Piscidin, an antibacterial peptide isolated from the mast cells of striped bass, has potent antimicrobial activity against a broad spectrum of pathogens in vitro. We investigated the mechanism of action of this 22-residue cationic peptide by carrying out structural studies and electrophysiological experiments in lipid bilayers. Circular dichroism experiments showed that piscidin was unstructured in water but had a high alpha-helix content in dodecylphosphocholine (DPC) micelles. 1H NMR data in water and TFE confirmed these results and demonstrated that the segment of residues 8-17 adopted an alpha-helical structure in a micellar environment. This molecule has a marked amphipathic character, due to well-defined hydrophobic and hydrophilic sectors. This structure is similar to those determined for other cationic peptides involved in permeabilization of the bacterial membrane. Multichannel experiments with piscidin incorporated into azolectin planar bilayers gave reproducible I-V curves at various peptide concentrations and unambiguously showed that this peptide permeabilized the membrane. This pore forming activity was confirmed by single-channel experiments, with well-defined ion channels obtained at different voltages. The characteristics of the ion channels (voltage dependence, only one or two states of conductance) clearly suggest that piscidin is more likely to permeabilize the membrane by toroidal pore formation rather than via the "barrel-stave" mechanism.

During the past 2 decades, considerable progress has been made in the study of bird semiochemistry, and our goal was to review and evaluate this literature with particular emphasis on the volatile organic constituents. Indeed, since the importance of social chemosignaling in birds is becoming more and more apparent, the search for molecules involved in chemical communication is of critical interest. These molecules can be found in different sources that include uropygial gland secretions, feather-surface compounds, and molecules from feces and skin. Although many studies have examined the chemical substances secreted by birds, research on bird chemical communication is still at the start, so new strategies for collecting samples and development of new methods of analysis are urgently required. As a first step, we built a database that brings together potential semiochemicals, using a unique chemical nomenclature for comparing different bird species and also for referencing the different classes of substances that can be found in order to adapt future parameters of analysis. The most important patterns of the wax fraction of preen secretions are highlighted and organized in an ordered table. We also draw up a list of various combinations of sampling and analytical techniques, so that each method can be compared at a glance.

SummaryMycobacteria are characterized by an unusual cell wall that controls nutrient and small hydrophilic compound permeability. Porin-like proteins are necessary to ensure the transport of molecules into the cell. Here, we investigated the pore-forming properties of OmpATb, a porin from Mycobacterium tuberculosis, in lipid bilayers. Multi-channel experiments showed an asymmetric behaviour with channel closures at negative critical voltages (Vc) and a strong decrease in Vc at acidic pH. Single-channel experiments gave conductance values of about 850 ± 80 pS in 1 M KCl and displayed a weak cationic selectivity in 4-8 pH range. The production and characterization of a series of truncated OmpATb proteins, showed that the central domain (OmpATb 73-220) was sufficient to induce the ion channel properties of the native protein in lipid bilayers, i.e. asymmetric insertion, pH-dependent voltage closure, cationic selectivity and similar conductance values in 1 M KCl. Western blot analysis suggests that the presence of OmpATb is only restricted to certain pathogenic species. Therefore, the propensity of channels of native OmpATb to close at low pH may represent an intrinsic property allowing pathogenic mycobacteria to adapt and survive to mildly acidic conditions, such as those encountered within the macrophage phagosome.

A large part of the literature on sensory perception and behavior in dolphins is devoted to its well-developed vocal and echolocation abilities. In this review, we aim to augment current knowledge by examining the literature on dolphins' entire "Merkwelt" (which refers to everything a subject perceives, creating a crucial part of the subject's Umwelt). We will show that despite extensive knowledge on audition, aspects such as context relatedness, the social function of vocalizations or socio-sexual recognition, remain poorly understood. Therefore, we propose areas for further lines of investigation. Recent studies have shown that the sensory world of dolphins might well be much more diverse than initially thought. Indeed, although underwater and aerial visual systems differ in dolphins, they have both been shown to be important. Much debated electro-and magnetoreception appear to be functional senses according to recent studies. Finally, another neglected area is chemoreception. We will summarize neuroanatomical and physiological data on olfaction and taste, as well as corresponding behavioral evidence. Taken together, we will identify a number of technical and conceptual reasons for why chemosensory data appear contradictory, which is much debated in the literature. In summary, this article aims to provide both an overview of the current knowledge on dolphin perception, but also offer a basis for further discussion and potential new lines of research.Keywords: cetaceans, Delphinidae, Tursiops truncatus, audition, vision, electroreception, magnetoreception, chemoreception DOLPHIN'S UMWELT Sensory perception is essential for the survival of organisms, be it for the detection of (un)favorable physical conditions, the presence/absence of food or predators, the detection of communication signals or the recognition of social partners. It is crucial for any species to perceive regularities and changes in the properties of their abiotic and biotic environment.

A synthetic peptide of 23 residues corresponding to the carboxyterminal 113 to 135 region of component-3 of proteose peptone (PP3) has been investigated with regard to its antibacterial properties. This cationic amphipathic peptide that we refer to as lactophoricin, displayed a growth-inhibitory activity against both gram-positive and gram-negative bacteria. For most of the strains tested, bacterial growth was observed in the presence of lactophoricin except for Streptococcus thermophilus. In that case, lactophoricin exhibited a minimum inhibitory concentration of 10 microM and a minimum lethal concentration of 20 microM. No hemolysis of human red blood cells was detected for peptide concentrations between 2 to 200 microM, indicating that lactophoricin would be noncytotoxic when used in this concentration range.

Baleen whales face the challenge of finding patchily distributed food in the open ocean. Their relatively well-developed olfactory structures suggest that they could identify the specific odours given off by planktonic prey such as krill aggregations. Like other marine predators, they may also detect dimethyl sulfide (DMS), a chemical released in areas of high marine productivity. However, dedicated behavioural studies still have to be conducted in baleen whales in order to confirm the involvement of chemoreception in their feeding ecology. We implemented 56 behavioural response experiments in humpback whales using two food-related chemical stimuli, krill extract and DMS, as well as their respective controls (orange clay and vegetable oil) in their breeding (Madagascar) and feeding grounds (Iceland and Antarctic Peninsula). The whales approached the stimulus area and stayed longer in the trial zone during krill extract trials compared to control trials, suggesting that they were attracted to the chemical source and spent time exploring its surroundings, probably in search of prey. This response was observed in Iceland, and to a lesser extend in Madagascar, but not in Antarctica. Surface behaviours indicative of sensory exploration, such as diving under the stimulus area and stopping navigation, were also observed more often during krill extract trials than during control trials. Exposure to DMS did not elicit such exploration behaviours in any of the study areas. However, acoustic analyses suggest that DMS and krill extract both modified the whales’ acoustic activity in Madagascar. Altogether, these results provide the first behavioural evidence that baleen whales actually perceive prey-derived chemical cues over distances of several hundred metres. Chemoreception, especially olfaction, could thus be used for locating prey aggregations and for navigation at sea, as it has been shown in other marine predators including seabirds.

OmpATb is the prototype of a new family of porins in Mycobacterium tuberculosis and Mycobacterium bovis BCG. Although the pore-forming activity of this protein has been clearly established by using recombinant protein produced in Escherichia coli, characterization of the native porin has been hampered by the scarce amount of protein present in the M. tuberculosis detergent extracts. To this aim, we have developed a protocol to overproduce and obtain high yields of OmpATb in both Mycobacterium smegmatis and M. bovis BCG. The protein could be extracted and purified from the cell wall fraction and subsequently used for analysis of the pore-forming activity in multichannel and single-channel conductance experiments. Our results indicate that OmpATb produced in mycobacteria presents an average conductance value of 1,600 ؎ 100 pS, slightly higher than that of OmpATb produced in E. coli, suggesting the occurrence of OmpATb in a highly ordered organization within the mycobacterial cell wall. In contrast to OmpATb, a truncated form lacking the first 72 amino acids (OmpATb 73-326 ) was essentially found in the cytosol and was not active in planar lipid bilayers. This suggested that the N-terminal domain of OmpATb could participate in targeting of OmpATb to the cell wall. This was further confirmed by analyzing M. smegmatis clones expressing a chimeric protein consisting of a fusion between the N-terminal domain of OmpATb and the E. coli PhoA reporter. The present study shows for the first time that the N terminus of OmpATb is required for targeting the porin to the cell wall and also appears to be essential for its pore-forming activity.Mycobacteria are medically important, particularly Mycobacterium tuberculosis, which causes about two millions deaths each year in the world (36). The emergence of multidrugresistant strains poses a serious threat to the treatment and control of tuberculosis. One of the main difficulties to eradicate this infection is due to the weak penetration of drugs in the cell. Mycobacteria are characterized by a highly hydrophobic cell wall in which the lipid and lipid-containing components are present in unusual abundance, constituting 60% of the dry weight of the cell wall (3). This hydrophobicity contributes to the very low permeability of the cell wall, thus preventing the diffusion of small solutes such as hydrophilic drugs (7, 13). The permeability of M. tuberculosis is comparable to that of Pseudomonas aeruginosa, whose permeability is in turn 100 times lower than that of E. coli (7).The current model of the mycobacterial cell envelope includes the presence of an outer membrane, although mycobacteria are classified as gram-positive bacteria. The outer membrane consists of mycolic acids, which are very long-chain fatty acids attached to a lower layer of arabinogalactan to form a closely packed monolayer (16). Noncovalently bound lipids complement the ordered arrangement of mycolic acids to an asymmetric bilayer (21). Despite the presence of an efficient impermeable barrier, the permeability of...

The mitochondrial outer membrane channel (VDAC), a central player in mitochondria and cell death, was reconstituted in polymer-supported phospholipid bilayers. Highly purified VDAC was first reconstituted in vesicles; channel properties and NADH-ferricyanide reductase activity were ascertained before deposition onto solid substrates. 1-Palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine/1,2-distearoyl-sn-glycero-3-phosphoethanolamine-poly(ethylene glycol)-N-hydroxysuccinimide mixed vesicles containing VDAC were linked onto amine-grafted surfaces (glass and gold) and disrupted to form a VDAC-containing polymer-tethered planar bilayer. Surface plasmon spectroscopy, fluorescence microscopy, and atomic force microscopy measurements ascertained the membrane thickness, fluidity, and continuity. VDAC reconstituted in bilayers efficiently transported calcium ions and was modulable by two channel blockers, 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid and l-glutamate. The novel setup may allow the study of the assembly of a polyprotein complex centered on VDAC and its role in mitochondrial biology, calcium fluxes, and apoptosis.