Optimal protection of stabilised dry live bacteria from bile toxicity in oral dosage forms by bile acid adsorbent resins

We previously found that dried live bacteria of a vaccine strain can be temporarily sensitive to bile acids and suggested that Bile Adsorbing Resins (BAR) can be used in oral vaccine tablets to protect dried bacteria from intestinal bile. Here, we report a quantitative analysis of the ability of BAR to exclude the dye bromophenol blue from penetrating into matrix tablets and also sections of hard capsule shells. Based on this quantitative analysis, we made a fully optimised formulation, comprising 25% w/w of cholestyramine in Vcaps™ HPMC capsules. This gave effectively 100% protection of viability from 4% bile, with 4200-fold more live bacteria recovered from this formulation compared to unprotected dry bacteria. From the image analysis, we found that the filler material or compaction force used had no measurable effect on dye exclusion but did affect the rate of tablet hydration. Increasing the mass fraction of BAR gave more exclusion of dye up to 25% w/w, after which a plateau was reached and no further dye exclusion was seen. More effective dye exclusion was seen with smaller particle sizes (i.e. cholestyramine) and when the BAR was thoroughly dried and disaggregated. Similar results were found when imaging dye penetration into capsule sections or tablets. The predictions of the dye penetration study were tested using capsules filled with dried attenuated Salmonella vaccine plus different BAR types, and the expected protection from bile was found, validating the imaging study. Surprisingly, depending on the capsule shell material, some protection was given by the capsule alone without adding BAR, with Vcaps™ HPMC capsules providing up to 174-fold protection against 1% bile; faster releasing Vcaps Plus™ HPMC capsules and Coni Snap™ gelatin capsules gave less protection.

Protection of live bacteria from bile acid toxicity using bile acid adsorbing resins

We previously demonstrated that a dry, room temperature stable formulation of a live bacterial vaccine was highly susceptible to bile, and suggested that this will lead to significant loss of viability of any live bacterial formulation released into the intestine using an enteric coating or capsule. We found that bile and acid tolerance is very rapidly recovered after rehydration with buffer or water, raising the possibility that rehydration in the absence of bile prior to release into the intestine might solve the problem of bile toxicity to dried cells. We describe here a novel formulation that combines extensively studied bile acid adsorbent resins with the dried bacteria, to temporarily adsorb bile acids and allow rehydration and recovery of bile resistance of bacteria in the intestine before release. Tablets containing the bile acid adsorbent cholestyramine release 250-fold more live bacteria when dissolved in a bile solution, compared to control tablets without cholestyramine or with a control resin that does not bind bile acids. We propose that a simple enteric coated oral dosage form containing bile acid adsorbent resins will allow improved live bacterial delivery to the intestine via the oral route, a major step towards room temperature stable, easily administered and distributed vaccine pills and other bacterial therapeutics

Incorporation of cis-9,trans-11 or trans-10,cis-12 conjugated linoleic acid into plasma and cellular lipids in healthy men

This study investigated the incorporation of cis-9,trans-11 conjugated linoleic acid (c9,t11 CLA) and trans-10,cis-12-CLA (t10,c12 CLA) into plasma and peripheral blood mononuclear cell (PBMC) lipids when consumed as supplements highly enriched in these isomers. Healthy men (n = 49, age 31 +/- 8 years) consumed one, two, and four capsules containing similar to600 mg of either c9,t11 CIA or t10,c12 CLA per capsule for sequential 8 week periods followed by a 6 week washout before consuming the alternative isomer. Both isomers were incorporated in a dosedependent manner into plasma phosphatidylcholine (PC) (c9,t11 CLA r = 0.779, t10,c12 CLA r = 0.738; P < 0.0001) and cholesteryl ester (CE) (c9,t11 CLA r = 0.706, t10,c12 CLA r = 0.788; P < 0.0001). Only t10,c12 CLA was enriched in plasma nonesterified fatty acids. Both c9,t11 CIA and t10,c12 CLA were incorporated linearly into PBMC total lipids (r = 0.285 and r = 0.273, respectively; P < 0.0005). The highest concentrations of c9,t11 CLA and t10,c12 CLA in PBMC lipids were 3- to 4-fold lower than those in plasma PC and CE. These data suggest that the level of intake is a major determinant of plasma and PBMC CLA content, although PBMCs appear to incorporate both CLA isomers less readily.

Predators of Spotted Flycatcher Muscicapa striata nests in southern England as determined by digital nest cameras

Capsule Avian predators are principally responsible. Aims To document the fate of Spotted Flycatcher nests and to identify the species responsible for nest predation. Methods During 2005-06, purpose-built, remote, digital nest-cameras were deployed at 65 out of 141 Spotted Flycatcher nests monitored in two study areas, one in south Devon and the second on the border of Bedfordshire and Cambridgeshire. Results Of the 141 nests monitored, 90 were successful (non-camera nests, 49 out of 76 successful, camera nests, 41 out of 65). Fate was determined for 63 of the 65 nests monitored by camera, with 20 predation events documented, all of which occurred during daylight hours. Avian predators carried out 17 of the 20 predations, with the principal nest predator identified as Eurasian Jay Garrulus glandarius. The only mammal recorded predating nests was the Domestic Cat Felis catus, the study therefore providing no evidence that Grey Squirrels Sciurus carolinensis are an important predator of Spotted Flycatcher nests. There was no evidence of differences in nest survival rates at nests with and without cameras. Nest remains following predation events gave little clue as to the identity of the predator species responsible. Conclusions Nest-cameras can be useful tools in the identification of nest predators, and may be deployed with no subsequent effect on nest survival. The majority of predation of Spotted Flycatcher nests in this study was by avian predators, principally the Jay. There was little evidence of predation by mammalian predators. Identification of specific nest predators enhances studies of breeding productivity and predation risk.

Ultrastructure of Buddenbrockia allmani n sp (Myxozoa, Malacosporea), a parasite of Lophopus crystallinus (Bryozoa, Phylactolaemata)

Development of a new species of malacosporean myxozoan (Buddenbrockia allmani n. sp.) in the bryozoan Lophopus crystallinus is described. Early stages, represented by isolated cells or small groups, were observed in the host's body wall or body cavity. Multiplication and rearrangement of cells gave an outer cell layer around a central mass. The outer cells made contact by filopodia and established adherens junctions. Sporoplasmosomes were a notable feature of early stages, but these were lost in subsequent development. Typical malacosporean sacs were formed from these groups by attachment of the inner (luminal) cells by a basal lamina to the outer layer (mural cells). Division of luminal cells gave rise to a population of cells that was liberated into the lumen of the sac. Mitotic spindles in open mitosis and prophase stages of meiosis were observed in luminal cells. Centrioles were absent. Detached luminal cells assembled to form spores with four polar capsules and several valve cells surrounding two sporoplasms with secondary cells. Restoration of sporoplasmosomes occurred in primary sporoplasms. A second type of sac was observed with highly irregular mural cells and stellate luminal cells. A radially striated layer and dense granules in the polar capsule wall, and previous data on 18 rDNA sequences enabled assignment of the species to the genus Buddenbrockia, while specific diagnosis relied on the rDNA data and on sac shape and size.

Noncompetitive plasma biokinetics of deuterium-labeled natural and synthetic alpha-tocopherol in healthy men with an apoE4 genotype

Previous studies comparing the biokinetics of deuterated natural (RRR) and synthetic (all-rac) α-tocopherol (vitamin E) used a simultaneous ingestion or competitive uptake approach and reported relative bioavailability ratios close to 2:1, higher than the accepted biopotency ratio of 1.36:1. The aim of the current study was to compare the biokinetics of deuterated natural and synthetic vitamin E using a noncompetitive uptake model both before and after vitamin E supplementation in a distinct population. Healthy men (n = 10) carrying the apolipoprotein (apo)E4 genotype completed a randomized crossover study, comprised of two 4-wk treatments with 400 mg/d (RRR-α-tocopheryl and all-rac-α-tocopheryl acetate) with a 12-wk washout period between treatments. Before and after each treatment period, the subjects consumed a capsule containing 150 mg deuterated α-tocopheryl acetate in either the PRR or all-rac form depending on their treatment regimen. Blood was obtained up to 48 h after ingestion, and tocopherols analyzed by LC/MS. After deuterated all-rac administration, plasma deuterated tocopherol maximum concentrations and area under the concentration vs. time curves (AUC) were lower than those following RRR administration. The RRR:all-rac ratios determined from the deuterated biokinetic profiles (maximum concentration; C-max) and AUCs were 1.35:1 &PLUSMN; 0.17 and 1.33:1 &PLUSMN; 0.18, respectively. The 4-wk supplementation with either PRR or all-rac significantly increased plasma a-tocopherol concentrations (P < 0.001), but decreased the plasma response to newly absorbed deuterated RRR or all-rac α-tocopherol. Using a noncompetitive uptake approach, the relative bioavailability of natural to synthetic vitamin E in apoE4 males was close to the currently accepted biopotency ratio of 1.36:1.

The absorption of vitamin E is influenced by the amount of fat in a meal and the food matrix

Vitamin E absorption requires the presence of fat; however, limited information exists on the influence of fat quantity on optimal absorption. In the present study we compared the absorption of stable-isotope-labelled vitamin E following meals of varying fat content and source. In a randomised four-way cross-over study, eight healthy individuals consumed a capsule containing 150 mg H-2-labelled RRR-alpha-tocopheryl acetate with a test meal of toast with butter (17.5 g fat), cereal with full-fat milk (17.5 g fat), cereal with semi-skimmed milk (2.7 g fat) and water (0g fat). Blood was taken at 0, 0.5, 1, 1.5, 2, 3, 6 and 9 h following ingestion, chylomicrons were isolated, and H-2-labelled alpha-tocopherol was analysed in the chylomicron and plasma samples. There was a significant time (P<0.001) and treatment effect (P<0.001) in H-2-labelled alpha-tocopherol concentration in both chylomicrons and plasma between the test meals. H-2-labelled alpha-tocopherol concentration was significantly greater with the higher-fat toast and butter meal compared with the low-fat cereal meal or water (P< 0.001), and a trend towards greater concentration compared with the high-fat cereal meal (P= 0.065). There was significantly greater H-2-labelled α-tocopherol concentration with the high-fat cereal meal compared with the low-fat cereal meal (P< 0.05). The H-2-labelled alpha-tocopherol concentration following either the low-fat cereal meal or water was low. These results demonstrate that both the amount of fat and the food matrix influence vitamin E absorption. These factors should be considered by consumers and for future vitamin E intervention studies.

Incorporation of cis-9,trans-11 or trans-10,cis-12 conjugated linoleic acid into plasma and cellular lipids in healthy men

This study investigated the incorporation of cis-9,trans-11 conjugated linoleic acid (c9,t11 CLA) and trans-10,cis-12-CLA (t10,c12 CLA) into plasma and peripheral blood mononuclear cell (PBMC) lipids when consumed as supplements highly enriched in these isomers. Healthy men (n = 49, age 31 +/- 8 years) consumed one, two, and four capsules containing similar to600 mg of either c9,t11 CIA or t10,c12 CLA per capsule for sequential 8 week periods followed by a 6 week washout before consuming the alternative isomer. Both isomers were incorporated in a dosedependent manner into plasma phosphatidylcholine (PC) (c9,t11 CLA r = 0.779, t10,c12 CLA r = 0.738; P < 0.0001) and cholesteryl ester (CE) (c9,t11 CLA r = 0.706, t10,c12 CLA r = 0.788; P < 0.0001). Only t10,c12 CLA was enriched in plasma nonesterified fatty acids. Both c9,t11 CIA and t10,c12 CLA were incorporated linearly into PBMC total lipids (r = 0.285 and r = 0.273, respectively; P < 0.0005). The highest concentrations of c9,t11 CLA and t10,c12 CLA in PBMC lipids were 3- to 4-fold lower than those in plasma PC and CE. These data suggest that the level of intake is a major determinant of plasma and PBMC CLA content, although PBMCs appear to incorporate both CLA isomers less readily.

Pupal parasitoid attack influences the relative fitness of Drosophila that have encapsulated larval parasitoids

1. The evolution of host resistance to parasitoid attack will be constrained by two factors: the costs of the ability to defend against attack, and the costs of surviving actual attack. These factors have been investigated using Drosophila melanogaster and its parasitoids as a model system. The costs of defensive ability are expressed as a trade-off with larval competitive ability, whereas the costs of actual defence are exhibited in terms of reduced adult fecundity and size. 2. The costs of actual defence may be ameliorated by the host-choice decisions made by Pachycrepoideus vindemiae, a pupal parasitoid. If larvae that have successfully encapsulated a parasitoid develop into poorer quality hosts, then these may be rejected by ovipositing pupal parasitoids. 3. Pupae developing from larvae that have encapsulated the parasitoid Asobara tabida are smaller and have relatively thinner puparia. Thinner puparia are likely to be associated with a reduction in mechanical strength and possibly with a decrease in desiccation tolerance. 4. Pachycrepoideus vindemiae that develop in capsule-bearing pupae are smaller than those that emerge from previously unattacked hosts. This supports the prediction that ovipositing female P. vindemiae should avoid attacking capsule-bearing hosts. However, in choice experiments with 1-day-old pupae, P. vindemiae females oviposited preferentially in hosts containing a capsule, whereas there was no preference found with 4-day-old hosts. This appears to be a maladaptive host choice decision, as the female pupal parasitoids are preferentially attacking hosts that will result in a reduction of their own fitness. 5. The increased likelihood of attack by a pupal parasitoid is another cost of actual defence against larval parasitoid attack.

The relative fitness of Drosophila melanogaster (Diptera, Drosophilidae) that have successfully defended themselves against the parasitoid Asobara tabida (Hymenoptera, Braconidae)

Drosophila melanogaster larvae defend themselves against parasitoid attack via the process of encapsulation. However, flies that successfully defend them selves have reduced fitness as adults. Adults which carry an encapsulated parasitoid egg are smaller and females produce significantly fewer eggs than controls. Capsule-bearing males allowed repeated copulations with females do not show a reduction in their number of offspring, but those allowed to copulate only once did. No differences were found in time to first oviposition in females, or in time to first copulation in males. We interpret the results as arising from a trade-off between investing resources in factors promoting fecundity and mating success, and in defence against parasitism. The outcome of this investment decision influences the strength of selection for defence against parasitism.

Habitat associations and breeding bird community composition within the city of Bristol, UK

Capsule: Different urban breeding bird communities are associated with different habitat types, but, although community species diversity varies significantly, total bird density does not. Aims: To investigate the association between breeding bird communities and habitats within Bristol, UK and how these communities vary in terms of species diversity and total bird abundance. Methods: Breeding density data for 70 species in the metropolitan area of Bristol, UK were subjected to de-trended correspondence analysis to identify the number of different communities present and their indicator species. These data were then used to identify patterns of habitat association with each community and differences in species richness and total bird density. Results: Three communities were identified: a rural community associated with woodland, managed grassland and inland water; a suburban community associated with buildings and residential gardens; and an intermediate community that shared some of these habitat characteristics. Species richness, but not total bird abundance, was lowest in the suburban community. Conclusion: The diversity of species in urban areas appears to be most dependent upon the availability of patches of natural and semi-natural habitats. Residential gardens support fewer species, but those species that are present may be found at high densities.

Conserved hydrophobic clusters on the surface of the Caf1A usher C-terminal domain are important for F1 antigen assembly

The outer membrane usher protein Caf1A of the plague pathogen Yersinia pestis is responsible for the assembly of a major surface antigen, the F1 capsule. The F1 capsule is mainly formed by thin linear polymers of Caf1 (capsular antigen fraction 1) protein subunits. The Caf1A usher promotes polymerization of subunits and secretion of growing polymers to the cell surface. The usher monomer (811 aa, 90.5 kDa) consists of a large transmembrane β-barrel that forms a secretion channel and three soluble domains. The periplasmic N-terminal domain binds chaperone-subunit complexes supplying new subunits for the growing fiber. The middle domain, which is structurally similar to Caf1 and other fimbrial subunits, serves as a plug that regulates the permeability of the usher. Here we describe the identification, characterization, and crystal structure of the Caf1A usher C-terminal domain (Caf1A(C)). Caf1A(C) is shown to be a periplasmic domain with a seven-stranded β-barrel fold. Analysis of C-terminal truncation mutants of Caf1A demonstrated that the presence of Caf1A(C) is crucial for the function of the usher in vivo, but that it is not required for the initial binding of chaperone-subunit complexes to the usher. Two clusters of conserved hydrophobic residues on the surface of Caf1A(C) were found to be essential for the efficient assembly of surface polymers. These clusters are conserved between the FGL family and the FGS family of chaperone-usher systems.

Svetlana

Svetlana is a series of photographs documenting rehearsals for an opera that was never performed. Written by Waw Pierogi, founder of the 1980s group Xex, little is known of the opera, only that it was inspired by Svetlana, a character from one of their songs and the daughter of Stalin, who defected from the Soviet Union twice. A fictional Svetlana and a bogus Leon Theremin - inventor of the eponymous hands-free electronic musical instrument who was later kidnapped by the KGB - inhabit an archive of photographs from a session of stage rehearsals and location shots. Combining Svetlana’s narrative with a conspiracy to create sound weapons, this documentation of theatre workshops, styled after Bauhaus drama class exercises, produces an entirely spurious story of espionage, sonic weaponry and the clash between love and ideology. The performers sport geometric military costumes, brandishing sculptural forms fashioned after the acoustic locators that preceded radar technology. These redundant locators were still kept in use as props, concealing the introduction of radar from the Germans. They perfectly capture the theatricality of military might and suggest the rhetorical force of sound or even the political power of art. Svetlana was originally produced as part of a residency at S1 Artspace, Sheffield, and was later shown at Tatty Devine, alongside a special capsule collection of jewellery made by Tatty Devine.

Assessing and understanding the impact of stratospheric dynamics and variability on the earth system

Advances in weather and climate research have demonstrated the role of the stratosphere in the Earth system across a wide range of temporal and spatial scales. Stratospheric ozone loss has been identified as a key driver of Southern Hemisphere tropospheric circulation trends, affecting ocean currents and carbon uptake, sea ice, and possibly even the Antarctic ice sheets. Stratospheric variability has also been shown to affect short term and seasonal forecasts, connecting the tropics and midlatitudes and guiding storm track dynamics. The two-way interactions between the stratosphere and the Earth system have motivated the World Climate Research Programme's (WCRP) Stratospheric Processes and Their Role in Climate (SPARC) DynVar activity to investigate the impact of stratospheric dynamics and variability on climate. This assessment will be made possible by two new multi-model datasets. First, roughly 10 models with a well resolved stratosphere are participating in the Coupled Model Intercomparison Project 5 (CMIP5), providing the first multi-model ensemble of climate simulations coupled from the stratopause to the sea floor. Second, the Stratosphere Historical Forecasting Project (SHFP) of WCRP's Climate Variability and predictability (CLIVAR) program is forming a multi-model set of seasonal hindcasts with stratosphere resolving models, revealing the impact of both stratospheric initial conditions and dynamics on intraseasonal prediction. The CMIP5 and SHFP model-data sets will offer an unprecedented opportunity to understand the role of the stratosphere in the natural and forced variability of the Earth system and to determine whether incorporating knowledge of the middle atmosphere improves seasonal forecasts and climate projections. Capsule New modeling efforts will provide unprecedented opportunities to harness our knowledge of the stratosphere to improve weather and climate prediction.