Cardiovascular and renal effects of eel and rat atrial natriuretic peptide in rainbow trout,Salmo gairdneri

Summary The renal and in vitro vascular effects of atrial natriuretic peptides have been examined in seveal species of fish. However, comparatively few investigations have described the effects of these peptides on the cardiovascular system in vivo. In the present experiments the dorsal aorta and urinary bladder were cannulated and the effects of atrial natriuretic peptides from rat and eel were monitored in conscious trout during bolus injection or continuous atrial natriuretic peptide infusion. The results show that the initial pressor effect of atrial natriuretic peptides is independent of environmental salinity adaptation (fresh or seawater) and the chemical form of atrial natriuretic peptide injected, but it is affected by the rate of atrial natriuretic peptide administration. This pressor response, and the accompanying diuresis, are mediated through -adrenergic activation. Continuous infusion of either rat or eel atrial natriuretic peptide produces a steady fall in mean arterial blood pressure, which is temporally preceded by an increase in heart rate and a decrease in pulse pressure. Diuresis induced by atrial natriuretic peptides is only partially sustained during continuous infusion. Propranolol partially blocks the increase induced in heart rate by atrial natriuretic peptides, but does not affect either pulse pressure or mean arterial pressure. Propranolol significantly increases urine flow in saline-infused animals but has no apparent effect on animals subjected to infusions of atrial natriuretic peptides. These results indicate that there are multiple foci for the action of atrial natriuretic peptides in trout and that in many instances the effects of atrial natriuretic peptides are mediated through secondary effector systems.Abbreviations ANP atrial natriuretic peptide - bw body weight - PBS phosphate-buffered saline     

Light environments and floristic composition across an open forest-rainforest boundary in northeastern Queensland

Abstract This paper demonstrates the changes in structure, floristics and forest floor light regimes across the boundary between open forest and rainforest at Kirrama in northeastern Queensland. Hemispherical photographs of the canopy were used to estimate spatial and temporal variations in potential (clear-sky) direct and diffuse photosynthetic photon flux density (PPFD = 400–700 nm) across the boundary. Five vegetation zones were identified across the boundary: (A) open forest, (B) tall open forest, (C) tall open forest with a rainforest under-storey, (D) young rainforest, and (E) mature rainforest. During the summer (September–March) direct PPFD declined semi-exponentially across the boundary, while in the winter (April–August), the decline was more linear. However, the decline in diffuse PPFD across the boundary was linear throughout the year. Mean per cent grass cover was positively correlated and mean per cent shrub cover negatively correlated with annual average total PPFD across the boundary. Mean percentages of diffuse radiation relative to that above the forest (diffuse site factors) ranged from 9.8% in the mature rainforest to 66.4% in the open forest, while mean percentages of direct radiation relative to that above the forest (direct site factors) ranged from 2.9–38.3% at the same sites, respectively. Daily PPFD in the rainforest understorey is probably insufficient for the shade intolerant (pioneer) species which prefer the more open environments on the rainforest margin where light intensities and soil temperatures are higher. Towards the rainforest end of zone C, light conditions appear to be similar to those experienced within small treefall gaps in rainforests. Typically, such light conditions are preferred by shade tolerant (primary) tree species that are unable to grow and reproduce successfully in the adjacent shaded understorey and to compete with the fast growing pioneer tree species in the more open end of zone C.     

Quantitative assay for transformation of 3T3 cells by herpes simplex virus type 2.

The interaction of herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) with Swiss/3T3 cells was investigated. Virus-induced cytopathic effects developed in the absence of production of infectious virus. HSV-2 inactivated with UV light (2, 4, 6, and 8 min) also induced cell death in the absence of virus replication. Cell death was not detectable after infection by HSV-2 that had been inactivated by UV irradiation for 10, 12, and 14 min. 3T3 cells infected with UV-inactivated virus (10 and 12 min) continued to replicate past the contact-inhibited monolayer normally associated with these cells. Infection of 3T3 cells with UV-irradiated USV-2 also induced the development of transformed foci. Transformed cells with an epithelioid of fibroblastoid morphology were identified and isolated. All HSV-2-transformed cell lines contained HSV-2-specific antigens detectable by immunofluorescence techniques. The maximum frequency of HSV-2-induced transformation was 3 times 105 PFU per transformed focus, and the observed transformation could be inhibited by pretreatment of the virus with specific antiserum. No type C particles were detected within five cell culture passages after transformation by HSV-2. Type C virus particles were detected after 10 cell culture passages of the HSV-2-transformed cell lines.     

Do positive interactions between marine invaders increase likelihood of invasion into natural and artificial habitats?

Positive species interactions such as facilitation are important for enabling species to persist, especially in stressful conditions, and the nature and strength of facilitation varies along physical and biological gradients. Expansion of coastal infrastructure is creating hotspots of invasive species which can spillover into natural habitats, but the role of positive species interactions associated with biological invasions remains understudied. Theory suggests that stronger biotic pressure in natural habitats inhibits invasion success. In space-limited marine systems, sessile organisms can overcome this limiting resource by settling as an epibiont on a substrate organism – basibiont. Using a series of spatially extensive surveys, we explored the role of invasive and native basibionts in providing habitat for other invasive and native epibionts, and tested whether environmental context (i.e. if the receiving habitat was natural or artificial), altered ecological outcomes. Overall, provision of space by basibionts was more important for invasive epibionts than for native epibionts but was dependent on the environmental context. Invasive basibionts facilitated invasive epibionts in natural habitats, and appeared to be more important for native epibionts in artificial habitats respectively. Native basibionts facilitated invasive, but not native epibionts in both natural and artificial habitats. These results advance our understanding of facilitation and highlight the idiosyncratic nature of biofouling and epibiosis, and the potentially important influence of environmental context. The degree to which native habitat-forming species versus invasive habitat-forming species either do or do not facilitate other native or non-native species is a rich area for investigation. Experimental work is required to disentangle the processes underpinning these patterns.     

Differentiation of the binding of two ligands to a tetrameric protein with a single symmetric active site by 19F NMR

R67 dihydrofolate reductase (R67 DHFR) is a plasmid‐encoded enzyme that confers resistance to the antibacterial drug trimethoprim. R67 DHFR is a tetramer with a single active site that is unusual as both cofactor and substrate are recognized by symmetry‐related residues. Such promiscuity has limited our previous efforts to differentiate ligand binding by NMR. To address this problem, we incorporated fluorine at positions 4, 5, 6, or 7 of the indole rings of tryptophans 38 and 45 and characterized the spectra to determine which probe was optimal for studying ligand binding. Two resonances were observed for all apo proteins. Unexpectedly, the W45 resonance appeared broad, and truncation of the disordered N‐termini resulted in the appearance of one sharp W45 resonance. These results are consistent with interaction of the N‐terminus with W45. Binding of the cofactor broadened W38 for all fluorine probes, whereas substrate, dihydrofolate, binding resulted in the appearance of three new resonances for 4‐ and 5‐fluoroindole labeled protein and severe line broadening for 6‐ and 7‐fluoroindole R67 DHFR. W45 became slightly broader upon ligand binding. With only two peaks in the ¹⁹F NMR spectra, our data were able to differentiate cofactor and substrate binding to the single, symmetric active site of R67 DHFR and yield binding affinities.     

Airway Microbiota in Severe Asthma and Relationship to Asthma Severity and Phenotypes

BackgroundThe lower airways harbor a community of bacterial species which is altered in asthma.ObjectivesWe examined whether the lower airway microbiota were related to measures of asthma severity.MethodsWe prospectively recruited 26 severe asthma, 18 non-severe asthma and 12 healthy subjects. DNA was extracted from induced sputum and PCR amplification of the V3-V5 region of bacterial 16S rRNA gene was performed.ResultsWe obtained 138,218 high quality sequences which were rarefied at 133 sequences/sample. Twenty OTUs had sequences ≥1% of total. There were marked differences in the distribution of Phyla between groups (P = 2.8x10^-118). Bacteroidetes and Fusobacteria were reduced in non-severe and severe asthmatic groups. Proteobacteria were more common in non-severe asthmatics compared to controls (OR = 2.26; 95% CI = 1.94–2.64) and Firmicutes were increased in severe asthmatics compared to controls (OR = 2.15; 95%CI = 1.89–2.45). Streptococcal OTUs amongst the Firmicutes were associated with recent onset asthma, rhinosinusitis and sputum eosinophilia.ConclusionsSputum microbiota in severe asthma differs from healthy controls and non-severe asthmatics, and is characterized by the presence of Streptococcus spp with eosinophilia. Whether these organisms are causative for the pathophysiology of asthma remains to be determined.