Case study of the relationship between fungi and bacteria associated with high-molecular-weight polycyclic aromatic hydrocarbon degradation

Although bacteria play dominant roles in microbial bioremediation, few of them have been reported that were capable of utilizing high-molecular-weight (HMW) organic pollutants as their sole sources of carbon and energy. However, many soil fungi can metabolize those of pollutants, although they rarely complete mineralization. In this paper, we investigated the dynamic relationship between fungi and bacteria associated with degradation of HMW-polycyclic aromatic hydrocarbons (PAHs). Artificial fungal-bacterial mixed cultures were constructed to simulate the environment of actual polluted sites. Four bacterial strains and seven fungal strains were isolated that related to the removal of phenanthrene, fluoranthene and pyrene in the soil. Furthermore, these strains were used to create mixed culture of bacteria (Bact-mix), mixed culture of fungi (Fung-mix), fungal-bacterial co-cultures (Fung-Bact), respectively. The maximal pyrene removal rate (67%, 28days) was observed in the Fung-Bact, compared with cultures of Fung-mix (39%) and Bact-mix (56%). The same tendency was also indicated in the degradation of phenanthrene and fluoranthene. In addition, a dynamic relationship during the degradation process between fungi and bacteria was monitored through using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) method.     

Construction of hydrogen fermentation from garbage slurry using the membrane free bioelectrochemical system

The aim of this study was to show the effectiveness of the membrane free bioelectrochemical system (BES) using three electrodes on inhibition of methanogenesis and construction of hydrogen fermentation from the artificial garbage slurry. The electrical redox-potential on the working electrode was adjusted to -1.0V (vs. Ag/AgCl) that has positive effect on methanogenesis. The redox-potential on the counter electrode was measured to be 1.6V. The pH in the effluents was 5.5-6.4. Hydrogen production rate at the cathode side was similar to that at the anode side and much higher than that calculated from current, and reached a maximum of 2445±815 (average±standard deviation) mL?L(-1)?d(-1) at an organic loading rate of 58.7g dichromate chemical oxygen demand per L d(-1). Methane production was negligible throughout the experiment. Acetate and butyrate were the main products of the fermentation using a BES; these offered favorable conditions for hydrogen production. The bacterial community in the bioelectrochemical hydrogen fermentor differed from that in the methanogenic seed sludge and included hitherto unknown species. These results show that high redox-potential on the anodic electrode and acidic pH in the membrane free BES can be utilized for hydrogen fermentation from the artificial garbage slurry by avoiding methanogenesis.     

Acceleration of cellulose degradation and shift of product via methanogenic co-culture of a cellulolytic bacterium with a hydrogenotrophic methanogen

Although the effects of syntrophic relationships between bacteria and methanogens have been reported in some environments, those on cellulose decomposition using cellulolytic bacteria from methanogenic reactors have not yet been examined. The effects of syntrophic co-culture on the decomposition of a cellulosic material were investigated in a co-culture of Clostridium clariflavum strain CL-1 and the hydrogenotrophic methanogen Methanothermobacter thermautotrophicus strain ΔH and a single-culture of strain CL-1 under thermophilic conditions. In this study, strain CL-1 was newly isolated as a cellulolytic bacterium from a thermophilic methanogenic reactor used for degrading garbage slurry. The degradation efficiency and cell density of strain CL-1 were 2.9- and 2.7-fold higher in the co-culture than in the single-culture after 60?h of incubation, respectively. Acetate, lactate and ethanol were the primary products in both cultures, and the concentration of propionate was low. The content of acetate to total organic acids plus ethanol was 59.3% in the co-culture. However, the ratio decreased to 24.9% in the single-culture, although acetate was the primary product. Therefore, hydrogen scavenging by the hydrogenotrophic methanogen strain ΔH could shift the metabolic pathway to the acetate production pathway in the co-culture. Increases in the cell density and the consequent acceleration of cellulose degradation in the co-culture would be caused by increases in adenosine 5'-triphosphate (ATP) levels, as the acetate production pathway includes ATP generation. Syntrophic cellulose decomposition by the cellulolytic bacteria and hydrogenotrophic methanogens would be the dominant reaction in the thermophilic methanogenic reactor degrading cellulosic materials.     

Adhesion Properties of Acrylic Copolymers Modified with Si-Containing Copolymer

We made clear the cause of the increase in peel strength of pressure sensitive (PS) adhesives as a function of contact time, and investigated how to modify PS adhesives to maintain a low and constant peel strength for a long time. It was found that polar groups in the adhesive orient to the interface between the adhesive and the (stainless steel) metal substrate (SUS 304) so as to minimize interfacial free energy during adhesion, and the orientation increased affinity between the adhesive and the metal material and increased the peel strength as a result. The use of modifier which contained both P(MMA-co-SiMA) and PDMS showed an excellent modification effect, although modification with only PDMS or P(MMA-co-SiMA) was not sufficient. It was suggested that PDMS which migrated to the surface was extended uniformly over the surface by PDMS segments of P(MMA-co-SiMA) and that the enriched layer of PDMS on the adhesive surface worked as a barrier to prevent the orientation of polar groups in bulk. Therefore, low and constant peel strength could be achieved.     

Design Considerations and Performance Evaluation of a Compact Aerosol Particle Mass Analyzer

A compact aerosol particle mass analyzer (APM) of which the size of the classifier was significantly reduced than that of the first commercial model (Kanomax Model 3600) was developed. Firstly, requirements for desired performance in classifying particle mass were set forth. Secondly, a theoretical framework for the design parameters of an APM that satisfies the requirements was formulated. Thirdly, the design parameters were determined that satisfies the requirements while reducing the instrument size. The requirements include the condition that the classification range covers from 0.001 to 1000 fg (approximately 12 to 1200 nm in size for spherical particles having the density of 1 g/cm³), and the condition that both the classification resolution and particle penetration in this mass range are higher than certain specified values. A prototype having the design parameters determined according to this theoretical framework was constructed, and its performance was evaluated experimentally. The external dimensions of the electrodes of the compact APM are approximately 140 mm in length and 60 mm in diameter. It was confirmed that the performance of the compact APM operated at the aerosol flow rate of 0.3 L/min was comparable to that of the Model 3600 APM operated at 1 L/min. Because of the reduced size and of the resultant improved portability, it is expected that the compact APM is readily applicable to field measurements.

Copyright 2013 American Association for Aerosol Research     

A simple screening procedure for heterotrophic nitrifying bacteria with oxygen-tolerant denitrification activity

Various naturally occurring strains of heterotrophic nitrifying bacteria were isolated by enrichment culture using acetamide as the C and N source, and 21 strains were identified as heterotrophic nitrifiers. Using a new simple procedure, these 21 strains were also investigated for the ability to carry out denitrifcation in the presence of oxygen. Several of the nitrifying strains were found to exhibit a distinct activity that allows for denitrifcation via nitrite (NO2-) in the presence of oxygen, indicating that they have an oxygen-tolerant denitrifcation system. A wide variety of bacteria possessing both nitrification and denitrifcation capabilities in the presence of oxygen were isolated and partially characterized by using the simple screening combinatorial procedure described in this paper.     

Downregulation of miR-122-5p Activates Glycolysis via PKM2 in Kupffer Cells of Rat and Mouse Models of Non-Alcoholic Steatohepatitis

Non-alcoholic steatohepatitis (NASH) has pathological characteristics similar to those of alcoholic hepatitis, despite the absence of a drinking history. The greatest threat associated with NASH is its progression to cirrhosis and hepatocellular carcinoma. The pathophysiology of NASH is not fully understood to date. In this study, we investigated the pathophysiology of NASH from the perspective of glycolysis and the Warburg effect, with a particular focus on microRNA regulation in liver-specific macrophages, also known as Kupffer cells. We established NASH rat and mouse models and evaluated various parameters including the liver-to-body weight ratio, blood indexes, and histopathology. A quantitative phosphoproteomic analysis of the NASH rat model livers revealed the activation of glycolysis. Western blotting and immunohistochemistry results indicated that the expression of pyruvate kinase muscle 2 (PKM2), a rate-limiting enzyme of glycolysis, was upregulated in the liver tissues of both NASH models. Moreover, increases in PKM2 and p-PKM2 were observed in the early phase of NASH. These observations were partially induced by the downregulation of microRNA122-5p (miR-122-5p) and occurred particularly in the Kupffer cells. Our results suggest that the activation of glycolysis in Kupffer cells during NASH was partially induced by the upregulation of PKM2 via miR-122-5p suppression.     

Changes in lipids in liver and serum of rats fed a histidine-excess diet or cholesterol-supplemented diets

The influence of dietary excess (5%) L-histidine on serum and liver lipids was examined in rats. Feeding a histidine-excess diet for 3, 6, 14 or 30 days caused growth retardation, hepatomegaly and decreased liver lipids throughout the period of the experiment. Hypercholesterolemia was observed after feeding a histidine-excess diet for 6 days; then serum cholesterol continuously increased for 30 days. Serum triglyceride on day 30 in rats fed the histidine-excess diet showed a significant decrease compared to rats fed the basal diet. Serum phospholipids of rats fed the histidine-excess diet for 7 or 14 days showed a significant increase compared to rats fed the basal diet. When rats were fed a basal, histidine-excess or cholesterol-supplemented diet (0.5% and 1.0% cholesterol) for 6 days, the distribution of serum high density (HDL), low density (LDL) and very low density lipoprotein cholesterol in rats fed the histidine-excess diet was similar to that of rats fed the basal diet, whereas LDL-cholesterol increased and HDL-cholesterol decreased in rats fed the cholesterol-supplemented diet.     

Analysis of the stability of mutant lysozymes at position 15 using X-ray crystallography

His 15 of hen lysozyme is located at the protein surface and is partly buried by the neighboring residues. The side chain of His 15 forms hydrogen bonds with surrounding residues and these hydrogen bonds are somewhat buried. A series of mutant lysozymes at the position 15 (Gly, Ala, Val, and Phe) was prepared, and their stabilities were analyzed by GdnHCl denaturation and X-ray crystallography. The mutants were less stable than the wild type at pH 5.5 and 35 degrees C. In H15G and H15A, X-ray crystallography revealed two fixed water molecules at the mutated region, which formed similar hydrogen bonds to those in the wild type. On the other hand, it was suggested that the hydrogen bonds were disrupted and that several unfavorable van der Waals' contacts occurred in H15V and H15F. Therefore, we concluded that His 15 stabilized the lysozyme structure by forming hydrogen bonds and the best packing with the neighboring residues. Moreover, we found that the method of protein stabilization by increasing the hydrophobicity of an amino acid residue was not always effectively applicable, especially when the residue had formed a hydrogen bond.     

Simple technique for pancreatogastrostomy and the histological findings of pancreatogastric anastomosis

The leakage of pancreatic juice is the most serious complication after pancreatoduodenectomy. In an attempt to lessen the incidence of this complication, we have developed a simple technique for the anastomosis of pancreatogastrostomy. Since March 14, 1995, we have already performed surgical procedures on 8 cases of pancreatogastrostomy using this technique and as yet have never experienced fatal complications. We report the autopsy findings of pancreatogastric anastomosis histologically. The specimen taken from the anastomosis reveals good mucosal continuity. This result suggests the importance of reliable sutures between the pancreatic parenchyma and the full thickness of the gastric wall for the anastomosis of pancreatogastrostomy.