In vitro culture of the strobilar stage of Echinococcus granulosus from protoscoleces of human, camel, cattle, sheep and goat origin from Kenya and buffalo origin from India

Protoscoleces from human, camel, cattle, sheep, goat (all from Kenya) and buffalo (from India) hydatid cysts were cultured under identical conditions in vitro using the diphasic culture system of Smyth (1979b). Organisms from all sources grew and segmented in culture. Genital anlagen developed in all cultured worms but further genital differentiation occurred only in cultures of cattle (testes) and camel (testes and genital pore) material. The possible significance of these results is discussed in relation to the general epidemiology of hydatid disease and the potential infectivity of the different strains to man.     

Canine reproduction: I. Reaction of animals to collection of semen and insemination procedures

Two methods of semen collection were attempted on ten untrained dogs of several breeds. Manual manipulation as described proved most reliable for untrained dogs. Electro-ejaculation was attempted unsucessfully with twelve additional dogs.Two methods of insemination were compared. Equipment found most satisfactory is described and a modification suggested.     

Monolayer properties of chloroplast lipids

The properties of seven monogalactosyldiacylglycerols and six digalactosyldiacylglycerols, isolated from photosynthetic membranes and possessing different levels of fatty acid unsaturation, have been studied by the monolayer technique and compared with those of the fully saturated compounds. In addition, the monolayer properties of sulphoquinovosyldiacylglycerols and phosphatidylglycerols from higher plant chloroplasts, and several hexadecenoic acids have been measured.Monogalactosyldiacylglycerols containing saturated fatty acids form a condensed monolayer similar to that of saturated phosphatidylcholines. The naturally occurring monogalactosyldiacylglycerols, of which the double bond index ranged from 0.6 to 3.9, possessed comparable force-area curves suggesting that headgroup interactions play a more important role in packing behaviour than in phosphatidylcholines. Although digalactosyldiacylglycerols containing fully saturated fatty acids form a more expanded monolayer than the corresponding monogalactosyldiacylglycerols, the degree of expansion of the monolayer due to the presence of unsaturated fatty acids in the naturally occurring digalactosyldiacylglycerols is much less than in monogalactosyldiacylglycerols. Monogalactosyldiacylglycerols and digalactosyldiacylglycerols from a single species have very similar monolayer properties, and the presence of sulphoquinovosyldiacylglycerols and phosphatidylglycerols in the proportions in which they occur in higher plant chloroplasts does not have any condensing effect on a monolayer of galactolipids     

Helping decision making for reliable and cost‐effective 2b‐RAD sequencing and genotyping analyses in non‐model species

High‐throughput sequencing has revolutionized population and conservation genetics. RAD sequencing methods, such as 2b‐RAD, can be used on species lacking a reference genome. However, transferring protocols across taxa can potentially lead to poor results. We tested two different IIB enzymes (AlfI and CspCI) on two species with different genome sizes (the loggerhead turtle Caretta caretta and the sharpsnout seabream Diplodus puntazzo) to build a set of guidelines to improve 2b‐RAD protocols on non‐model organisms while optimising costs. Good results were obtained even with degraded samples, showing the value of 2b‐RAD in studies with poor DNA quality. However, library quality was found to be a critical parameter on the number of reads and loci obtained for genotyping. Resampling analyses with different number of reads per individual showed a trade‐off between number of loci and number of reads per sample. The resulting accumulation curves can be used as a tool to calculate the number of sequences per individual needed to reach a mean depth ≥20 reads to acquire good genotyping results. Finally, we demonstrated that selective‐base ligation does not affect genomic differentiation between individuals, indicating that this technique can be used in species with large genome sizes to adjust the number of loci to the study scope, to reduce sequencing costs and to maintain suitable sequencing depth for a reliable genotyping without compromising the results. Here, we provide a set of guidelines to improve 2b‐RAD protocols on non‐model organisms with different genome sizes, helping decision‐making for a reliable and cost‐effective genotyping.     

Metabolipidomic profiling reveals an age‐related deficiency of skeletal muscle pro‐resolving mediators that contributes to maladaptive tissue remodeling

Specialized pro‐resolving mediators actively limit inflammation and support tissue regeneration, but their role in age‐related muscle dysfunction has not been explored. We profiled the mediator lipidome of aging muscle via liquid chromatography‐tandem mass spectrometry and tested whether treatment with the pro‐resolving mediator resolvin D1 (RvD1) could rejuvenate the regenerative ability of aged muscle. Aged mice displayed chronic muscle inflammation and this was associated with a basal deficiency of pro‐resolving mediators 8‐oxo‐RvD1, resolvin E3, and maresin 1, as well as many anti‐inflammatory cytochrome P450‐derived lipid epoxides. Following muscle injury, young and aged mice produced similar amounts of most pro‐inflammatory eicosanoid metabolites of cyclooxygenase (e.g., prostaglandin E₂) and 12‐lipoxygenase (e.g., 12‐hydroxy‐eicosatetraenoic acid), but aged mice produced fewer markers of pro‐resolving mediators including the lipoxins (15‐hydroxy‐eicosatetraenoic acid), D‐resolvins/protectins (17‐hydroxy‐docosahexaenoic acid), E‐resolvins (18‐hydroxy‐eicosapentaenoic acid), and maresins (14‐hydroxy‐docosahexaenoic acid). Similar absences of downstream pro‐resolving mediators including lipoxin A₄, resolvin D6, protectin D1/DX, and maresin 1 in aged muscle were associated with greater inflammation, impaired myofiber regeneration, and delayed recovery of strength. Daily intraperitoneal injection of RvD1 had minimal impact on intramuscular leukocyte infiltration and myofiber regeneration but suppressed inflammatory cytokine expression, limited fibrosis, and improved recovery of muscle function. We conclude that aging results in deficient local biosynthesis of specialized pro‐resolving mediators in muscle and that immunoresolvents may be attractive novel therapeutics for the treatment of muscular injuries and associated pain in the elderly, due to positive effects on recovery of muscle function without the negative side effects on tissue regeneration of non‐steroidal anti‐inflammatory drugs.     

Controllers for imposing continuum-to-molecular boundary conditions in arbitrary fluid flow geometries

We present a new parallelised controller for steering an arbitrary geometric region of a molecular dynamics (MD) simulation towards a desired thermodynamic and hydrodynamic state. We show that the controllers may be applied anywhere in the domain to set accurately an initial MD state, or solely at boundary regions to prescribe non-periodic boundary conditions (PBCs) in MD simulations. The mean molecular structure and velocity autocorrelation function remain unchanged (when sampled a few molecular diameters away from the constrained region) when compared with those distributions measured using PBCs. To demonstrate the capability of our new controllers, we apply them as non-PBCs in parallel to a complex MD mixing nano-channel and in a hybrid MD continuum simulation with a complex coupling region. The controller methodology is easily extendable to polyatomic MD fluids.     

Generation of initial molecular dynamics configurations in arbitrary geometries and in parallel

A computational pre-processing tool for generating initial configurations of molecules for molecular dynamics simulations in geometries described by a mesh of unstructured arbitrary polyhedra is described. The mesh is divided into separate zones and each can be filled with a single crystal lattice of atoms. Each zone is filled by creating an expanding cube of crystal unit cells, initiated from an anchor point for the lattice. Each unit cell places the appropriate atoms for the user-specified crystal structure and orientation. The cube expands until the entire zone is filled with the lattice; zones with concave and disconnected volumes may be filled. When the mesh is spatially decomposed into portions for distributed parallel processing, each portion may be filled independently, meaning that the entire molecular system never needs to fit onto a single processor, allowing very large systems to be created. The computational time required to fill a zone with molecules scales linearly with the number of cells in the zone for a fixed number of molecules, and better than linearly with the number of molecules for a fixed number of mesh cells. Our tool, molConfig, has been implemented in the open source C++ code OpenFOAM.