Response shift in oral health-related quality of life measurement in patients with partial edentulism.

The purposes of this study were to determine whether a response shift was observable after partial denture treatment and to identify the predictors that influenced the response shift magnitude and direction. A total of 173 consecutive patients with no more than eight missing teeth who received implant-supported, fixed or removable partial dentures at Okayama University Dental Hospital were asked to complete a full-version Oral Health-Related Quality of Life (OHRQoL) questionnaire before (pre-test) and after treatment (post-test). Additionally, a short form (then-test) consisting of seven questions selected from the full version had its reliability verified and was utilised to retrospectively assess the pre-treatment OHRQoL status. The difference between the summary scores of the then-test and the pre-test determined the response shift magnitude and direction. The then-test mean score (22·9 ± 6·6) was significantly lower (worse OHRQoL) than that of the pre-test (26·4 ± 5·2). The response shift effect size was of moderate magnitude and negative direction (d = -0·78). A multiple regression analysis showed that age (younger patients) (P < 0·01), number of replaced teeth (fewer) (P < 0·01) and pre-test scores (lower) (P < 0·01) were the significant predictors for response shift. In conclusion, a response shift phenomenon with negative and moderate effect size was observed after partial denture treatment. The significant predictor variables were young age, fewer numbers of replaced teeth and lower pre-test scores.

Rapid quantification of polyhydroxyalkanoates (PHA) concentration in activated sludge with the fluorescent dye Nile blue A.

The present study was conducted (1) to develop a rapid quantification method of polyhydroxyalkanoates (PHA) concentration in activated sludge by Nile blue A staining and fluorescence measurement and (2) to perform on-line monitoring of PHA concentrations in activated sludge. Activated sludge samples collected from laboratory scale sequencing batch reactors and full-scale wastewater treatment plants were stained with Nile blue A and their fluorescence intensities were determined. There was a high correlation (R2 > 0.97) between the fluorescence intensities of Nile blue A and PHA concentrations in activated sludge determined by gas chromatography. The Nile blue A staining and fluorescence measurement method allows us to determine PHA concentrations in activated sludge within only five minutes and up to 96 samples can be measured at once by using microplate reader. On-line monitoring of PHA concentrations in activated sludge was achieved by using a fluorometer equipped with a flow cell and the time point at which PHA concentration in activated sludge reached the maximum level could be identified. In addition, we examined the influence of pH, floc size and co-existing chemicals in activated sludge suspension on the fluorescence intensities of Nile blue A.

Acetate uptake by PHA-accumulating and non-PHA-accumulating organisms in activated sludge from an aerobic sequencing batch reactor fed with acetate.

The present study was conducted to evaluate the specific acetate uptake rates of microorganisms with and without polyhydroxyalkanoates (PHA) accumulation. Activated sludge was aerobically incubated with 75 mgC L(-1) radiolabeled or non-labeled acetate, and acetate consumption and PHA accumulation were monitored. Microorganisms were quantified as follows: all microbial cells by DAPI staining, whole acetate utilizing organisms by microautoradiography, and PHA-accumulating organisms by staining with Nile blue A. The abundance of acetate-utilizing organisms without PHA accumulation was also calculated from the outcomes. The estimate of acetate utilized by PHAAOs included both the acetate converted to PHA and that used to supply reducing power and ATP. Acetate utilized by PHAAOs and non-PHAAOs were divided by their respective abundances to obtain their respective specific acetate uptake rates: PHAAOs ranged between 5.3 and 8.0 x 10(-10) mgC cell(-1) h(-1), and non-PHAAOs ranged between 2.8 and 4.2 x 10(-10) mgC cell(-1) h(-1).

Fifteen-year survival of resin-bonded vs full-coverage fixed dental prostheses.

PURPOSE: We clarified cumulative survival and event-free rates of resin-bonded fixed dental prostheses (RBFDPs) and compared them to those of fixed dental prostheses (FDPs) to refine risk factors for non-survival/event and use of tooth extraction after the period of non-survival/event. METHODS: Study subjects were selected among all patients who consecutively attended the Fixed Prosthodontic Clinic of Okayama University Hospital. Eligible patients were those who received 3-unit metal-framed 2-retainer (wing-wing) RBFDPs or conventional full-coverage FDPs (RBFDPs/FDPs: 129/177 prostheses). Data were analyzed by Kaplan-Meier analysis with the log-rank test, Mann-Whitney test, chi-square test, and Cox proportional hazards analysis. RESULTS: The 15-year cumulative survival rates were 66.5% for the RBFDP group and 61.6% for the FDP group, which were not significantly different (p = 0.59). The 15-year cumulative event-free rates were 53.4% for the RBFDP group and 59.2% for the FDP group, which were not significantly different (p = 0.52). No significant risk factors related to non-survival and event-free of RBFDPs/FDPs were identified in the analysis model using treatment method, sex, age, number of remaining teeth, and treatment site as explanatory variables. The number of cases in which RBFDPs/FDPs resulted in non-survival due to abutment tooth extraction was significantly lower in RBFDPs (p < 0.01). Further, the abutment tooth as a non-vital tooth was identified as a risk factor for RBFDPs/FDPs resulting in non-survival due to abutment tooth extraction. CONCLUSIONS: The present study is the first to indicate RBFDP as a prosthetic treatment option which should be selected for patients with slight or no abutment tooth decay.

Rapid quantification of total viral DNA in the supernatants of activated sludge samples with the fluorescent dye PicoGreen.

AIMS: To develop a rapid and simple method for quantifying viral DNA concentrations and determining viral quantities in activated sludge. METHODS AND RESULTS: Activated sludge samples were obtained from three full-scale and one laboratory-scale process. They were centrifuged and the supernatant was filtered through a 0.2-microm membrane filter. Free DNA was removed by DNase-I treatment; any DNA within the viral capsid was liberated by heat treatment and proteinase K, and viral DNA concentrations were determined using the dye PicoGreen. To validate the method, we assessed the recovery of T4 phage added to filtered samples, which was 99% of those added. Viral DNA concentrations in samples from full-scale plants ranged from 69 to 157 ng ml(-1). Monitoring of laboratory-scale reactor samples revealed that viral DNA concentrations varied with time. Our method involves a simple sample treatment protocol and allow rapid analysis of many samples. CONCLUSIONS: A simple, rapid and sensitive method was developed and successfully used to determine the viral DNA concentrations in activated sludge. SIGNIFICANCE AND IMPACT OF THE STUDY: This method provides a way to investigate impact of bacteriophages on the performance of wastewater treatment processes.

Environmental assessment of sewage sludge recycling options and treatment processes in Tokyo.

Tokyo has historically suffered from a shortage of final disposal sites for the treated sewage sludge. Given this situation, sludge recycling and incineration have been promoted to reduce the volume of treated sludge conveyed to the disposal site, and the recycling options have changed since the late 1990s. This study aims to revisit the sewage sludge treatment and recycling processes in Tokyo and to evaluate different recycling options (brick, aggregate, refuse derived fuel and slag) from the energy consumption perspective by clarifying the complex flow of treated sludge within Tokyo's 23 wards. The study also estimates environmental loads associated with the operation of the whole sludge management system in the area. The environmental loads include: (1) total energy consumption and (2) gas emissions (greenhouse and acidification gases). The estimation was carried out for the years 1995, 1997, 1999 and 2001, during which a drastic change in recycling options occurred. The results indicated that the production of refuse derived fuel was the most energy consuming recycling option while aggregate production is the least energy consuming. They also showed that despite the increasing sludge volume, the energy consumption associated with the operation of the whole system decreased during the period while the gas emissions increased.

Sludge-sludge interaction in the enhanced biological phosphorus removal process.

Metabolisms related to enhanced biological phosphorus removal (EBPR) were found to be affected when two activated sludges with different EBPR activities were mixed together. In the present study, two laboratory scale EBPR processes were operated in parallel, one of them with higher and another with lower EBPR activities. The activated sludges from the two reactors were mixed together at different mixing ratios. The supernatant was made the same for all mixing ratios, anaerobic-aerobic batch experiments were performed, and acetate uptake rate and phosphate release rate under anaerobic conditions and phosphate uptake rate under aerobic condition were determined. The metabolic rates measured were expected to be linear to the mixing ratios, as the supernatant was the same for all mixing ratios, whereas the metabolic rates were either promoted or inhibited by mixing of sludges. As an indicator for the sludge mixing effect on the metabolic rates, mixing effect intensity (MEI) was introduced. Chemical substances that are produced by microorganisms in activated sludge are proposed to be one of the possible causes of the sludge mixing effect.

Rain events and their effect on effluent quality studied at a full scale activated sludge treatment plant.

The effect of rain events on effluent quality dynamics was studied at a full scale activated sludge wastewater treatment plant which has a process solution incorporating pre-denitrification in activated sludge with post-nitrification in trickling filters. The incoming wastewater flow varies significantly due to a combined sewer system. Changed flow conditions have an impact on the whole treatment process since the recirculation to the trickling filters is set by the hydraulic limitations of the secondary settlers. Apart from causing different hydraulic conditions in the plant, increased flow due to rain or snow-melting, changes the properties of the incoming wastewater which affects process performance and effluent quality, especially the particle removal efficiency. A comprehensive set of on-line and laboratory data were collected and analysed to assess the impact of rain events on the plant performance.

Examining substrate uptake patterns of Rhodocyclus-related PAO in full-scale EBPR plants by using the MAR-FISH technique.

While recognised as the important population responsible for enhanced biological phosphorus removal (EBPR), detailed knowledge on the physiology of Rhodocyclus-related polyphosphate accumulating organisms (PAO) has yet to be grasped. The objective of this study was to examine the in situ substrate uptake patterns of Rhodocyclus-related PAO present in full-scale EBPR plants by the combined technique of microautoradiography-fluorescent in situ hybridization (MAR-FISH). The presence of these PAO in the four investigated plants was confirmed by FISH and they constituted 17%, 9%, 8%, and 7% of the sludge community. By using MAR-FISH technique, Rhodocyclus-related PAO in all the plants demonstrated similar anaerobic substrate uptake patterns. They were capable of assimilating acetate, aspartate and glutamate under anaerobic condition but they showed negative uptake with palmitate. A significant fraction of the MAR-positive cells assimilated acetate, aspartate or glutamate was found to be Rhodocyclus-related PAO. Dual staining with DAPI and FISH showed that these PAO also accumulated polyphosphate aerobically with aspartate and glutamate as carbon source. The ability of assimilating amino acids besides acetate strongly indicates the versatile physiology of Rhodocyclus-related PAO, which could benefit them to achieve predominance in EBPR activated sludge.

Microbial community of biological phosphorus removal process fed with municipal wastewater under different electron acceptor conditions.

The microbial community in a biological phosphorus removal process under different electron acceptor conditions was estimated by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) assay and principal-component analysis (PCA). For this purpose, a lab-scale sequencing batch reactor (SBR) fed with municipal wastewater was operated under anaerobic-aerobic, anaerobic-anoxic-aerobic and anaerobic-anoxic conditions. The results of PCR-DGGE targeting the 16S rRNA gene indicated a significant shift in the microbial community with electron acceptor conditions. From the 16S rRNA-based PCA, the microbial shift implies that little oxygen supply caused the deterioration of aerobic bacteria, including aerobic polyphosphate-accumulating organisms (PAOs). Moreover, it also reflects the existence of nitrate-utilizing denitrifiers. On the other hand, although the band patterns of DGGE targeting a functional gene of denitrification (nirS) also showed the microbial shift, the result of PCA differed from that of 16S rRNA-based analysis. There is no conclusive proof that the bacteria represented as the dominant bands detected in the present study are denitrifying-PAOs so far, it should be worthwhile to identify the detected bacteria and to examine their traits as new denitrifying-PAO candidates.

Influence of flocculation and settling properties of activated sludge in relation to secondary settler performance.

Floc characteristics were studied at a full scale activated sludge treatment plant with a unique process solution incorporating pre-denitrification with post-nitrification in nitrifying trickling filters. Since greater nitrogen removal is achieved when more secondary settled wastewater is recirculated to the trickling filters, the secondary settlers are always operated close to their maximal capacity. The flocculation and settling properties are therefore crucial and have an effect on the overall plant performance. Since the plant is operated at a short sludge age, these properties change quickly, resulting in variable maximal secondary settler capacity. The dynamics in floc structure and microbial community composition were studied and correlated to the secondary settler performance. Fluorescence in situ hybridisation was used to investigate the microbial community structure and their spatial distribution. The floc structure could to some extent be related to the flocculation and settling properties of the sludge. Even small differences had an influence suggesting that colloidal properties also play a significant role in determining the floc properties. No correlation between microbial community composition and settling properties could be established with the group-specific probes investigated.

Filter clogging in coarse pore filtration activated sludge process under high MLSS concentration.

Coarse pore filtration activated sludge process is a type of hybrid process in which the secondary settling tank of the conventional activated sludge process is replaced by non- woven and coarse pore filter modules. The filter has pores, which are irregular in shape, and much bigger than micro-filtration membrane pores in size. The objective of the study is to find out the effect of the microbial community structure on filter clogging in the coarse pore filtration activated sludge process under high MLSS concentration in aerobic and anoxic/aerobic (A/A) conditions. Filter clogging started from day 65 and 70 in the A/A and aerobic process, respectively, but it was more severe in the A/A process compared to that in the aerobic process. EPS contents of sludge did not change significantly during the operation in both processes, and did not have a crucial effect on the observed filter clogging. There was no strong evidence for direct effect of the type and number of metazoa on filter clogging. The main difference between aerobic sludge and A/A sludge during the filter clogging period was the relative abundance of filamentous bacteria. According to the obtained results, it can be concluded that a higher presence of filamentous bacteria could reduce the severity of filter clogging in a coarse pore filtration activated sludge process.

Resting T cells negatively regulate osteoclast generation from peripheral blood monocytes.

There is accumulating evidence that T cells may be involved in osteoclastogenesis in a variety of murine systems. However, the precise role of human T cells in the regulation of osteoclast generation is still unclear. To address this issue, we investigated the effect of resting peripheral T cells on receptor activator of NF-kappaB ligand (RANKL)-induced osteoclast generation from human peripheral monocytes. Although osteoclasts were not generated in the culture of human peripheral blood mononuclear cells (PBMC) in the presence of RANKL and macrophage colony-stimulating factor (M-CSF), the addition of cyclosporine A (CsA), a potent inhibitor of T-cell function, resulted in the formation of an increasing number of lacunae resorption on dentine, suggesting T cells may inhibit osteoclast formation. In a coculture of T cells and monocytes, which were isolated from PBMC, T cells inhibited the osteoclast generation from monocytes, as determined by tartrate-resistant acid phosphatase (TRAP) staining and a pit assay using dentine. This inhibition of osteoclast generation by T cells was also observed in a culture of the parathyroid hormone-stimulated SaOS4/3 osteoblast cell line and monocytes. The culture in Transwell plates revealed that the cell-to-cell interaction was not required for the inhibition, suggesting that T-cell cytokines may be responsible for the inhibition. Among inhibitory T-cell cytokines on osteoclastogenesis, granulocyte-macrophage colony-stimulating factor (GM-CSF) and interferon-gamma (IFN-gamma) were actively produced by CD4 T cells but not CD8 T cells in the coculture of T cells with monocytes, and the neutralizing antibodies to these cytokines partially rescued the T-cell-induced inhibition of osteoclast formation. Although CsA did not affect RANKL-induced osteoclast generation in the culture of monocytes alone, it completely rescued the T-cell-induced inhibition of osteoclast formation and strongly inhibited the production of GM-CSF and IFN-gamma. Thus, we demonstrate that resting T cells negatively regulate the osteoclast generation via production of GM-CSF and IFN-gamma by CD4 T cells and that CsA stimulates the osteoclast generation through the inhibition of the production of these cytokines. These findings provide new insight into therapeutic strategies for immunosuppression-induced bone loss in transplant and other diseases.

Monokine stimulation of interleukin-11 production by human vascular smooth muscle cells in vitro.

Human vascular smooth muscle cells (VSMC) are a component of blood vessels, and secrete a variety of cytokines in atherosclerotic loci. Interleukin-11 (IL-11), a member of IL-6-like cytokines, is reported to be involved in inflammation and tissue remodeling, both of which are observed in atherosclerosis. However, no information is available as to the production of IL-11 by VSMC. Therefore, the expression of IL-11 in VSMC is investigated. The amounts of IL-11 protein and mRNA were determined by enzyme-linked immunosorbent assay (ELISA) and Northern blot analysis, respectively. The expression of IL-11 in VSMC was also immunohistochemically determined. IL-1 alpha, transforming growth factor-beta (TGF beta) and, to a lesser extent, tumor necrosis factor-alpha (TNF alpha) stimulated the IL-11 production by VSMC, and the stimulatory effects of IL-1 alpha and TGF beta on IL-11 production were dose-dependent. IL-1 alpha and TNF alpha synergistically augmented TGF beta-stimulated IL-11 production by VSMC. Immunohistochemical staining also revealed the expression of IL-11 protein in VSMC. Furthermore, IL-1 alpha, TGF beta, and TNF alpha induced IL-11 gene expression in VSMC. Because IL-6-like cytokines are reported to be cytoprotective, monokine-stimulated IL-11 may have a potent protective role in atherosclerotic lesions.

Role of the endogenous prostaglandin E2 in human lung fibroblast interleukin-11 production.

Interleukin-11 (IL-11) is known to be a member of the interleukin-6 (IL-6)-type cytokine family. IL-11 is likely to be a major determinant of immune regulation in acute and chronic inflammatory lung diseases, although it is not directly linked with specific disease processes. It has already been shown that although unstimulated human lung fibroblasts did not produce significant amounts of IL-11, the addition of interleukin-1 alpha (IL-1 alpha) and/or transforming growth factor-beta (TGF-beta) stimulated fibroblasts dose-dependently to produce IL-11. Northern blot analysis showed that these stimulators also upregulated IL-11 mRNA expression. As it has been previously reported that IL-1 and TGF-beta stimulate prostaglandin E2 (PGE2) release from lung fibroblasts, we investigate here the role of endogenous PGE2 and the direct effects of the two inhibitors of prostaglandin synthesis, indomethacin and dexamethasone, on IL-11 production by human lung fibroblasts. The addition of indomethacin, a cyclo-oxygenase inhibitor, resulted in significant suppression of IL-11 production and mRNA expression in lung fibroblasts. There was no detectable effect of PGE2 alone on IL-11 levels; however, the suppression of IL-11 production by indomethacin was almost completely reversed by addition of PGE2. In contrast, suppression of IL-11 production by indomenthacin was not reversed by addition of thromboxane B2 and carbocyclic thromboxane A2. In addition, dexamethasone completely suppressed IL-11 production and downregulated IL-11 mRNA. These results suggest that endogenous PGE2 acts as an autocrine stimulus for IL-11 production by human lung fibroblasts activated by IL-1 alpha and TGF-beta.

Isolation, characterization and identification of polyhydroxyalkanoate-accumulating bacteria from activated sludge.

Two novel gram-positive bacteria capable of accumulating poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [poly(3HB-co-3HV)] were isolated from an anaerobic-oxic activated sludge system fed with acetate. Strains Lpha5 and Lpha7 are motile cocci, 1-2 microm in diameter, occurring singly or in pairs. These isolates have doubling times ranging from 0.4-1.7 d and can accumulate in high levels of poly(3HB-co-3HV) (up to 44.7% of cell dry weight) when grown on complex media. Furthermore, these two strains exhibited the rapid substrate uptake and accumulation of storage granules as observed in situ. Under aerobic conditions, about 14.4% (cell dry weight) polyhydroxyalkanoate and 82% (carbon dry weight) cellular carbohydrate were produced from acetate and glucose, respectively. Under anaerobic conditions, poly(3HB-co-3HV) and cellular carbohydrate accumulated when glucose but not acetate was fed. The result of analysis of 16S rRNA sequence revealed that both strains belong to the gram-positive high-G + C group, but are significantly different from their closest phylogenetic relatives, Dermatophilus sp. and Terrabacter sp., to warrant classification as a new species.

PHA production by activated sludge.

The production of polyhydroxyalkanoate by anaerobic-aerobic activated sludge was reviewed concentrating on the biochemical mechanisms and on the trials to increase polyhydroxyalkanoate (PHA) content in activated sludge. The anaerobic aerobic activated sludge system selects microorganisms with the capabilities to couple glycolysis, polyphosphate degradation, and PHA accumulation for anaerobic substrate uptake. Some of the PHA-related metabolisms observed there have not been seen in pure cultures so far. Such metabolisms are the formation of PHA containing 3-hydroxy-2-methylvalerate, and '3-hydroxyvalerate fermentation' in which glucose or glycogen is converted to 3-hydroxyvalerate-rich PHA while yielding energy. The PHA content of activated sludge can be increased up to 62% by applying a microaerophilic-aerobic activated sludge process. PHA production by activated sludge is worth investigation.

Measurement of microorganisms with PHA production capability in activated sludge and its implication in activated sludge model no. 3.

In Activated Sludge Model No.3 (ASM3), it is hypothesized that all heterotrophic microorganisms (X(H)) can store substrate. However, in reality, both microorganisms with and without substrate storage capability (X(H/STO) and X(H/S), respectively) could exist. If the ratio of X(H/STO) in activated sludge is influenced by operational and environmental conditions, kSTO (storage rate constant of heterotrophic microorganisms which is defined in ASM3) may not be a universal parameter and can change from case to case. In this study, polyhydroxyalkanoate (PHA) is assumed as the principal storage product, and the ratio of microorganisms with PHA production capability (X(H/PHA)) in various activated sludges was estimated by the dual staining of Nile Blue A (NB) and DAPI. It was shown that the ratio of X(H/PHA) in sludge varied among different municipal and laboratory activated sludges.

Quantitative estimation of the role of denitrifying phosphate accumulating organisms in nutrient removal.

It has been reported that a proportion of polyphosphate-accumulating organisms (PAOs) can denitrify or utilize nitrate as an electron acceptor. The usage of denitrifying-PAO (DN-PAO) can relieve the competition for COD between denitrification and phosphorus removal because they can treat nitrate and phosphate by using the same carbon source. To effectively use DN-PAO for biological nutrient removal (BNR), a new system was proposed in which an anaerobic phase is placed at the influent end, followed by he anoxic and external nitrification phase. In this study, the lab-scale proposed system (A2N system) was operated with a municipal wastewater 1) to confirm stable settlement of DN-PAO in the proposed system, 2) to quantitatively estimate the fraction of different groups of organisms like denitrifiers without polyphosphate accumulating capability, aerobic-PAO and DN-PAO and 3) to estimate the advantages of DN-PAO's presence in the system. Moreover, batch experiments in which anoxic and aerobic phosphate uptake rates (PUR) were measured were also carried out. The activity of DN-PAO was observed throughout the experimental period by the batch experiment. From the results of the calculation of COD utilized by each group of organisms, it was concluded that the proposed system could accumulate much more PAO (as DN-PAO) than conventional BNR systems. Moreover, they were responsible for both EBPR and denitrification.

Analysis of microbial community that performs enhanced biological phosphorus removal in activated sludge fed with acetate.

Enhanced biological phosphorus removal (EBPR) activated sludge was operated in a laboratory-scale sequencing batch reactor (SBR) fed with acetate as the sole carbon source. The microbial community of the sludge was analyzed using the polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis (DGGE) method for about 2 months of start-up period. As a result, the number of major bands decreased during the enrichment, indicating that the microbial community structure was getting simpler. Since the phosphate removal activity was maintained at a high level, the bacteria which still remained at the end can be considered as the important bacteria playing key roles in the present EBPR sludge, maybe polyphosphate accumulating organisms (PAOs). The dominant band in the last sample on the DGGE gel was excised and the DNA recovered from it was sequenced. The sequence was closely related to one of the putative PAOs group which Crocetti et al. (2000) and Hesselmann et al. (1999) have proposed. This PAOs group is closely related to the Rhodocyclus group (b-Proteobacteria). The fluorescence in situ hybridization (FISH) method with the probe specific for this PAOs group and the DAPI staining at a phosphate-probing concentration indirectly showed that these Rhodocyclus related bacteria really accumulated polyphosphate.