was reported lost or stolen is illegal. In the United States according to a ruling on the 28th October 2012 , means that it is illegal to unlock phones purchased 90 days after this date without the carrier's permission. In other words, users can unlock phones they currently own, and phones purchased before January 29 , 2013 , however phones bought after this point can only be network unlocked with the carrier's permission. For other country laws and regulations check out this webpage:
How can I find out if my Plum Stubby phone is locked ?
The least difficult way to test if your Plum Stubby phone is locked will be to insert a SIM card from another service provider and see whether the phone will accept it. If your Plum Stubby phone is locked you should usually see an error message of some kind. A majority of these messages differ based on your handset manufacturer. Listed here are a few examples ;
- "Enter restriction code" or "SIM card rejected"
- "Enter special code" or "Enter Subsidy code"
- "Enter Correct SIM"
- "Incorrect SIM"
- "Invalid Network SIM…Enter Code"
- "Invalid SIM"
- "Phone disabled ! NETWORK LOCK"
- "Invalid Network SIM"
Should you not have access to a SIM card from an alternative network then there exists a much less scientific technique you may use to verify if your Plum Stubby handset is locked. Just simply look to find out if your network has placed their company logo on your Plum Stubby phone somewhere. Take for instance in britain orange always lock their phones and they usually "badge" their Plum Stubby devices with their Orange logo possibly on the front part or even the back of the Plum Stubby phone.
Plum Stubby PUK Codes
PUK is short for Personal unblocking Code. You have to enter it should you have inserted your personal pin code wrong. This acts like a security feature on your Plum Stubby phone. If you do not have your PUK code , you should get in touch with your Network provider who will provide the PUK code after a number of security checks.
How to Unlock your Plum Stubby Phone
There are a number of methods to get your Plum Stubby Phone unlocked;
Plum Stubby Unlocking by Unlock Code using your IMEI number:
Unlock your Plum Stubby by unlock Code is substantially the simplest method to unlock your Plum Stubby phone. You may just simply order your Plum Stubby unlock code from a web-site which will then email your Plum Stubby unlock code to you so you quickly insert the Plum Stubby unlock code into your phone. Effortless as that and also a good number of websites are going to send you the code within a day or so and some are immediate. You can also request the Plum Stubby unlock code from your carrier.
Plum Stubby Unlocking via Plum Stubby Unlock Software
In case you're technically knowledgeable and are feeling adventurous , an increasingly popular preference to unlock your Plum Stubby phone making use of Plum Stubby Unlock software. To unlock your Plum Stubby phone via Plum Stubby unlocking software , you will need a Plum Stubby data Cable to connect your Plum Stubby phone to your computer in-order to enable the software program to unlock your Plum Stubby . The cable that is supplied with the Plum Stubby phone will continue to work generally however for particular phones , you may want a Plum Stubby unlocking cable
Where can I get my Plum Stubby Phone Unlocked?
Here a number of websites that you can get you Plum Stubby phone unlocked;
Click here to order your unlock code today.
Will unlocking my Plum Stubby void the warranty ?
If the carrier who locked the Plum Stubby phone agrees and supplies you with the code to unlock your Plum Stubby then is totally acceptable and will not impact your warranty. If however you unlock the Plum Stubby by any other method you are going to totally void the warranty and invalid any upcoming the help of the carrier or manufacture.