Endothelial cell-specific ETB receptor knockout: autoradiographic and histological characterisation and crucial role in the clearance of endothelin-1.

Inactivation of endothelin B receptors (ETB), either through selective pharmacological antagonism or genetic mutation, increases the circulating concentration of endothelin-1 (ET-1), suggesting ETB plays an important role in clearance of this peptide. However, the cellular site of ETB-mediated clearance has not yet been determined. We have used a novel mouse model of endothelial cell-specific knockout (KO) of ETB (EC ETB(-/-)) to evaluate the relative contribution of EC-ETB to the clearance of ET-1. Phenotypic evidence of EC-specific ETB KO was confirmed by immunocytochemistry and autoradiography. Binding of the radiolabelled selective ETB ligand BQ3020 was significantly and selectively decreased in EC-rich tissues of EC ETB(-/-) mice, including the lung, liver, and kidney. By contrast, ETA binding was unaltered. RT-PCR confirmed equal expression of ET-1 in tissue from EC ETB(-/-) mice and controls, despite increased concentration of plasma ET-1 in EC ETB(-/-). Clearance of an intravenous bolus of [(125)I]ET-1 was impaired in EC ETB(-/-) mice. Pretreatment with the selective ETB antagonist A192621 impaired [(125)I]ET-1 clearance in control animals to a similar extent, but did not further impair clearance in EC ETB(-/-) mice. These studies suggest that EC-ETB are largely responsible for the clearance of ET-1 from the circulation.

Fish-habitat associations on a subsea pipeline within an Australian Marine Park.

Subsea pipelines have been installed in all major hydrocarbon basins across the globe to support the offshore Oil & Gas (O&G) industry. These artificial structures provide hard substratum that can be colonised and utilised by sessile and mobile organisms. The present study utilises industry-collected remotely operated vehicle (ROV) video to assess fish species richness and abundance, and marine growth type, extent and complexity along sections of a subsea gas pipeline, in 56-82 m depth, that traverses the Australian Commonwealth Montebello Marine Park (MMP). A total of 7493 fish from 81 species and 33 families were recorded from 606 analysed 10 m transects spaced across sections of the pipeline. Of these 81 species, 27 are considered fishery-target species in the Pilbara Demersal Scalefish fishery (PDSF), with select commercial fishing activities permitted with authorisation within the Marine Park. A moderate abundance (175) of sub-adult red emperor (Lutjanus sebae), a fishery-indicator species, were observed along the pipeline. Eleven different categories of marine growth habitat were observed, with the pipeline possessing quite uniform coverage of encrusting marine growth (coralline algae, bryozoans, ascidians, etc.) with patchy occurrences of more structurally complex sponges and black/octocoral forms. Fish species richness and abundance of the commercially targeted Moses' snapper (Lutjanus russellii) were correlated positively with increasing cover of sponges. The pipeline itself had very few spans and was never more than fractionally buried. Despite the somewhat homogenous habitats, depths, and position of the pipeline relative to the seafloor, presence of a field joint indent had a positive influence on the abundance of some common and commercially important fish species. This study demonstrates the ecological value of ROV footage obtained during industry inspection operations that were conducted for reasons unrelated to the determination of ecological information. The pipeline offers a corridor of hard bottom habitat within a marine park that facilitates epibiotic growth and the presence of reef-associated species in a region characterised by sandy sediments. Results indicate the potential importance of subsea O&G infrastructure as a habitat for fish, and in consequence, potentially also as structures with value to fisheries.

Pierce's Disease Bacterium: Mechanism of Transmission by Leafhopper Vectors.

The bacterium that causes Pierce's disease of grapevines is isolated most consistently from the foregut of its leafhopper vector Graphocephala atropunctata. As seen in light and scanning electron microscopy of infective leafhoppers, the bacteria are attached to the cibarial pump and the lining of the esophagus in the foregut where they appear to multiply. These findings suggest that the bacterium is transmitted from the foregut by egestion during feeding by infective leafloppers.

Fish associated with a subsea pipeline and adjacent seafloor of the North West Shelf of Western Australia.

Information on the potential ecological value of offshore oil and gas infrastructure is required as it reaches the end of its operational life and decisions must be made regarding the best practice option for decommissioning. This study uses baited remote underwater stereo-video systems to assess fish assemblages along an offshore subsea pipeline and in adjacent natural seabed habitats at ∼140 m depth on the North West Shelf of Western Australia. A total of 955 fish from 40 species and 25 families were recorded. Species richness was, on average 25% higher on the pipeline (6.48 ±â€¯0.37 SE) than off (4.81 ±â€¯0.28 SE) while relative abundance of fish was nearly double on the pipeline (20.38 ±â€¯2.81 SE) than in adjacent natural habitats (10.97 ±â€¯1.02 SE). The pipeline was characterised by large, commercially important species known to associate with complex epibenthic habitat and, as such, possessed a biomass of commercial fish ca 7.5 × higher and catch value ca. 8.6 × ($65.11 ±â€¯$11.14 SE) than in adjacent natural habitats ($7.57 ±â€¯$2.41 SE). This study has added to the knowledge of fish assemblage associations with subsea infrastructure and provides a greater understanding of the ecological and fisheries implications of decommissioning, helping to better inform decision-making on the fate of infrastructure.

Spatiotemporal pattern of nicotinamide adenine dinucleotide phosphate-diaphorase reactivity in the developing central nervous system of premetamorphic Xenopus laevis tadpoles.

We have catalogued the progressive appearance of putative nitric oxide synthase (NOS)-containing neurons in the developing central nervous system (CNS) of Xenopus laevis. Xenopus embryos and larvae were processed in wholemount and in cross section using nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) histochemistry as a marker for NOS within the CNS. The temporal sequence of NADPH-d reactivity identified discrete groups and subgroups of neurons in the forebrain, midbrain, and hindbrain on the basis of their morphology, location, and order of appearance during development. A proportion of these groups of neurons appeared to be important in sensory processing and motor control. Staining also appeared at specific stages in the spinal cord, the retina, and the skin. After the appearance of labelling, NADPH-d reactivity continued in each of the cell groups throughout the stages examined. We found no evidence for staining that subsequently disappeared at later stages in any cell group, indicating a persistent rather than transient role for NO in the Xenopus tadpole CNS. These results are discussed in light of recent findings on possible roles for NADPH-d-positive cell groups within the developing motor circuitry.

Serum lipid profile determines platelet reactivity to native and modified LDL-cholesterol in humans.

The effects of thrombin (0.2 U/ml) and native (n-LDL), malondialdehyde-modified (MDA-LDL) and auto-oxidized (ox-LDL) low-density lipoproteins (20 micrograms of protein/ml) on platelet activation were evaluated in seven hyperlipidemic patients and compared to seven controls (fasting serum cholesterol 8.49 +/- 0.5 and 4.61 +/- 0.4 mM, respectively). Basal and thrombin-induced increases in platelet intracellular free calcium ion concentration ([Ca2+]i; fura-2) were similar in hyperlipidemic patients and controls (45 +/- 5 vs 42 +/- 3 and 635 +/- 51 vs 599 +/- 69 mM, respectively). n-LDL, MDA-LDL and ox-LDL increased basal [Ca2+]i (16, 36 and 81 percent, p < 0.01 between LDL-types), increases were consistently smaller in patients. There was an inverse relationship between LDL-induced responses and fasting serum LDL cholesterol as well as LDL/HDL ratio. In conclusion, modified LDL activated platelets to a greater extent than n-LDL, suggesting different types of LDL-receptors. Their agonistic effect was inversely related to the fasting serum lipid profile, suggesting that blunting of platelet responses to LDL could represent a protective mechanism in hyperlipidemic patients.

The development of neuromodulatory systems and the maturation of motor patterns in amphibian tadpoles.

The relative simplicity of the amphibian tadpole nervous system has been utilised as a model for the mechanisms underlying the generation and development of vertebrate locomotion. In this paper, we review evidence on the role of descending brainstem projections in the maturation and intrinsic modulation of tadpole spinal motor networks. Three transmitter systems that have been investigated utilise the biogenic amines serotonin (5HT) and noradrenaline (NA) and the inhibitory amino acid gamma-aminobutyric acid (GABA). The distribution, development and spinal targets of these systems will be reviewed. More recent data on the role of nitric oxide (NO) will also be discussed. This ubiquitous gaseous signalling molecule is known to play a crucial role in the developing nervous system, but until recently, had not been directly implicated in the brain regions involved in motor control. NO appears to be produced by three homologous brainstem clusters in the developing motor networks of two closely related amphibian species, Xenopus laevis and Rana temporaria but, surprisingly, it plays contrasting roles in these species. Given the presumed co-localisation and interaction of nitric oxide with conventional neurotransmitters, we discuss the potential relationship of nitrergic neurons with 5HT, NA and GABA in these amphibian models.

Self-administration of medical modalities (SAMM): another method of rehabilitation education.

In order to train patients to carry out home pulmonary care adequately, we developed a hospital-based patient-education program we call Self-Administration of Medical Modalities (SAMM). This teaches patients about their pulmonary disease; about their medications' purposes, side effects and what to do if they occur, possible conflict with other medications, and the medication schedule; about use, care, and cleaning of aerosol inhalation devices and scheduling of aerosol medication treatments; and about chest physical therapy if it is indicated. Nurses, respiratory therapists, and physical therapists in the hospital teach and reinforce these concepts and evaluate the patient's progress in learning. The patient advances through three levels of competency. At Level I he is responsible only for keeping track of his medication and treatment schedules. At Level II the patient initiates requests for medication and treatments on schedule, takes them under supervision, and makes a written record of having done so. At Level III the patient's medications are kept at his bedside, he prepares and takes the medications himself, takes treatments himself, and he keeps written records. At this level the program simulates home conditions as much as possible. Patients have reported that they liked administering their own medications and treatments and that the SAMM Program was helpful in preparing them for self-care at home.

Divergent actions of serotonin receptor activation during fictive swimming in frog embryos.

We have investigated the pharmacology underlying locomotor system responses to serotonin (5-HT) in embryos of the frog, Rana temporaria, to provide a comparison to studies in embryos of its close relative, Xenopus laevis. Our findings suggest that two divergent mechanisms underlie the modulation of locomotion by 5-HT in Rana. Bath-applied 5-HT or 5-carboxamidotyptamine, a 5-HT(1,5A,7) receptor agonist, can modulate fictive swimming in a dose-dependent manner, increasing burst durations and cycle periods. However, activation of 5-HT(1,7) receptors with R8-OHDPAT or 8-OHDPAT fails to mimic 5-HT, and in some cases exerts exactly the opposite response; decreasing burst durations and cycle periods. Elevating endogenous 5-HT levels by blocking re-uptake with clomipramine transiently increases burst durations. The receptors involved in this endogenous response include 5-HT(1A) receptors, as in Xenopus, but also 5-HT(7) receptors. However, like the 8-OHDPAT enantiomers, prolonged re-uptake inhibition can result in a motor response in the opposite direction to exogenous 5-HT. This effect is not reversed by 5-HT(1A) and/or 5-HT(7) receptor antagonism, implicating 5-HT(1B/1D) receptors. Remarkably, antagonism of these receptors using methiothepin unmasks a dose-dependent response to clomipramine, reminiscent of exogenous 5-HT. Our data suggest that 5-HT(1A,7) and 5-HT(1B/1D) receptors act as gain-setters of burst durations, whilst 5-HT(5A) receptors are involved in the effects of bath-applied 5-HT on locomotion.

Observation of symbiote migration in human body lice with scanning and transmission electron microscopy.

Bacterial symbiotes in the human body louse Pediculus humanus migrate from the mycetome to the lateral oviducts during the adult molt. Their migration was first described by Ries (E. Ries. 1931. Z. Morphol. Oekol. Tiere, 20:233-367.), who examined sectioned specimens with light microscopy. The present study is a more detailed investigation which involves the use of scanning and transmission electron micrographs. The results of our studies confirm Ries' observations. Micrographs are presented of symbiotes emerging from the mycetome, migrating to the reproductive tract, and invading the lateral oviducts.

The distribution of NADPH-diaphorase-labelled interneurons and the role of nitric oxide in the swimming system of Xenopus laevis larvae.

The possible involvement of the free radical gas nitric oxide (NO) in the modulation of spinal rhythm-generating networks has been studied using Xenopus laevis larvae. Using NADPH-diaphorase histochemistry, three putative populations of nitric oxide synthase (NOS)-containing cells were identified in the brainstem. The position and morphology of the largest and most caudal population suggested that a proportion of these neurons is reticulospinal. The possible contribution of nitrergic neurons to the control of swimming activity was examined by manipulating exogenous and endogenous NO concentrations in vivo with an NO donor (SNAP, 100-500 micromol l(-)(1)) and NOS inhibitors (l-NAME and l-NNA, 0.5-5 mmol l(-)(1)), respectively. In the presence of SNAP, swim episode duration decreased and cycle period increased, whereas the NOS inhibitors had the opposite effects. We conclude from these data that the endogenous release of NO from brainstem neurons extrinsic to the spinal cord of Xenopus laevis larvae exerts a continuous modulatory influence on swimming activity, functioning like a 'brake'. Although the exact level at which NO impinges upon the swimming rhythm generator has yet to be determined, the predominantly inhibitory effect of NO suggests that the underlying mechanisms of NO action could involve modulation of synaptic transmission and/or direct effects on neuronal membrane properties.

Pea aphid symbiont relationships established by analysis of 16S rRNAs.

The pea aphid (Acyrthosiphon pisum Harris) harbors two morphologically distinct procaryotic intracellular symbionts. The genes for the 16S rRNA from these symbionts have been cloned and sequenced. Comparisons with sequences of 16S rRNAs from selected procaryotes indicate that the two symbionts are evolutionarily distinct from each other and are members of the gamma-3 subdivision of the class Proteobacteria. One of the symbionts is a member of the family Enterobacteriaceae, while the other constitutes a lineage distinct from these organisms. Both symbionts appear to have only one copy of their rRNA operon.

Induction of a non-rhythmic motor pattern by nitric oxide in hatchling Rana temporaria embryos.

Nitric oxide (NO) is a ubiquitous neuromodulator with a diverse array of functions in a variety of brain regions, but a role for NO in the generation of locomotor activity has yet to be demonstrated. The possibility that NO is involved in the generation of motor activity in embryos of the frog Rana temporaria was investigated using the NO donors S-nitroso-n-acetylpenicillamine (SNAP; 100--500 micromol l(-1)) and diethylamine nitric oxide complex sodium (DEANO; 25--100 micromol l(-1)). Immobilised Rana temporaria embryos generate a non-rhythmic 'lashing' motor pattern either spontaneously or in response to dimming of the experimental bath illumination. Bath-applied NO donors triggered a qualitatively similar motor pattern in which non-rhythmic motor bursts were generated contra- and ipsilaterally down the length of the body. The inactive precursor of SNAP, n-acetyl-penicillamine (NAP), at equivalent concentrations did not trigger motor activity. NO donors failed to initiate swimming and had no measurable effects on the parameters of swimming induced by electrical stimulation. Intracellular recordings with potassium-acetate-filled electrodes revealed that the bursts of ventral root discharge induced by NO donors were accompanied by phasic depolarisations in motor neurons. During the inter-burst intervals, periods of substantial membrane hyperpolarization below the normal resting potential were observed, presumably coincident with contralateral ventral root activity. With KCl-filled electrodes, inhibitory potentials were strongly depolarising, suggesting that inhibition was Cl(-)-dependent. The synaptic drive seen in motor neurons after dimming of the illumination was very similar to that induced by the NO donors. NADPH-diaphorase histochemistry identified putative endogenous sources of NO in the central nervous system and the skin. Three populations of bilaterally symmetrical neurons were identified within the brainstem. Some of these neurons had contralateral projections and many had axonal processes that projected to and entered the marginal zones of the spinal cord, suggesting that they were reticulospinal.