Laundry wastewater and domestic sewage pilot-scale anaerobic treatment: Microbial community resilience regarding sulfide production.

In this study, the removal of anionic surfactant Linear Alkylbenzene Sulfonate (LAS) from laundry wastewater was evaluated in co-digestion with domestic sewage, using a pilot-scale Expanded Granular Sludge Bed reactor. Surfactant influent concentration was enhanced from 5 ±â€¯3 mg LAS L-1 (stage I) to 19 ±â€¯10 mg LAS L-1 (stage II) and 36 ±â€¯19 mg LAS L-1 (stage III) throughout reactor operation. Sulfide levels higher than 20 mg L-1 influenced LAS removal efficiency, which decreased from 71% to 55% and 32% in stage I, II and III, respectively. Acclimation of microbial population was verified and higher relative abundance of the genera similar to Cytophaga, Bacteroides, Syntrophus and Syntrophobacter in the early stages (adaptation and stage I) was replaced by higher relative abundance of the genera Anaerophaga, Nitrosovibrio, Sulfurovum and Desulfovibrio in the last stages (stage II and III).

Metagenomic analysis of the microbiome in three different bioreactor configurations applied to commercial laundry wastewater treatment.

The taxonomic and functional diversity of three different biological reactors (fluidized bed reactor, FBR; up-flow anaerobic sludge blanket reactor, UASB; and expanded granular sludge bed reactor, EGSB) used for commercial laundry wastewater treatment was investigated using metagenome shotgun sequencing. Metagenomes were sequenced on the Illumina Hiseq platform and were analyzed using MG-RAST, STAMP and PAST software. The EGSB and UASB reactors were more closely related based on taxonomic and functional profiles, likely due to similar granular sludge and procedures adopted to ensure anaerobic conditions. The EGSB and UASB reactors showed a predominance of methanogens and genes related to methanogenesis, with a prevalence of the acetoclastic pathway, in addition to the peripheral and central O2-independent pathways for aromatic compound degradation. By contrast, FBR showed a dominance of aerobic microbiota and pathways for O2-dependent aromatic compound degradation. Therefore, although the reactors showed similar surfactant removal levels, the microbial composition, functional diversity and aromatic compound degradation pathways were significantly distinct.

Evaluation of anionic surfactant removal in anaerobic reactor with Fe(III) supplementation.

The objective of this study was to evaluate the removal of linear alkylbenzene sulfonate (LAS) associated with Fe(III) supplementation using an expanded granular sludge bed (EGSB) reactor. The reactor was inoculated with a granular sludge and fed with synthetic wastewater containing a specific LAS load rate (SLLR) of 1.5 mg gVS-1 d-1 (∼16.4 mgLAS L-1 influent) and supplied with 7276 µMol L-1 of Fe(III). The biomasses from the inoculum and at the end of the EGSB-Fe operation (127 days) were characterized using 16S rRNA Ion Tag sequencing. An increase of 20% in the removal efficiency was observed compared to reactors without Fe(III) supplementation that was reported in the literature, and the LAS removal was approximately 84%. The Fe(III) reduction was dissimilatory (the total iron concentration in the influent and effluent were similar) and reached approximately 64%. The higher Fe(III) reduction and LAS removal were corroborated by the enrichment of genera, such as Shewanella (only EGSB-Fe - 0.5%) and Geobacter (1% - inoculum; 18% - EGSB-Fe). Furthermore, the enrichment of genera that degrade LAS and/or aromatic compounds (3.8% - inoculum; 29.6% - EGSB-Fe of relative abundance) was observed for a total of 20 different genera.

The comparative advantages of ethanol and sucrose as co-substrates in the degradation of an anionic surfactant: microbial community selection.

The efficiency of linear alkylbenzene sulfonate (LAS) removal from laundry wastewater and the related microbial community was investigated in an anaerobic fluidized bed reactor (AFBR). The AFBR was operated in three stages, in addition to the biomass adaptation stage without LAS (stage I). The stages were differentiated by their supplementary co-substrates: stage II had sucrose plus ethanol, stage III had only ethanol, and stage IV had no co-substrate. The replacement of sucrose plus ethanol with ethanol only for the substrate composition favored the efficiency of LAS removal, which remained high after the co-substrate was removed (stage II: 52 %; stage III: 73 %; stage IV: 77 %). A transition in the microbial community from Comamonadaceae to Rhodocyclaceae in conjunction with the co-substrate variation was observed using ion sequencing analysis. The microbial community that developed in response to an ethanol-only co-substrate improved LAS degradation more than the community that developed in response to a mixture of sucrose and ethanol, suggesting that ethanol is a better option for enriching an LAS-degrading microbial community.

Microbial diversity and the implications of sulfide levels in an anaerobic reactor used to remove an anionic surfactant from laundry wastewater.

The objective of this study was to evaluate the removal of linear alkylbenzene sulfonate (LAS) from commercial laundry wastewater using an expanded granular sludge bed (EGSB) reactor with two specific LAS loading rates (SLLRs), 1.0 and 2.7 mg LAS gVS(-1)d (-1). The biomass was characterized using denaturing gradient gel electrophoresis (DGGE) and 16S Ion Tag sequencing. Higher LAS removal (92.9%) was observed in association with an SLLR of 1.0 mg LAS gVS(-1) d(-1) than with an SLLR of 2.7 mg LAS gVS(-1) d(-1) (58.6%). A relationship between the S(-2) concentration in the effluent and the surfactant removal efficiency was observed. This result is indicative of the inhibition of LAS-removing microbiota at S(-2) concentrations greater than 20 mg SL(-1). By using DGGE, microbial stratification was observed in the reactor in association with granule size, even though the reactor is considered to be a completely mixed regime. The RDP-classifier identified 175 genera, 33 of which were related to LAS degradation.

Analysis of a microbial community associated with polychlorinated biphenyl degradation in anaerobic batch reactors.

The degradation of polychlorinated biphenyls (PCBs) was investigated under fermentative-methanogenic conditions for up to 60 days in the presence of anaerobic biomass from a full-scale UASB reactor. The low methane yields in the PCBs-spiked batch reactors suggested that the biomass had an inhibitory effect on the methanogenic community. Reactors containing PCBs and co-substrates (ethanol/sodium formate) exhibited substantial PCB reductions from 0.7 to 0.2 mg mL(-1). For the Bacteria domain, the PCBs-spiked reactors were grouped with the PCB-free reactors with a similarity of 55 %, which suggested the selection of a specific population in the presence of PCBs. Three genera of bacteria were found exclusively in the PCB-spiked reactors and were identified using pyrosequencing analysis, Sedimentibacter, Tissierela and Fusibacter. Interestingly, the Sedimentibacter, which was previously correlated with the reductive dechlorination of PCBs, had the highest relative abundance in the RCS-PCB (7.4 %) and RCS-PCB-PF (12.4 %) reactors. Thus, the anaerobic sludge from the UASB reactor contains bacteria from the Firmicutes phylum that are capable of degrading PCBs.

Effect of biomass adaptation to the degradation of anionic surfactants in laundry wastewater using EGSB reactors.

Two expanded granular sludge bed reactors were operated. RAB (adapted biomass) was operated in two stages: Stage I, with standard LAS (13.2 mg L(-1)); and Stage II, in which the standard LAS was replaced by diluted laundry wastewater according to the LAS concentration (11.2 mg L(-1)). RNAB (not adapted biomass) had a single stage, using direct wastewater (11.5 mg L(-1)). Thus, the strategy of biomass adaptation did not lead to an increase of surfactant removal in wastewater (RAB-Stage II: 77%; RNAB-Stage I: 78%). By means of denaturing gradient gel electrophoresis, an 80% similarity was verified in the phases with laundry wastewater (sludge bed) despite the different reactor starting strategies. By pyrosequencing, many reads were related to genera of degraders of aromatic compounds and sulfate reducers (Syntrophorhabdus and Desulfobulbus). The insignificant difference in LAS removal between the two strategies was most likely due to the great microbial richness of the inoculum.

The relationship between bite force and oral sensation during biting in molars.

BACKGROUND: In prosthodontic treatment, the occlusal form should be designed such that bite force is applied at the position that has higher load-bearing capacity and is comfortable for the patient. The purpose of this study was to clarify the differences in bite force and occlusal sensation with different loading positions on the occlusal surface. METHODS: Twelve healthy subjects were recruited for this study. Bite force and occlusal sensation were measured at five loading points on the upper and lower left first molars. Occlusal sensation was evaluated using a Visual Analogue Scale (VAS). RESULTS: Bite forces on the lingual side of the upper first molar and the buccal side of the lower first molar were significantly higher and VAS scores were significantly lower, i.e. the subjects felt less discomfort during biting on the buccal side of the upper first molar and the lingual side of the lower first molar compared to the other side of each molar. CONCLUSIONS: Loading on the occlusal surfaces of the functional cusps of the upper and lower first molars produces more load-bearing capacity and is more comfortable than loading on the non-functional cusps.

Microbial characterization and removal of anionic surfactant in an expanded granular sludge bed reactor.

This study evaluated linear alkylbenzene sulfonate removal in an expanded granular sludge bed reactor with hydraulic retention times of 26 h and 32 h. Sludge bed and separator phase biomass were phylogenetically characterized (sequencing 16S rRNA) and quantified (most probable number) to determine the total anaerobic bacteria and methanogenic Archaea. The reactor was fed with a mineral medium supplemented with 14 mg l(-1)LAS, ethanol and methanol. The stage I-32 h consisted of biomass adaptation (without LAS influent) until reactor stability was achieved (COD removal >97%). In stage II-32 h, LAS removal was 74% due to factors such as dilution, degradation and adsorption. Higher HRT values increased the LAS removal (stage III: 26 h - 48% and stage IV: 32 h - 64%), probably due to increased contact time between the biomass and LAS. The clone libraries were different between samples from the sludge bed (Synergitetes and Proteobacteria) and the separator phase (Firmicutes and Proteobacteria) biomass.

Effect of direct retainer and major connector designs on RPD and abutment tooth movement dynamics.

Designs of removable partial dentures are suggested to affect the mobility of abutment teeth and removable partial denture (RPD) during oral functions. This study aimed to examine the effect of direct retainer and major connector designs on RPD dynamics under simulated loading. Six different Kennedy class II maxillary RPDs were fabricated on a maxillary model. These dentures involved 3 different direct retainers (wrought-wire clasp, RPA clasp, and conical crown telescopic retainer) and 2 different major connectors (Co-Cr major connector and heat-cured acrylic resin with a metal strengthener). Using an experimental model with simulated periodontal ligaments and mucosa that were fabricated using silicone impression material, three-dimensional displacements of the RPDs were measured under a simulated 30-N loading with a displacement transducer type M-3. Significant effects of "direct retainer design" on bucco-palatal displacements and "major connector" on mesio-distal displacements were revealed by 2 x 3 two-way analysis of variance of abutment teeth movements (P < 0.001 and P = 0.002, respectively). Additionally, analysis of variance of RPD displacements revealed significant effects of "direct retainer design" on corono-apical displacements and "major connector" on mesio-distal displacements (P = 0.001 and P = 0.028, respectively). Rigid direct retainers and rigid major connectors decrease the movements of both abutment tooth and RPD.

[Concomitant off-pump coronary artery bypass and left lower lobectomy for lung cancer with pneumoconiosis].

Surgical strategy for patients for lung cancer with coronary disease remains controversial. We performed concomitant off-pump coronary artery bypass grafting (off-pump CABG: OPCAB) and left lower lobectomy with ND 2a lymph nodes dissection for lung cancer with pneumoconiosis. After CABG, due to fixed adhesion of peribronchial lymph node, left lower lobectomy was hard to carry out. Postoperative management was done with much difficulty for massive sputum and pneumonia caused by chronic inflammatory lung disease. As a result, postoperative course has been uneventful for 2 years after operation. In the concomitant OPCAB and lung resection with chronic inflammatory lung disease, whether the synchronous or 2 periods operation was appropriate was still unknown. We think that careful pre- and intraoperative assessment for this disease is important.

Distinct glutamate receptors govern differential levels of nitric oxide production in a layer-specific manner in the rat cerebellar cortex.

To evaluate roles of nitric oxide (NO) in neural functions, it is critical to know how neural inputs activate neuronal NO synthase in individual sites. Although NMDA receptor-dependent mechanism well explains postsynaptic, robust NO production, this sole mechanism does not explain some aspects of NO production in the brain, such as the low-level production of NO and the mechanism for presynaptic NO production. We hypothesized that the glutamate receptor involved in NO production is site-specific and controls the initial NO concentration in each site. We visualized NO production mediated by NMDA, AMPA and type-1 metabotropic glutamate (mGlu-1) receptors in rat cerebellar slices and granule cells in culture, with an NO-specific fluorescent indicator, diaminofluorescein-2. AMPA receptor, but not NMDA or mGlu-1 receptor, was responsible for NO production at parallel fiber terminals, which was blocked by CNQX, tetrodotoxin or voltage-dependent calcium channel blockers. More numbers of electrical stimulation were required for NO production in the molecular layer than in other layers, suggesting that AMPA receptor activation generates NO at lower concentrations through a remote interaction with NO synthase. Although Purkinje cell does not express NO synthase, we detected NO production in Purkinje cell layer following electrical stimulation in the white matter at 50 Hz, but not at 10 Hz. This NO production was tetrodotoxin-sensitive, suggesting occurrence in the basket cell terminals, and required synergistic activation of mGlu-1 and NMDA receptors. In the granule cell layer, activation of AMPA or mGlu-1 receptor produced NO uniformly, while NMDA receptor activation produced NO in discontinuous areas of this layer. Thus, distinct glutamate receptors, including non-NMDA receptors, govern occurrence and level of NO production in a layer-specific manner.

[The evaluation of the complications and its management in 800 patients who underwent the thoracoscopic surgery].

Thoracoscopy has been applied to almost all thoracic surgery as shown (table 1). Besides an opportunity to follow and encounter a trouble during surgery using thoracoscopy, so that it is necessary to become skilled in the prediction and management of complication. Nowadays, there are two options on surgery using thoracoscopy, thoracoscopic surgery (TS) is performed only on in a port and on a mini-thoracotomy with a few ports (video-assisted thoracoscopic surgery: VATS). When changing into emergency or converting into open thoracotomy, a muscle sparing thoracotomy could be available. The symptoms which occurred in the thorax are fundamentally similar during both TS and VATS procedure. However, peculiar dangerous factor existed according to the difference in internal organs, procedures, and diseases. Critical complications during surgery were as follows, i.e., injury of pulmonary vessels and bronchus, tension pneumothorax of the opposite lung and tumor implantation of the chest wall. Basically, standard practice is the most important to avoid an accident and determine whether converting to open thoracotomy should be employed. In the present commentary, we would like to present our experiences and propose regarding management.

Myocardial kinetics of Tc-99m glucarate in low flow, hypoxia, and aglycemia.

BACKGROUND: Technetium-99m glucarate is a myocardial infarct-avid imaging agent. Recent conflicting and inconclusive reports have suggested that the agent may be taken up by ischemic but viable myocardium. The purposes of this study were (1) to determine conclusively whether there is Tc-99m glucarate uptake in ischemic viable myocardium and (2) to investigate the potential mechanisms for such uptake by studying components of ischemia, namely, low flow, hypoxia, and aglycemia. METHODS AND RESULTS: Rat hearts were isolated and perfused in a modified Langendorff preparation with a crytalloid perfusate. Tc-99m glucarate was studied in control (n = 6), low-flow (n = 5), hypoxic (n = 5), and aglycemic (n = 5) conditions. The experimental protocol consisted of 20-minute baseline (12 mL/min flow) and 20-minute treatment (low flow at 1 mL/min, hypoxia, or aglycemia), followed by tracer uptake (20 minute) and washout (20 minutes). Activity was monitored with a sodium iodide detector. The tracer was delivered continuously over a 20-minute uptake period. The injected dose was 150 micro Ci (5.6 MBq). Hemodynamics were monitored throughout. Triphenyltetrazolium chloride staining was used to assess myocardial viability. There was no evidence of myocardial necrosis. Low flow tended to delay tracer uptake compared with control for the first 10 minutes, but this did not reach statistical significance. Low flow increased end fractional retention significantly compared with control (mean +/- SEM, 59.0% +/- 0.9% peak vs 41.2% +/- 1.4%, respectively; P <.05). Hypoxia resulted in a trend toward increased uptake; however, this was significant only at one early time point during the uptake phase. Retention in the hypoxia group was similar to control. Tc-99m glucarate uptake was significantly increased in aglycemia from 16 minutes to peak compared with control (1.36% +/- 0.71% injected dose per gram vs 0.91% +/- 0.37% injected dose per gram, respectively; P <.05). Aglycemia produced significantly higher end fractional retention compared with control (51.6% +/- 1.8% peak vs 41.2% +/- 1.4%, respectively; P <.05). CONCLUSIONS: Tc-99m glucarate myocardial retention is increased in the setting of ischemia, even in the absence of necrosis. This increased retention is not due to hypoxia. Furthermore, the retention is only partially explained by tissue hypoglycemia. Thus low flow per se appears to have a role in this increased retention, probably as a result of delayed flow-dependent washout.

Sast124, a novel splice variant of syntrophin-associated serine/threonine kinase (SAST), is specifically localized in the restricted brain regions.

Syntrophin is an adaptor protein that binds signaling molecules to the dystrophin-associated protein complex, which connects extracellular matrix to intracellular cytoskeleton for construction and maintenance of the postsynaptic structures in the neuromuscular junction and the CNS. Among these signaling molecules, a family of microtubule-associated serine/threonine kinases has a unique structural feature with a serine/threonine kinase domain and a postsynaptic density protein-95/discs large/zona occludens-1 domain. In the present study, we identified syntrophin-associated serine/threonine kinase-124, a novel splice variant of the syntrophin-associated serine/threonine kinase which is a member of the microtubule-associated serine/threonine kinases family. Comparing to the original clone (syntrophin-associated serine/threonine kinase-170), syntrophin-associated serine/threonine kinase-124 is truncated just downstream of the postsynaptic density protein-95/discs large/zona occludens-1 domain. Using a monoclonal antibody specifically recognizing syntrophin-associated serine/threonine kinase-124, strong expression of the protein was observed in neurons of the subventricular zone and granule cells of the olfactory bulb, Islands of Calleja, hippocampal dentate gyrus and cerebellum. syntrophin-associated serine/threonine kinase-124 is selectively localized in the nuclei of neurons and distinct from syntrophin-associated serine/threonine kinase-170, which is interacting with syntrophin on the cell surface. Considering the tissue and subcellular distributions of syntrophin-associated serine/threonine kinase-124, it is suggested that syntrophin-associated serine/threonine kinase-124 may have functions in transcriptional regulation for the features commonly shared by these neurons. On the other hand, syntrophin-associated serine/threonine kinase-124 was also localized in glia-like cell bodies in the corpus callosum and fiber bundles in the spinal trigeminal and solitary tracts, suggesting syntrophin-associated serine/threonine kinase-124 may have other functions in these types of cells.

High survival rate of 6 cases of pulmonary large cell neuroendocrine carcinoma formerly classified as small cell carcinoma.

UNLABELLED: In the revised WHO classification of lung cancer, published in 1999, large cell neuroendocrine carcinoma (LCNEC) was employed as a new histological entity. LCNEC is generally considered a high-grade malignant lung cancer, and appropriate treatment remains to be determined. Before its new classification, LCNEC had long been classified into several entities. Advancing the review of previous cases in Nippon Medical School Hospital, we noticed that some LCNEC patients were formally diagnosed as having small cell lung cancer (SCLC), and they showed long-term survival. MATERIAL AND METHODS: All histological specimens of surgically resected SCLC in Nippon Medical School Hospital were reclassified according to the 1999 WHO classification manual. Their neuroendocrine differentiations were confirmed by the use of immunostainings with chromogranin A and synaptophysin. RESULTS: Fourteen cases satisfied the qualifications for both histological and clinical reevaluation. Among them, 6 patients were reclassified as LCNEC, and their stage distribution was as follow: IA; 1, IB; 2, IIIA; 2, and IIIB; 1. Their survival term ranged from 33.8 to 78.0 months; 5 were still alive, and 1 (IIIB) died 57.6 months after surgery. DISCUSSION: According to this study, all the LCNEC patients who were treated as SCLC patients showed more favorable prognoses than patients described in published studies, even overall lung cancer. Therefore, it is suggested that multimodality therapy for SCLC may improve the prognoses of patients with LCNEC.

Intercellular action of nitric oxide increases cGMP in cerebellar Purkinje cells.

cGMP is thought to play a role in cerebellar signalling yet its production within Purkinje cells has never been detected. In the present study, the hydrolysis of a fluorescent substrate analogue, 2'-O-anthranyloyl cyclic GMP, by type 5 phosphodiesterase was monitored within Purkinje cells in slices and in culture. Nitric oxide, either endogenously released from adjacent neurons or pharmacologically applied, accelerated the rate of hydrolysis in a manner that was dependent on soluble guanylyl cyclase, demonstrating that nitric oxide triggers cyclic GMP production in Purkinje cells, which in turn activates type 5 phosphodiesterase. We conclude that NO acts as an intercellular messenger in the cerebellar cortex and that parallel fibre terminals are a probable source of nitric oxide.

Neutral red as a hydrophobic probe for monitoring neuronal activity.

Neutral red fluorescence has been used to monitor neuronal activity. Local changes in either pH or hydrophobicity are reported to increase neutral red fluorescence, but the mechanisms underlying the increases remain unclear. In this study, the pH-dependent fluorescence changes in the basic and acidic forms of neutral red preloaded in rat cerebellar slices were separately measured with two excitation wavelengths. Bath application of kainate, domoate or alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) at 1 mM increased fluorescence of the acidic form and decreased that of the basic form irreversibly, consistent with a pH-dependent mechanism. In contrast, application of lower concentrations of AMPA, (S)-3,5-dihydroxyphenylglycine or KCl increased fluorescence of both forms transiently, suggesting that a pH-independent mechanism may also increase neutral red fluorescence. Electrical stimulation in the molecular layer also increased fluorescence of both forms. The response to weak electrical stimulation decayed in about 1 s, while response to intense stimulation lasted longer than 1 min. Neutral red binding to phospholipids was also detected as fluorescent spots on thin-layer chromatography, suggesting that interaction with phospholipids enhances neutral red fluorescence. Thus, the short-lasting signal of neutral red suggests its usage as a hydrophobic probe for neuronal activity.