Metabolic enzyme activities in black bream (Acanthopagrus butcheri) from the Swan-Canning Estuary, Western Australia

The Swan-Canning estuary, in southwestern Australia, is subject to frequent algal blooms and associated periods of hypoxia due to high levels of nutrients in stormwater runoff and sewage spills. Fish in which cellular respiration is impaired due to chronic exposure to non-nutrient pollutants in the water will have a reduced ability to survive these periods of high stress. In order to investigate if metabolic respiration in black bream (Acanthopagrus butcheri) was altered, fish were collected from five sites in the Swan-Canning estuary in summer 2001, summer 2002 and winter 2002. Aerobic and anaerobic capacities were estimated by measuring the enzymes cytochrome C oxidase (CCO) and lactate dehydrogenase (LDH). Neither seasonal or annual trends, nor upstream or downstream gradients were observed in either biomarker. The fish collected from the Barrack Street site, which is close to the Perth Central Business District, were heavily challenged in their aerobic capacity in the summer months compared to the other sites. In addition, the fish at Barrack Street displayed an altered anaerobic capacity. It is likely that the impaired metabolic capacity of the fish at Barrack Street reduces the fishes' ability to survive the frequent algal blooms within the estuary.     

Effect of alemtuzumab-based T-cell depletion on graft compositional change in vitro and immune reconstitution early after allogeneic stem cell transplantation

Background aimsTo reduce the risk of graft-versus-host disease (GVHD) after allogeneic stem cell transplantation (alloSCT), T-cell depletion (TCD) of grafts can be performed by the addition of alemtuzumab (ALT) “to the bag” (in vitro) before transplantation. In this prospective study, the authors analyzed the effect of in vitro incubation with 20 mg ALT on the composition of grafts prior to graft infusion. Furthermore, the authors assessed whether graft composition at the moment of infusion was predictive for T-cell reconstitution and development of GVHD early after TCD alloSCT.MethodsSixty granulocyte colony-stimulating factor-mobilized stem cell grafts were obtained from ≥9/10 HLA-matched related and unrelated donors. The composition of the grafts was analyzed by flow cytometry before and after in vitro incubation with ALT. T-cell reconstitution and incidence of severe GVHD were monitored until 12 weeks after transplantation.ResultsIn vitro incubation of grafts with 20 mg ALT resulted in an initial median depletion efficiency of T-cell receptor (TCR) α/β T cells of 96.7% (range, 63.5–99.8%), followed by subsequent depletion in vivo. Graft volumes and absolute leukocyte counts of grafts before the addition of ALT were not predictive for the efficiency of TCR α/β T-cell depletion. CD4^pos T cells were depleted more efficiently than CD8^pos T cells, and naive and regulatory T cells were depleted more efficiently than memory and effector T cells. This differential depletion of T-cell subsets was in line with their reported differential CD52 expression. In vitro depletion efficiencies and absolute numbers of (naive) TCR α/β T cells in the grafts after ALT incubation were not predictive for T-cell reconstitution or development of GVHD post- alloSCT.ConclusionsThe addition of ALT to the bag is an easy, fast and generally applicable strategy to prevent GVHD in patients receiving alloSCT after myeloablative or non-myeloablative conditioning because of the efficient differential depletion of donor-derived lymphocytes and T cells.     

Temporal variations and pond age effect on plankton communities in semi-intensive freshwater marron (Cherax cainii,Austin and Ryan, 2002) earthen aquaculture ponds in Western Australia

The abundance and diversity of the plankton community represents the health of the aquatic ecosystem, and plays an important role in the growth of cultured animals under aquaculture conditions. The temporal variations of plankton abundance, taxonomic composition, diversity, evenness and species richness were studied in three old and three new semi-intensive marron (Cherax cainii, Austin and Ryan, 2002) ponds. Water parameters such as temperature, dissolved oxygen, pH, turbidity, TAN, nitrite, nitrate and reactive phosphate were recorded, and plankton samples were collected every two months, for one year of juvenile production cycle. A total of twenty-six phytoplankton and seven zooplankton genera were recorded. Chlorophyceae was the dominant class of phytoplankton throughout the year, followed by Trebouxiophyceae. Rotifera comprised 49.8% of the total zooplankton community (individuals L^?1), the largest proportion of any group. Temporal variations impacted the plankton abundance and community structure, and plankton abundance were more abundant during summer. The pond age did not influence the phytoplankton abundance, whereas zooplankton abundance was higher in older ponds.     

Tolerance and Physiological Correlates of Neuromuscular Electrical Stimulation in COPD: A Pilot Study

Rationale

Neuromuscular electrical stimulation (NMES) of the lower limbs is an emerging training strategy in patients with COPD. The efficacy of this technique is related to the intensity of the stimulation that is applied during the training sessions. However, little is known about tolerance to stimulation current intensity and physiological factors that could determine it. Our goal was to find potential physiological predictors of the tolerance to increasing NMES stimulation intensity in patients with mild to severe COPD.

Methods

20 patients with COPD (FEV₁ = 54±14% pred.) completed 2 supervised NMES sessions followed by 5 self-directed sessions at home and one final supervised session. NMES was applied simultaneously to both quadriceps for 45 minutes, at a stimulation frequency of 50 Hz. Spirometry, body composition, muscle function and aerobic capacity were assessed at baseline. Cardiorespiratory responses, leg discomfort, muscle fatigue and markers of systemic inflammation were assessed during or after the last NMES session. Tolerance to NMES was quantified as the increase in current intensity from the initial to the final NMES session (ΔInt).

Results

Mean ΔInt was 12±10 mA. FEV₁, fat-free-mass, quadriceps strength, aerobic capacity and leg discomfort during the last NMES session positively correlated with ΔInt (r = 0.42 to 0.64, all p≤0.06) while post/pre NMES IL-6 ratio negatively correlated with ΔInt (r = −0.57, p = 0.001). FEV₁, leg discomfort during last NMES session and post/pre IL-6 ratio to NMES were independent factors of variance in ΔInt (r² = 0.72, p = 0.001).

Conclusion

Lower tolerance to NMES was associated with increasing airflow obstruction, low tolerance to leg discomfort during NMES and the magnitude of the IL-6 response after NMES.

Trial Registration

ClinicalTrials.gov {"type":"clinical-trial","attrs":{"text":"NCT00809120","term_id":"NCT00809120"}}NCT00809120     

The SciZ protein anchors the enteroaggregative Escherichia coli Type VI secretion system to the cell wall

Type VI secretion systems (T6SS) are multi‐component machines encoded within the genomes of most Gram‐negative bacteria that associate with plant, animal and/or human cells, and therefore are considered as potential virulence factors. We recently launched a study on the Sci‐1 T6SS of enteroaggregative Escherichia coli (EAEC). The Sci‐1 T6SS is composed of all or a subset of the 21 gene products encoded within the cluster, 13 of which are shared by all T6SS identified so far. In the present work, we focussed our attention on the SciZ protein. We first showed that SciZ is required for the release of the Hcp protein in the culture supernatant and for efficient biofilm formation, demonstrating that SciZ is necessary for EAEC T6SS function. Indeed, SciZ forms a complex with SciP, SciS and SciN, three core components of the transport apparatus. Fractionation and topology studies showed that SciZ is a polytopic inner membrane protein with three trans‐membrane segments. Computer analyses identified a motif shared by peptidoglycan binding proteins of the OmpA family in the SciZ periplasmic domain. Using in vivo and in vitro binding assays, we showed that this motif anchors the SciZ protein to the cell wall and is required for T6SS function.     

Biomarkers of exposure in fish inhabiting the Swan–Canning Estuary, Western Australia – a preliminary study

The estuarine portion of the Swan–Canning riversystem runs through the centre of Perth,Western Australia's capital city, with apopulation of approximately 1.4 million people. Little is known about impact of chemicalsentering the estuary via road runoff andstormwater drains on biota inhabiting thesystem. Black bream (Acanthopagrusbutcheri) were collected from seven sites inthe Swan–Canning estuary during August andSeptember 2000, at the end of the winter (wet)season. Serum sorbitol dehydrogenase (s-SDH)was unaffected by the sex of the fish and nosignificant differences were observed betweenthe sites indicating that the measuredethoxyresorufin-O-deethylase (EROD)activity was not hindered by hepatic tissuedamage. The black bream were in an advancedstage of gonad maturation, which affected ERODhepatic activity with lower EROD activity infemale compared to male fish. EROD activityand bile metabolite levels were significantlyhigher at the site closest to the Perth CentralBusiness District, while most downstream sitewas the least impacted, which may be due totidal flushing of the lower estuary by marinewaters. The ratio of naphthalene-type tobenzo(a) pyrene (B(a)P)-typemetabolites suggests that the source ofpetroleum hydrocarbons within the river systemis a mixture polycyclic aromatic hydrocarbons(PAHs) from pyrolytic origin and from unburntfuels. Biomarker levels in the black breamindicate that major roads and drains aresignificant contributors of mixed functionoxygenase (MFO) inducing chemicals includingpolycyclic aromatic hydrocarbons (PAH) into theSwan–Canning estuary and that there is noupstream or downstream gradient in biomarkerresponse.     

Development of LBP110 expression by neural crest-derived enteric precursors: Migration and differentiation potential in ls/ls mutant mice

Neural crest-derived cells acquire a 110-kD laminin-binding protein (LBP110) when they colonize the murine bowel. Laminin stimulates LBP110-expressing cells to develop as neurons. We have followed the development of LBP110 by neural crest-derived cells as they enter the gut of control and ls/ls mutant mice. The expression of neurofilament and choline acetyltransferase was used as markers of a neuronal phenotype. Tyrosine hydroxylase was used as a marker for the mash-1-dependent lineage of enteric precursors, while calcitonin gene-related peptide was used as a marker for the mash-1-independent lineage of crest-derived cells. A subset of cells expressing LBP110 was located along the vagi at E10 at cervical and thoracic levels. At E12, cells expressing LBP110 extended from the foregut to the midgut. The expression of neurofilament protein lagged behind that of LBP110 by about 0.5 day and then became coincident with LBP110 immunoreactivity. By E15, cells doubly labeled with antibodies to LBP110 and neurofilament protein were located along the entire extent of the bowel up to but not including the terminal colon. By E16, both the proximal and terminal colon contained cells expressing LBP110 and neurofilaments. The pattern of immunoreactivity could not be distinguished between ls/ls and control animals prior to E16. By E16, when the terminal colon of control animals contained many cells expressing LBP110 and neurofilaments, the terminal colon of ls/ls animals lacked cells expressing these proteins; nevertheless, structures outside of the terminal colon were heavily endowed with cells expressing LBP110 and neurofilaments. These ectopically located cells derived from both mash-1-dependent and -independent lineages of crest-derived precursors. © 1998 John Wiley & Sons, Inc. J Neurobiol 35: 341–354, 1998     

Escherichia coli tol-pal Mutants Form Outer Membrane Vesicles

Mutations in the tol-pal genes induce pleiotropic effects such as release of periplasmic proteins into the extracellular medium and hypersensitivity to drugs and detergents. Other outer membrane defective strains such as tolC, lpp, and rfa mutations are also altered in their outer membrane permeability. In this study, electron microscopy and Western blot analyses were used to show that strains with mutations in each of the tol-pal genes formed outer membrane vesicles after growth in standard liquid or solid media. This phenotype was not observed in tolC and rfaD cells in the same conditions. A tolA deletion in three different Escherichia coli strains was shown to lead to elevated amounts of vesicles. These results, together with plasmid complementation experiments, indicated that the formation of vesicles resulted from the defect of any of the Tol-Pal proteins. The vesicles contained outer membrane trimeric porins correctly exposed at the cell surface. Pal outer membrane lipoprotein was also immunodetected in the vesicle fraction of tol strains. The results are discussed in view of the role of the Tol-Pal transenvelope proteins in maintaining outer membrane integrity by contributing to target or integrate newly synthesized components of this structure.