Methane production under pressure by fermentation of waste materials

Summary Methane production by fermentation under pressure is characterised by a reduction of the gas flowrate and a simultaneous increase in the pourcentage of methane. At between O and 4 effective bars, these competing effects give a practically constant production of methane.     

Relationship of the photosensitivity of bilayer lipid membranes and the aqueous acceptor. Studies using complex ions of amino acids.

The photocurrent in photosensitive bilayer lipid membranes has been studied as a function of the aqueous acceptor. Correlations are observed between the relative photocurrent and the position of the complex ion visible absorption band and the dipole moment of the ligand. The effect of the ligands is nondirectional: they may be added to either side of the membrane with a corresponding effect on the photocurrent. The effects of the ligands are interpreted using an energy barrier model.     

Predation-dependent oviposition habitat selection by the mosquito Culiseta longiareolata: a test of competing hypotheses

We investigated the mechanism underlying oviposition habitat selection (OHS) in the mosquito Culiseta longiareolata. The putative outcome of a trade‐off between the risk of predation and detrimental density dependence, OHS in this species presents an opportunity to test two competing alternatives: (1) a polymorphic scenario, in which a fixed proportion of females constantly avoid ‘predator pools’, while the remainder oviposits at random; and (2) a monomorphic scenario, in which all females oviposit in predator pools with a certain probability. We present a conceptual framework that demonstrates how a simple experimental design – whereby predator incidence in artificial pools is alternated between 0.25 and 0.75 – can distinguish between, or refute, the two scenarios. Given the proportional use, by ovipositing females, of predator‐free pools observed under each treatment, and a bootstrap estimate of the ratio of daily oviposition rates, we find the monomorphic scenario twice as likely as the polymorphic.     

Domestication, comparative biology and interactions of wild and cultured fish: convenor's synthesis

Aquaculture is expanding rapidly and many fish species are brought into cultivation, entering a process of domestication with consequences for their morphology, physiology, ecology and evolution. In some species the abundance of cultured populations matches or exceeds that of wild stocks, and interactions between cultured and wild fish can pose significant conservation challenges. At the same time, captive breeding and re‐introduction play an important role in the conservation of some of the world's most endangered fishes. Drawing on contributions from the FSBI Symposium and the wider literature, we synthesize current knowledge of the process and extend of fish domestication, interactions between cultured and wild fish, and the use of cultured fish in fisheries enhancement and restoration. We provide a perspective on the role of biological issues within the wider context of aquaculture development and aquatic conservation biology, and conclude with a discussion of promising avenues for further research.     

Can macaroni penguins keep up with climate- and fishing-induced changes in krill?

Macaroni penguins have evolved to cope with the highly variable conditions of the Southern Ocean. However, changes in prey supply and patchiness potentially associated with changes in climate and krill fishing activity may be occurring too rapidly for the penguins to adapt. We use a stochastic dynamic programming model to examine how changes in both the mean and patchiness of krill supply may affect the foraging decisions, and therefore breeding success, of female macaroni penguins at South Georgia. We predict that rapid changes in the mean supply of prey will have more of an effect on the condition of the female and chick than changes in prey patchiness, and that changes in foraging behavior compensate for changes in prey up to a threshold point, beyond which breeding success is likely impacted. In particular, we predict that the location of the threshold is affected by whether or not the penguins are adapted to the prey environment in which they are foraging, with the female and chick receiving on average 20% less of their daily energetic requirement if the female is not foraging optimally.     

Human adenovirus proteinase: DNA binding and stimulation of proteinase activity by DNA.

The interaction of the human adenovirus proteinase (AVP) with various DNAs was characterized. AVP requires two cofactors for maximal activity, the 11-amino acid residue peptide from the C-terminus of adenovirus precursor protein pVI (pVIc) and the viral DNA. DNA binding was monitored by changes in enzyme activity or by fluorescence anisotropy. The equilibrium dissociation constants for the binding of AVP and AVP-pVIc complexes to 12-mer double-stranded (ds) DNA were 63 and 2.9 nM, respectively. DNA binding was not sequence specific; the stoichiometry of binding was proportional to the length of the DNA. Three molecules of the AVP-pVIc complex bound to 18-mer dsDNA and six molecules to 36-mer dsDNA. When AVP-pVIc complexes bound to 12-mer dsDNA, two sodium ions were displaced from the DNA. A Delta of -4.6 kcal for the nonelectrostatic free energy of binding indicated that a substantial component of the binding free energy results from nonspecific interactions between the AVP-pVIc complex and DNA. The cofactors altered the interaction of the enzyme with the fluorogenic substrate (Leu-Arg-Gly-Gly-NH)2-rhodamine. In the absence of any cofactor, the Km was 94.8 microM and the kcat was 0.002 s(-1). In the presence of adenovirus DNA, the Km decreased 10-fold and the kcat increased 11-fold. In the presence of pVIc, the Km decreased 10-fold and the kcat increased 118-fold. With both cofactors present, the kcat/Km ratio increased 34000-fold, compared to that with AVP alone. Binding to DNA was coincident with stimulation of proteinase activity by DNA. Although other proteinases have been shown to bind to DNA, stimulation of proteinase activity by DNA is unprecedented. A model is presented suggesting that AVP moves along the viral DNA looking for precursor protein cleavage sites much like RNA polymerase moves along DNA looking for a promoter.     

Interaction of the human adenovirus proteinase with its 11-amino acid cofactor pVIc

The interaction of the human adenovirus proteinase (AVP) and AVP-DNA complexes with the 11-amino acid cofactor pVIc was characterized. The equilibrium dissociation constant for the binding of pVIc to AVP was 4.4 microM. The binding of AVP to 12-mer single-stranded DNA decreased the K(d) for the binding of pVIc to AVP to 0.09 microM. The pVIc-AVP complex hydrolyzed the substrate with a Michaelis constant (K(m)) of 3.7 microM and a catalytic rate constant (k(cat)) of 1.1 s(-1). In the presence of DNA, the K(m) increased less than 2-fold, and the k(cat) increased 3-fold. Alanine-scanning mutagenesis was performed to determine the contribution of individual pVIc side chains in the binding and stimulation of AVP. Two amino acid residues, Gly1' and Phe11', were the major determinants in the binding of pVIc to AVP, while Val2' and Phe11' were the major determinants in stimulating enzyme activity. Binding of AVP to DNA greatly suppressed the effects of the alanine substitutions on the binding of mutant pVIcs to AVP. Binding of either or both of the cofactors, pVIc or the viral DNA, to AVP did not dramatically alter its secondary structure as determined by vacuum ultraviolet circular dichroism. pVIc, when added to Hep-2 cells infected with adenovirus serotype 5, inhibited the synthesis of infectious virus, presumably by prematurely activating the proteinase so that it cleaved virion precursor proteins before virion assembly, thereby aborting the infection.     

The effect of tumour bed boost on local control after breast conserving surgery. First results of the randomized boost trial of the National Institute of Oncology

PURPOSE: To evaluate the effect of tumour bed boost on local tumour control (LTC) after breast conserving surgery in a prospective study. METHODS: Between 1995 and 1998, 207 women with early invasive breast cancer who underwent conservative operation were treated by 50 Gy irradiation to the whole breast and then randomly assigned to receive either no further radiotherapy (n=103) or a boost to the tumour bed (n=104) with either 16 Gy electron (n=52) or 12-14.25 Gy high dose rate brachytherapy (n=52). RESULTS: At a median follow-up of 4.25 years the crude rate of local recurrence was 6.7% with and 13.6% without boost. The respective rates of tumour bed relapse were 3.8% vs. 10.7%. The 4 year probability of LTC, relapse-free survival and breast cancer-specific survival was 94.2% vs. 85.1% (p=0.1176), 82.3% vs. 67.2% (p=0.0438) and 84.8% vs. 90.9% (p=0.1111), respectively, in favour of the boost group. Systemic treatments had no significant impact on LTC (88.9% with and 89.6% without systemic treatment, p=0.8858). CONCLUSION: Tumour bed boost decreased the incidence of local and tumor bed relapses with a reduction of 50% and 64%, respectively. Relapse-free survival was improved significantly with boost. However, the influence of boost treatment on breast cancer-specific survival should be tested in further studies. In spite of the higher incidence of late radiation side effects in the boost arm, boost dose is strongly recommended for patients at high risk for local recurrence. The final results of the EORTC trial and other ongoing studies will help to clarify the indication of boost dose according to prognostic subgroups.     

Actin can act as a cofactor for a viral proteinase in the cleavage of the cytoskeleton

Cytoskeletal proteins are exploited by many viruses during infection. We report a novel finding that actin can act as a cofactor for the adenovirus proteinase (AVP) in the degradation of cytoskeletal proteins. Transfection studies in HeLa cells revealed AVP localized with cytokeratin 18, and this was followed by destruction of the cytokeratin network. For AVP to cleave cytokeratin 18, a cellular cofactor was shown to be required, consistent with AVP being synthesized as an inactive proteinase. Actin was considered a cellular cofactor for AVP, because the C terminus of actin is homologous to a viral cofactor for AVP. AVP was shown to bind to the C terminus of actin, and in doing so AVP exhibited full enzymatic activity. In vitro, actin was a cofactor in the cleavage of cytokeratin 18 by AVP. The proteinase alone could not cleave cytokeratin 18, but in the presence of actin, AVP cleaved cytokeratin 18. Indeed, actin itself was shown to be a cofactor and a substrate for its own destruction in that it was cleaved by AVP in vitro. Cleavage of cytoskeletal proteins weakens the structure of the cell, and therefore, actin as a cofactor may play a role in cell lysis and release of nascent virions.