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Biomedical subjects

S M Rea

Publications and source records attributed to S M Rea.

8 recordsLinked to original sources

Reducing methane emissions in sheep by immunization against rumen methanogens.

This work was conducted to determine if methane emissions from sheep immunized with an anti-methanogen vaccine were significantly lower than methane emissions from non-immunized sheep, to test the effectiveness of two different vaccine formulations (VF) on methane abatement, and to compare methane emissions measured using a closed-circuit respiration chamber and the sulphur-hexafluoride (SF6) tracer technique. Thirty mature wether sheep were randomly allocated to three treatment groups (n = 10). One group received an immunization of adjuvant only on days 0 and 153 (control), a second group received an immunization with a 3-methanogen mix on days 0 and 153 (VF3 + 3), and a third group received an immunization of a 7-methanogen mix on day 0 followed by a 3-methanogen mix on day 153 (VF7 + 3). Four weeks post-secondary immunization, there was a significant 7.7% reduction in methane production per kg dry matter intake in the VF7 + 3 group compared to the controls (P = 0.051). However, methane emissions from sheep immunized with VF7 + 3 were not significantly different when compared to the sheep in the control group (P = 0.883). The average IgG and IgA antibody titres in both plasma and saliva of the VF3 + 3 immunized sheep were four to nine times higher than those immunized with VF7 + 3 (P< 0.001) at both 3 and 6 weeks post-secondary immunization. Data also revealed that SF6 methane estimates were consistently higher than the respiration chamber estimates and that there was no significant correlation between the SF6 methane estimates and the respiration chamber methane estimates (R2 = 0.11).

Adjuvants, Immunologic↗

Proliferation and differentiation of osteoblast-like cells on apatite-wollastonite/polyethylene composites.

Glass-ceramic apatite-wollastonite (A-W)/high-density polyethylene composite (AWPEX) materials have been designed to match the mechanical strength of human cortical bone and to provide favourable bioactivity, with potential use in many orthopaedic applications. To better understand AWPEX properties, the effects of surface finish and ceramic filler size and content on osteoblast-like cell attachment, proliferation, and differentiation were examined. Glass-ceramic content was tested at 30 and 50 vol% and median particle size at 4.5 and 7.7 microm. Samples were prepared as 1 x 10 x 10 mm(3) tiles with polished or rough surfaces, sterilized by gamma irradiation (2.5 Mrad), and characterized by scanning electron microscopy (SEM) and surface profilometry. Saos-2 human osteoblast-like cells were cultured on each surface at an initial concentration of 4500 cells/cm(2) for 1, 3, or 7 days. At each time point, adenosine triphosphate and alkaline phosphatase levels were measured to assess cell number and osteoblast differentiation. SEM imaging of cells on the composite surfaces showed preferential cell attachment to filler particles within the polymer matrix. Significant biochemical assay differences were found at 7 days, confirmed by ANOVA post-hoc testing using Bonferroni's correction. Overall, increased exposure of the glass-ceramic A-W phase in AWPEX through surface polishing, higher volume fraction and/or larger particle size was found to lead to an improved cell response.

Apatites↗

Bioactivity of ceramic-polymer composites with varied composition and surface topography.

HAPEX trade mark (40 vol % hydroxyapatite in a high-density polyethylene matrix) and AWPEX (40 vol % glass-ceramic apatite-wollastonite in a high-density polyethylene matrix) are composites designed to provide bioactivity and to match the mechanical properties of human cortical bone. HAPEX trade mark has had clinical success in middle ear and orbital implants, and there is great potential for further orthopaedic applications of these materials. However, more detailed in vitro investigations must be performed to better understand the biological interactions of the composites. In this study, the bioactivity of each material was assessed. Specifically, the effects of controlled surface topography and ceramic filler composition on apatite layer formation in acellular simulated body fluid (SBF) with ion concentration similar to those of human blood plasma were examined. Samples were prepared as 1 x 10 x 10 mm(3) tiles with polished, roughened or parallel-grooved surface finishes, and were incubated in 20 ml of SBF at 36.5 degrees C for one, three, seven or 14 days. The formation of an apatite layer on the composite surface after immersion was demonstrated by thin-film X-ray diffraction, environmental scanning electron microscopy and energy dispersive X-ray analysis. Variations in sample weight and solution pH over the period of incubation were also recorded. Significant differences were found between the two materials tested, with greater bioactivity in AWPEX than HAPEX trade mark. Results also showed surface topography to be important, with rougher samples correlated to earlier apatite formation. Osteoblast-like cells proliferated favourably on both composite materials, with many filopodia connections, preferential attachment to ceramic particles and contact guidance effects evident.

Apatites↗

The microbial composition of three limnologically disparate hypersaline Antarctic lakes.

16S rRNA clone library analysis was used to examine the biodiversity and community structure within the sediments of three hypersaline Antarctic lakes. Compared to sediment of low to moderate salinity Antarctic lakes the species richness of the hypersaline lake sediments was 2-20 times lower. The community of Deep Lake (32% salinity, average sediment temperature -15 degrees C) was made up almost entirely of halophilic Archaea. The sediment communities of two meromictic hypersaline lakes, Organic Lake (20% salinity, -7 degrees C) and Ekho Lake (15% salinity, 15 degrees C) were more complex, containing phylotypes clustering within the Proteobacteria and Cytophagales divisions and with algal chloroplasts. Many phylotypes of these lakes were related to taxa more adapted to marine-like salinity and perhaps derive from bacteria exported into the sediment from the lower salinity surface waters. The Ekho Lake clone library contained several major phylotypes related to the Haloanaerobiales, the growth of which appears to be promoted by the comparatively high in situ temperature of this lake.

Antarctic Regions↗

Diversity and community structure within anoxic sediment from marine salinity meromictic lakes and a coastal meromictic marine basin, Vestfold Hilds, Eastern Antarctica.

16S rDNA clone library analysis was used to examine the biodiversity and community structure within anoxic sediments of several marine-type salinity meromictic lakes and a coastal marine basin located in the Vestfolds Hills area of Eastern Antarctica. From 69 to 130 (555 total) 16S rDNA clones were analysed from each sediment sample, and restriction fragment length polymorphism (RFLP) and sequence analysis grouped the clones into 202 distinct phylotypes (a clone group with sequence similarity of >0.98). A number of phylotypes and phylotype groups predominated in all libraries, with a group of 10 phylotypes (31% of clones) forming a novel deep branch within the low G+C Gram-positive division. Other abundant phylotypes detected in several different clone libraries grouped with Prochlorococcus cyanobacteria, diatom chloroplasts, delta proteobacteria (Desulfosarcina group, Syntrophus and Geobacterl Pelobacter/Desulphuromonas group), order Chlamydiales (Parachlamydiaceae) and Spirochaetales (wall-less Antarctic spirochaetes). Most archaeal clones detected (3.1% of clones) belonged to a highly diverged group of Euryarchaeota clustering with clones previously detected in rice soil, aquifer sediments and hydrothermal vent material. Little similarity existed between the phylotypes detected in this study and other clone libraries based on marine sediment, suggesting that an enormous prokaryotic diversity occurs within marine and marine-derived sediments.

Antarctic Regions↗

Shewanella gelidimarina sp. nov. and Shewanella frigidimarina sp. nov., novel Antarctic species with the ability to produce eicosapentaenoic acid (20:5 omega 3) and grow anaerobically by dissimilatory Fe(III) reduction.

A polyphasic taxonomic study was performed to characterize dissimilatory iron-reducing strains mostly isolated from Antarctic sea ice. The strains were isolated from samples of congelated (land-fast) sea ice, grease ice, and ice algal biomass collected from the coastal areas of the Vestfold Hills in eastern Antarctica (68 degrees S 78 degrees E). The strains were facultatively anaerobic, motile, and rod shaped, were capable of anaerobic growth either by fermentation of carbohydrates or by anaerobic respiration, and utilized a variety of electron acceptors, including nitrate, ferric compounds, and trimethylamine N-oxide. A phylogenetic analysis performed with 16S rRNA sequences showed that the isolates formed two groups representing novel lineages in the genus Shewanella. The first novel group included seawater-requiring, psychrophilic, chitinolytic strains which had DNA G + C contents of 48 mol%. The members of the second strain group were psychrotrophic and did not require seawater but could tolerate up to 9% NaCl. The strains of this group were also unable to degrade polysaccharides but could utilize a number of monosaccharides and disaccharides and had G + C contents of 40 to 43 mol%. The whole-cell-derived fatty acid profiles of the sea ice isolates were found to be similar to the profiles obtained for other Shewanella species. The omega-3 polyunsaturated fatty acid eicosapentaenoic acid (EPA) (20:5 omega 3) was detected in all of the sea ice isolates at levels ranging from 2 to 16% of the total fatty acids. EPA was also found at high levels in Shewanella hanedai (19 to 22%) and Shewanella benthica (16 to 18%) but was absent in Shewanella alga and Shewanella putrefaciens. On the basis of polyphasic taxonomic data, the Antarctic iron-reducing strains are placed in two new species, Shewanella frigidimarina sp. nov. (type strain, ACAM 591) and Shewanella gelidimarina sp. nov. (type strain, ACAM 456).

Antarctic Regions↗

Electron microscopy of mammalian cells in the absence of fixing, freezing, dehydration, or specimen coating.

Human osteoblast-like (bone-forming) cells were imaged using environmental scanning electron microscopy (ESEM). The cells were hydrated, unfrozen, and uncoated. Specimens were cooled to 3 degrees C and imaged in water vapor, with partial pressures varying from saturated conditions to a humidity of approximately 50%, relative to pure water. The ESEM images show the presence of cell nuclei, nucleoli, and cytoplasmic membranes. Comparisons between chemically fixed and unfixed specimens (neither dried nor coated) show that cell morphologies are similar in both cases. These results are compared with a fixed, dried, carbon-coated specimen. Thermodynamic and kinetic arguments are used to show that humidities significantly lower than 100% correspond to metastable states suitable for stabilizing hydrated biological tissues and cells. The ability to perform observations with minimal specimen preparation is potentially useful for studying interactions between mammalian cells and biomaterials that are developed for tissue engineering. The methods employed are equally applicable to the study of specimens in the biological, materials, and physical sciences where careful control over specimen stability is required.

Cytological Techniques↗