Role of Inorganic and Organic Fractions in Animal Manure Compost in Lead Immobilization and Microbial Activity in Soil

This study aimed to identify how the ratio of inorganic-to-organic components in animal manure compost (AMC) affected both lead immobilization and microbial activity in lead-contaminated soil. When AMC containing 50% or more inorganic fraction with high phosphorous content was applied to contaminated soil, the amounts of water-soluble lead in it were suppressed by over 88% from the values in the soil without compost. The residual fraction under sequential extraction increased with the inorganic fraction in the AMC; however, in those AMCs, the levels of microbial enzyme activity were the same or less than those in the control soil. The application of AMC containing 25% inorganic fraction could alter the lead phases to be more insoluble while improving microbial enzyme activities; however, no suppression of the level of water-soluble lead existed during the first 30 days. These results indicate that compost containing an inorganic component of 50% or more with high phosphorus content is suitable for immobilizing lead; however, in the case where low precipitation is expected for a month, AMC containing 25% inorganic component could be used to both immobilize lead and restore microbial activity

Candida albicans Possesses Sap7 as a Pepstatin A-Insensitive Secreted Aspartic Protease

BACKGROUND: Candida albicans, a commensal organism, is a part of the normal flora of healthy individuals. However, once the host immunity is compromised, C. albicans opportunistically causes recurrent superficial or fatal systemic candidiasis. Secreted aspartic proteases (Sap), encoded by 10 types of SAP genes, have been suggested to contribute to various virulence processes. Thus, it is important to elucidate their biochemical properties for better understanding of the molecular mechanisms that how Sap isozymes damage host tissues. METHODOLOGY/PRINCIPAL FINDINGS: The SAP7 gene was cloned from C. albicans SC5314 and heterogeneously produced by Pichia pastoris. Measurement of Sap7 proteolytic activity using the FRETS-25Ala library showed that Sap7 was a pepstatin A-insensitive protease. To understand why Sap7 was insensitive to pepstatin A, alanine substitution mutants of Sap7 were constructed. We found that M242A and T467A mutants had normal proteolytic activity and sensitivity to pepstatin A. M242 and T467 were located in close proximity to the entrance to an active site, and alanine substitution at these positions widened the entrance. Our results suggest that this alteration might allow increased accessibility of pepstatin A to the active site. This inference was supported by the observation that the T467A mutant has stronger proteolytic activity than the wild type. CONCLUSIONS/SIGNIFICANCE: We found that Sap7 was a pepstatin A-insensitive protease, and that M242 and T467 restricted the accessibility of pepstatin A to the active site. This finding will lead to the development of a novel protease inhibitor beyond pepstatin A. Such a novel inhibitor will be an important research tool as well as pharmaceutical agent for patients suffering from candidiasis

Preparation of Mesoporous Bimetallic Au-Pt with a Phase-Segregated Heterostructure Using Mesoporous Silica

Mesoporous bimetallic Au-Pt with a phase-segregated heterostructure has been prepared by using mesoporous silica SBA-15 as a template. Au nanoparticles were prepared as a seed metal within the mesopores, and subsequently Pt was deposited, sandwiching the Au seeds. Energy-dispersive X-ray (EDX) spectral mapping showed that the framework of mesoporous bimetallic Au-Pt, prepared by removing the silica template with HF, was composed of Au nanoparticles joined with Pt nanowires. The Au/Pt ratio of the mesoporous bimetallic Au-Pt could be varied by controlling the number of Au deposition cycles. Pre-adsorbed CO (COad) stripping voltammetry of the mesoporous bimetallic Au-Pt showed that the surfaces of the joined bimetallic structure were electrochemically active. This could be attributed to the open framework structure having a high ratio of exposed bimetallic mesopore surfaces. The described preparative approach, involving a mesoporous silica template and stepwise deposition within the mesopores, enables control of the nanostructure of the bimetallic material, which is greatly promising for the further development of synthetic methodologies for bimetallic structures

Additional file 5: of Description of the interaction between Candida albicans and macrophages by mixed and quantitative proteome analysis without isolation

Proteins identified in this study for C. albicans monoculture (976 proteins) and macrophage monocultre (1,357 proteins)

Protein sequence alignment between Sap1–3 and Sap7.

<p>Multiple-sequence alignments were performed with CLUSTAL W. The active site of Sap7 was predicted to be D244 and D464 by homology. The cleavage point of Sap7 was found in the Sap7-specific insertion sequence N422-N451, which did not exist in the sequences of Sap1–3.</p

Additional file 7: of Description of the interaction between Candida albicans and macrophages by mixed and quantitative proteome analysis without isolation

Changed proteins of macrophage interacting with C. albicans (five proteins)