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While mating systems vary considerably throughout the animal kingdom, smell is increasingly recognised as a common factor in successful copulation. However, the full range of olfactory-in-duced mating behaviours among animals is undoubtedly underestimated in comparison to visual or contact stimuli, and this underestimation suggests that our understanding of courtship rituals may be incomplete. Here, we use the highly invasive topmouth gudgeon, Pseudorasbora parva as a biological model to separate and validate the induction of headstands as courtship behaviour in many fish species. Conspecific odour was isolated using solid phase extraction (SPE) and fractionated using high-performance liquid chromatography (HPLC). Active fractions were characterised using nuclear magnetic resonance (NMR). Electrophysiological responses of pre-ovulatory females to conspecific odour were also tested via the electro-olfactogram (EOG). Results revealed that pre-ovulatory females adopted headstand body postures during exposure to nest guarding male odour (4/12) and SPE extract (5/12) but did not display during exposure to any other odour. Fractions from HPLC yielded 1 active fraction that induced headstand body postures in pre-ovulatory females (4/12). The active fraction eluted at 30-35 min and showed no response in the UV. NMR imaging revealed low intensity levels of aliphatic protons: methylene protons (CH?) at around 2 parts per million (ppm) and methin (CH) at around 3 ppm. EOG response amplitudes were approximately double in response to pre-ovulatory female odour than to odour derived from post-ovulatory females (Mann-Whitney i7-test, p < 0.01, N = 6). This study provides evidence of sex-specific chemical cues and responsive, adaptive sexual behaviour in P. parva and demonstrates that female display postures may be induced by chemical stimuli alone. [ABSTRACT FROM AUTHOR]

Background: The simultaneous use of abdominal and endoscopic vacuum therapy in a case of an abdominal abscess caused by gastric perforation is demonstrated and innovative operative endoscopic management is described., Case Report: A computed tomography scan performed on a 67-year-old female patient showed a large abscess of the upper abdominal cavity where laparoscopic fundoplication had been performed 6 months previously. Endoscopy showed a transmural perforation of the dorsal wall of the stomach. The gastric perforation was closed and drained using intracavitary endoscopic vacuum therapy. Open pore polyurethane foam drainage was inserted through the defect into the extraluminal cavity for 3 days. A second period of therapy followed using intraluminal therapy with total drainage of the stomach, simultaneous enteral nutrition via a jejunal tube and a vacuum pressure of - 125 mmHg was applied with an electronic vacuum device. The abdominal abscess was drained via laparotomy and intra-abdominal vacuum therapy was performed with an open pore double-layered film using a vacuum pressure of - 75 mmHg. The perforation defect was not treated by operative means. Abdominal vacuum therapy ended 3 days postoperatively and the abdominal wall was closed by suture. Endoscopic vacuum therapy of the gastric perforation was terminated after 7 days and primary wound healing could then be achieved., Conclusion: Use of endoscopic and abdominal vacuum therapy as well as new open pore material is an innovative option for operative management.

In the presence of oxygen (O(2)), carbon monoxide (CO) is synthesised from heme by endogenous hemeoxygenases, and is a powerful activator of BK(Ca) channels. This transduction pathway has been proposed to contribute to cellular O(2) sensing in rat carotid body. In the present study we have explored the role that four cysteine residues (C820, C911, C995 and C1028), located in the vicinity of the "calcium bowl" of C-terminal of human BK(Ca)-alphasubunit, have on channel CO sensitivity. Mutant BK(Ca)-alphasubunits were generated by site-directed mutagenesis (single, double and triple cysteine residue substitutions with glycine residues) and were transiently transfected into HEK 293 cells before subsequent analysis in inside-out membrane patches. Potassium cyanide (KCN) completely abolished activation of wild type BK(Ca) channels by the CO donor, tricarbonyldichlororuthenium (II) dimer, at 100microM. In the absence of KCN the CO donor increased wild-type channel activity in a concentration-dependent manner, with an EC(50) of ca. 50microM. Single cysteine point mutations of residues C820, C995 and C1028 affected neither channel characteristics nor CO EC(50) values. In contrast, the CO sensitivity of the C911G mutation was significantly decreased (EC(50) ca. 100 M). Furthermore, all double and triple mutants which contained the C911G substitution exhibited reduced CO sensitivity, whilst those which did not contain this mutation displayed essentially unaltered CO EC(50) values. These data highlight that a single cysteine residue is crucial to the activation of BK(Ca) by CO. We suggest that CO may bind to this channel subunit in a manner similar to the transition metal-dependent co-ordination which is characteristic of several enzymes, such as CO dehydrogenase.

Hydrogen sulfide (H(2)S) is produced endogenously in many types of mammalian cells. Evidence is now accumulating to suggest that H(2)S is an endogenous signalling molecule, with a variety of molecular targets, including ion channels. Here, we describe the effects of H(2)S on the large conductance, calcium-sensitive potassium channel (BK(Ca)). This channel contributes to carotid body glomus cell excitability and oxygen-sensitivity. The experiments were performed on HEK 293 cells, stably expressing the human BK(Ca) channel alpha subunit, using patch-clamp in the inside-out configuration. The H(2)S donor, NaSH (100microM-10 mM), inhibited BK(Ca) channels in a concentration-dependent manner with an IC(50) of ca. 670microM. In contrast to the known effects of CO donors, the H(2)S donor maximally decreased the open state probability by over 50% and shifted the half activation voltage by more than +16mV. In addition, although 1 mM KCN completely suppressed CO-evoked channel activation, it was without effect on the H(2)S-induced channel inhibition, suggesting that the effects of CO and H(2)S were non-competitive. RT-PCR showed that mRNA for both of the H(2)S-producing enzymes, cystathionine-beta-synthase and cystathionine-gamma-lyase, were expressed in HEK 293 cells and in rat carotid body. Furthermore, immunohistochemistry was able to localise cystathionine-gamma-lyase to glomus cells, indicating that the carotid body has the endogenous capacity to produce H(2)S. In conclusion, we have shown that H(2)S and CO have opposing effects on BK(Ca)channels, suggesting that these gases have separate modes of action and that they modulate carotid body activity by binding at different motifs in the BK(Ca)alphasubunit.

The pheromone nereithione (cysteine-glutathione disulfide), which is released by swimming females of the polychaete Nereis succinea to activate spawning behavior of N. succinea males, has recently been identified and synthesized. Nereithione activates sperm release at less than 10(-6) M, one to two orders of magnitude less than oxidized glutathione or any other glutathione derivative tested. The glutathione fragment gamma-glu-cys inhibited sperm release. Nereithione aroused three components of the male nuptial behavior: circling, sperm release, and accelerated swimming. Electrophysiological activity elicited by nereithione near the sperm release site consisted of initial large spikes, cyclic bursting activity, and small spikes lasting up to a minute and was dose dependent, rapid, reversible, and repeatable. This preparation is an excellent model system for characterizing the receptors and functions of a marine pheromone.