Regulatory and structural properties differentiating the chromosomal and the bacteriophage-associated <Emphasis Type="Italic">Escherichia coli</Emphasis> O157:H7 Cu,Zn Superoxide Dismutases

Background  

Highly virulent enterohemorrhagic Escherichia coli O157:H7 strains possess three sodC genes encoding for periplasmic Cu, Zn superoxide dismutases: sodC, which is identical to the gene present in non-pathogenic E. coli strains, and sodC-F1 and sodC-F2, two nearly identical genes located within lambdoid prophage sequences. The significance of this apparent sodC redundancy in E. coli O157:H7 has not yet been investigated.     

“Density equilibrium” method for the quantitative and rapid in situ determination of lipid,hydrocarbon, or biopolymer content in microorganisms

The work provides a simple method, based on a direct density equilibrium measurement, for the rapid in situ estimation of total lipid, hydrocarbon or biopolymer content in a variety of prokaryotic and eukaryotic samples. The method can be readily applied to live microalgae and photosynthetic bacteria, single‐celled or colonial microorganisms, as well as cellular fractions and isolated subcellular compartments or components. In this approach, the absolute lipid, hydrocarbon, or biopolymer content of the cells can be readily calculated. This method is especially useful for tracking the oil or polymer content of strains of microalgae and other microorganisms, whose lipid, hydrocarbon or biopolymer content may change with cultivation conditions and/or time, as the case would be in microorganism lipid‐induction industrial processes. The method is also useful for the direct in situ measurement of storage polymer accumulation in live cells, such as starch in microalgae and polyhydroxybutyrate, or other polyhydroxyalkanoates, in photosynthetic and non‐photosynthetic bacteria. Biotechnol. Bioeng. 2009;102: 1406–1415. © 2008 Wiley Periodicals, Inc.     

Solar energy conversion efficiencies in photosynthesis: Minimizing the chlorophyll antennae to maximize efficiency

The theoretical maxima of solar energy conversion efficiencies and productivities in oxygenic photosynthesis are evaluated. These are contrasted with actual measurements in a variety of photosynthetic organisms, including green microalgae, cyanobacteria, C4 and C3 plants. Minimizing, or truncating, the chlorophyll antenna size of the photosystems can improve photosynthetic solar energy conversion efficiency and productivity up to 3-fold. Generation of truncated light-harvesting chlorophyll antenna size (tla) strains, in all classes of photosynthetic organisms would help to alleviate excess absorption of sunlight and the ensuing wasteful dissipation of excitation energy, and to maximize solar-to-product energy conversion efficiency and photosynthetic productivity in high-density mass cultivations. The tla concept may find application in the commercial exploitation of microalgae and plants for the generation of biomass, biofuels, chemical feedstocks, as well as nutraceuticals and pharmaceuticals.     

Chimpanzees coordinate in a negotiation game

A crucially important aspect of human cooperation is the ability to negotiate to cooperative outcomes when interests over resources conflict. Although chimpanzees and other social species may negotiate conflicting interests regarding travel direction or activity timing, very little is known about their ability to negotiate conflicting preferences over food. In the current study, we presented pairs of chimpanzees with a choice between two cooperative tasks—one with equal payoffs (e.g., 5-5) and one with unequal payoffs (higher and lower than in the equal option, e.g., 10-1). This created a conflict of interests between partners with failure to work together on the same cooperative task resulting in no payoff for either partner. The chimpanzee pairs cooperated successfully in as many as 78–94% of the trials across experiments. Even though dominant chimpanzees preferred the unequal option (as they would obtain the largest payoff), subordinate chimpanzees were able to get their way (the equal option) in 22–56% of trials across conditions. Various analyses showed that subjects were both strategic and also cognizant of the strategies used by their partners. These results demonstrate that one of our two closest primate relatives, the chimpanzee, can settle conflicts of interest over resources in mutually satisfying ways—even without the social norms of equity, planned strategies of reciprocity, and the complex communication characteristic of human negotiation.     

Biophysical mechanisms of endotoxin neutralization by cationic amphiphilic peptides

Bacterial endotoxins (lipopolysaccharides (LPS)) are strong elicitors of the human immune system by interacting with serum and membrane proteins such as lipopolysaccharide-binding protein (LBP) and CD14 with high specificity. At LPS concentrations as low as 0.3 ng/ml, such interactions may lead to severe pathophysiological effects, including sepsis and septic shock. One approach to inhibit an uncontrolled inflammatory reaction is the use of appropriate polycationic and amphiphilic antimicrobial peptides, here called synthetic anti-LPS peptides (SALPs). We designed various SALP structures and investigated their ability to inhibit LPS-induced cytokine secretion in vitro, their protective effect in a mouse model of sepsis, and their cytotoxicity in physiological human cells. Using a variety of biophysical techniques, we investigated selected SALPs with considerable differences in their biological responses to characterize and understand the mechanism of LPS inactivation by SALPs. Our investigations show that neutralization of LPS by peptides is associated with a fluidization of the LPS acyl chains, a strong exothermic Coulomb interaction between the two compounds, and a drastic change of the LPS aggregate type from cubic into multilamellar, with an increase in the aggregate sizes, inhibiting the binding of LBP and other mammalian proteins to the endotoxin. At the same time, peptide binding to phospholipids of human origin (e.g., phosphatidylcholine) does not cause essential structural changes, such as changes in membrane fluidity and bilayer structure. The absence of cytotoxicity is explained by the high specificity of the interaction of the peptides with LPS.     

Hydroxytyrosol glucuronides protect renal tubular epithelial cells against H(2)O(2) induced oxidative damage

Hydroxytyrosol (2-(3′,4′-dihydroxyphenyl)ethanol; HT), the most active ortho-diphenolic compound, present either in free or esterified form in extravirgin olive oil, is extensively metabolized in vivo mainly to O-methylated, O-sulfated and glucuronide metabolites. We investigated the capacity of three glucuronide metabolites of HT, 3′-O-β-d-glucuronide and 4′-O-β-d-glucuronide derivatives and 2-(3′,4′-dihydroxyphenyl)ethanol-1-O-β-d-glucuronide, in comparison with the parent compound, to inhibit H₂O₂ induced oxidative damage and cell death in LLC-PK1 cells, a porcine kidney epithelial cell line. H₂O₂ treatment exerted a toxic effect inducing cell death, interacting selectively within the pro-death extracellular-signal relate kinase (ERK 1/2) and the pro-survival Akt/PKB signaling pathways. It also produced direct oxidative damage initiating the membrane lipid peroxidation process. None of the tested glucuronides exhibited any protection against the loss in renal cell viability. They also failed to prevent the changes in the phosphorylation states of ERK and Akt, probably reflecting their inability to enter the cells, while HT was highly effective. Notably, pretreatment with glucuronides exerted a protective effect at the highest concentration tested against membrane oxidative damage, comparable to that of HT: the formation of malondialdehyde, fatty acid hydroperoxides and 7-ketocholesterol was significantly inhibited.     

Coordination of Chimpanzees (<Emphasis Type="Italic">Pan troglodytes</Emphasis>) in a Stag Hunt Game

Group-living animals frequently face situations in which they must coordinate individual and sometimes conflicting goals. We assessed chimpanzees’ ability to coordinate in a Stag Hunt game. Dyads were confronted with a situation in which each individual was already foraging on a low-value food (hare) when a high-value food (stag) appeared that required collaboration for retrieval, with a solo attempt to get the stag resulting in a loss of both options. In one condition visibility between partners was open whereas in the other it was blocked by a barrier. Regardless of condition, dyads almost always (91%) coordinated to choose the higher valued collaborative option. Intentional communication or monitoring of the partner’s behavior before decision making—characteristic of much human coordination—were limited. Instead, all dyads adopted a leader–follower strategy in which one partner took the risk of going first, presumably predicting that this would induce the other to join in (sometimes communicating if she was slow to do so). These results show that humans’ closest primate relatives do not use complex communication to coordinate but most often use a less cognitively complex strategy that achieves the same end.     

Development of Mice with Brain-Specific Deletion of Floxed Glud1 (Glutamate Dehydrogenase 1) Using Cre Recombinase Driven by the Nestin Promoter

In the brain, Glud1-encoded glutamate dehydrogenase plays a major role in the recycling of the neurotransmitter glutamate. We recently reported a new model of brain-specific Glud1 null mice (Cns-Glud1 ^?/?) lacking glutamate dehydrogenase in the central nervous system. Cns-Glud1 ^?/? mice exhibit reduced astrocytic glutamate breakdown and redirection of glutamate pathways without altering synaptic transmission. Cns-Glud1 ^?/? mice were generated using LoxP and Nestin-Cre technology. Nestin-Cre mice are widely used to investigate gene deletion in the central nervous system. However, the Nes-Cre transgene itself was reported to induce a phenotype related to body weight gain. Here, we review the potential side-effects of Nes-Cre and analysed Cns-Glud1 ^?/? body weight gain. Overall, Nestin-Cre mice may exhibit transient and moderate growth retardation during the few weeks immediately following weaning. Pending appropriate controls and homogenization of the genetic background, Nestin-Cre technology is a valuable tool enabling disruption of genes of interest in the central nervous system.