Cloning and disruption of the antigenic catalase gene of Aspergillus fumigatus

Aspergillus fumigatus possesses two catalases (described as fast and slow on the basis of their electrophoretic mobility). The slow catalase has been recognized as a diagnostic antigen for aspergillosis in immunocompetent patients. The antigenic catalase has been purified. The enzyme is a tetrameric protein composed of 90-kDa subunits. The corresponding cat1 gene was cloned, and sequencing data show that the cat1 gene codes for a 728-amino-acid polypeptide. A recombinant protein expressed in Pichia pastoris is enzymatically active and has biochemical and antigenic properties that are similar to those of the wild-type catalase. Molecular experiments reveal that CAT1 contains a signal peptide and a propeptide of 15 and 12 amino acid residues, respectively. cat1-disrupted mutants that were unable to produce the slow catalase were as sensitive to H2O2 and polymorphonuclear cells as the wild-type strain. In addition, there was no difference in pathogenicity between the cat1 mutant and its parental cat1+ strain in a murine model of aspergillosis.     

Elimination of organic matter present in wastewaters from the cork industry by membrane filtration

BACKGROUND: The first stage of the cork industrial process generates great volumes of wastewater with moderate to high organic pollutant content that must be purified using different procedures, such as filtration by membranes. RESULTS: The tangential filtration of these wastewaters was studied using two different laboratory equipments. In the first one, three ultrafiltration (UF) membranes were tested, with molecular weight cut‐off (MWCO) 100 kDa and 30 kDa, and two operating modes were used: total recycling of permeate and retentate streams, and in continuous mode, without recycling both streams. In the total recycling UF experiments, the influence of the operating variables on the permeate flux was first established. The effectiveness of the different membranes was determined by evaluating the rejection coefficients for several parameters that measure the global pollutant content of the effluent. The values found for these rejection coefficients were in the following order: ellagic acid and color > absorbance at 254 nm > tannic content > COD (chemical oxygen demand). In the continuous mode experiments, the fouling mechanism for each membrane was established by fitting the experimental data to various filtration fouling models given in the literature. The operating mode in the second equipment was batch concentration, and additional experiments were carried out with an UF membrane (2 kDa), and with a NF membrane (with MWCO in the range 150–300 Da). CONCLUSIONS: The three operating modes tested provided different rejection levels of organic matter; among them, the most effective procedure tested was batch concentration mode using a NF membrane. Copyright © 2007 Society of Chemical Industry     

Immunoelectron localization of mitochondrial F1 factor in cryosections of heart muscle cells

Immunocytochemistry was used to investigate the localization of F1 ATPase in mitochondria of cryosections of adult mouse heart muscle cells. The initial aldehyde fixation was the only denaturation step for antigens. The fine structure was preserved with contrast enhancement as the sections were maintained hydrated, with the advantage that the entire procedure is completed in one working day. The reaction was highly specific, and entire mitochondria were labeled with the Protein A-gold complex. A new analytical technique, electron spectroscopic imaging (ESI), contributed to a better visualization of the localization of the F1 factor.     

Purification of Ellagic Acid by UF Membranes

Ellagic acid (2, 3, 7, 8‐tetrahydroxy(1)benzopyrano(5, 4, 3‐cde)(1)benzopyran‐5, 10‐dione) was selected as a model pollutant which is present in the tannic fraction of cork processing wastewater. The ultrafiltration of aqueous ellagic acid solutions through three membranes was studied in tangential UF laboratory equipment. Two of the membranes were polyethersulfone (Biomax10K and Biomax5K, with MWCO of 10000 and 5000 Da, respectively), and the third made of regenerated cellulose (Ultracel5K, with MWCO of 5000 Da). The water hydraulic permeability was evaluated for each membrane. The evolution of the permeate flow rate with processing time was followed, and the influence of the main operating variables (feed flow rate, trans‐membrane pressure and nature of the membranes) on the permeate flux was also established. According to the hypothesis of the film theory, the intrinsic and apparent rejection coefficients, as well as the mass transfer coefficients, were also determined, and the values obtained were discussed as a function of the operating conditions used.     

2,4,6‐Trichlorophenol and phenol removal in methanogenic and partially‐aerated methanogenic conditions in a fluidized bed bioreactor

A fluidized bed bioreactor (FBBR) was operated for more than 575 days to remove 2,4,6‐trichlorophenol (TCP) and phenol (Phe) from a synthetic toxic wastewater containing 80 mg L^?1 of TCP and 20 mg L^?1 of Phe under two regimes: Methanogenic (M) and Partially‐Aerated Methanogenic (PAM). The mesophilic, laboratory‐scale FBBR consisted of a glass column (3 L capacity) loaded with 1 L of 1 mm diameter granular activated carbon colonized by an anaerobic consortium. Sucrose (1 g COD L^?1) was used as co‐substrate in the two conditions. The hydraulic residence time was kept constant at 1 day. Both conditions showed similar TCP and Phe removal (99.9 + %); nevertheless, in the Methanogenic regime, the accumulation of 4‐chlorophenol (4CP) up to 16 mg L^?1 and phenol up to 4 mg L^?1 was observed, whereas in PAM conditions 4CP and other intermediates were not detected. The specific methanogenic activity of biomass decreased from 1.01 ± 0.14 in M conditions to 0.19 ± 0.06 mmolCH₄ h^?1 gTKN^?1 in PAM conditions whereas the specific oxygen uptake rate increased from 0.039 ± 0.008 in M conditions to 0.054 ± 0.012 mmolO₂ h^?1 gTKN^?1, which suggested the co‐existence of both methanogenic archaea and aerobic bacteria in the undefined consortium. The advantage of the PAM condition over the M regime is that it provides for the thorough removal of less‐substituted chlorophenols produced by the reductive dehalogenation of TCP rather than the removal of the parent compound itself. Copyright © 2005 Society of Chemical Industry     

Characterisation and biological treatment of greywater.

Characterisation of greywater was conducted in two different greywater streams in the Netherlands (Groningen and Sneek). The concentrations of macropollutants and nutrients measured were very different in both streams; in particular the COD was 425 mg/L in Groningen's water whereas in Sneek it was 1,583 mg/L. The aerobic treatment of greywater in a fed-batch reactor led to a 90% removal of COD at different organic loading rates. Anaerobically, the removal reached 40% COD removal on average, the possible reason being the high amount of surfactants present in the influent.     

Ethics is fragile,goodness is not

This paper first illustrates what kind of ethical issues arise from the new information, communication and automation technology. It then argues that we may embrace the popular idea that technology is ethically neutral or even ambivalent without having to close our eyes to those issues and in fact, that the ethical neutrality of technology makes them all the more urgent. Finally, it suggests that the widely ignored fact of normal responsible behaviour offers a new and fruitful starting point for any future thinking about such issues.     

Effect of various parameters on sodium dodecyl sulfate (SDS) flux through a collagen membrane

The influence of various experimental parameters on the flux of sodium dodecyl sulfate (SDS) through a collagen membrane has been studied. The variables evaluated were donor concentration, time, temperature, pH and ionic strength. Data on the influence of both surfactant concentration and time on surfactant diffusion through the collagen film allow one to postulate a diffusion process mainly of the monomeric type. This diffusion mechanism based on surfactant monomers has been corroborated by studying the effect of ionic strength. Thisin vitro technique could be a useful tool to predict the effect of diverse experimental parameters on the percutaneous absorption of surfactants.