Liquid storage of Ostrich (Struthio camelus) semen at 5 °C through intermediate dilution.

Dilution rate, dilution temperature and storage time have been recognized as vital steps in the processing of semen for storage before artificial insemination. The objective of this study was to determine optimal dilution and dilution temperature with an ostrich-specific semen extender for chilled storage. Four preselected ostrich (Struthio camelus var. domesticus) males, known for their ease of collection and specific semen quality parameters, were collected using the "dummy" female method. Dilution of 384 semen samples, at rates of 1:1, 1:2, 1:4 and 1:8 semen/diluent ratio with a diluent set at 5, 21 and 38 °C was performed and stored for 48 h at 5 °C. In vitro sperm function tests were conducted to evaluate treated semen during different storage intervals of 1, 5, 24 and 48 h. Motility and kinematic parameters were measured by the Sperm Class Analyzer®, the percentage live sperm measured by fluorescence SYBR14®/PI (LIVE/DEAD®), the percentage of sperm able to resist the hypo-osmotic swelling (HOS) stress test and sperm morphology determined by Nigrosin-Eosin staining. Progressive motility (PMOT), motility (MOT), sperm kinematics, LIVE and HOS were best (P < 0.05) maintained at a higher dilution of 1:4-1:8. The beneficial effect (P < 0.05) of a higher dilution temperature (21 °C) was prominent in terms of PMOT at a higher dilution. Storage of chilled semen at 5 °C requires dilution, at interpolated rates of 1:6-1:7, together with an extender temperature of 21 °C, to maintain optimal sperm function with minimal loss over a 48 h storage period.

Understanding the crystallographic and nanomechanical properties of bryozoans.

This study examines how microscale differences in skeletal ultrastructure affect the crystallographic and nanomechanical properties of two related bryozoan species: (i) Hornera currieae, which is found at relatively quiescent depths of c. 1000 m, and (ii) Hornera robusta, which lives at depths of 50-400 m where it is exposed to currents and storm waves. Microstructural and Electron Backscatter Diffraction (EBSD) observations show that in both species the secondary walls are composed of low-Mg calcite crystallites that grow with their c-axes perpendicular to the wall. Branches in H. currieae develop a strong preferred orientation of the calcite c-axes, while in H. robusta the c-axes are more scattered. Microstructural observations suggest that the degree of scattering is controlled by the underlying morphology of the skeletons: in H. currieae the laminated branch walls are smooth and relatively uninterrupted, whereas the wall architecture of H. robusta is modified by numerous deflections, forming pustules and ridges associated with microscopic tubules. Modelling of the Young's modulus and measurements of nanoindentation hardness indicate that the observed scattering of the crystallite c-axes affects the elastic modulus and nanohardness of the branches, and therefore controls the mechanical properties of the skeletal walls. At relatively high pressure in deep waters, the anisotropic skeletal architecture of H. currieae is aimed at concentrating elasticity normal to the skeleton wall. In comparison, in the relatively shallow and active hydrographic regime of the continental shelf, the elastically isotropic skeleton of H. robusta is designed to increase protection from external predators and stronger omni-directional currents.

Strong, Non-specific Adhesion Using C-Lectin Heterotrimers in a Molluscan Defensive Secretion.

The defensive mucus produced by the slug Arion subfuscus is tough and remarkably sticky. It spreads rapidly and adheres strongly to a wide range of surfaces. The adhesion is equally strong on wettable (glass) and non-wettable (plastic) surfaces. The adhesion appears to depend on a group of proteins that adsorb equally well to a wide range of different natural and artificial surfaces. Prominent among these proteins were those that distinguish the adhesive secretion from the non-adhesive mucus. The adhesive proteins were not washed off by non-ionic detergent. Similarly, the adhesion of the glue as a whole was not affected by this treatment. In contrast, high salt concentrations washed the most abundant adhesive proteins off the surfaces, and correspondingly weakened the glue's attachment. The most abundant of the adhesive proteins were C-lectins, which appear to form heterotrimers. These and other lectin-like proteins in slug glue have a high proportion of aromatic amino acids at conserved locations, and are relatively small and often basic. The aromatic and cationic side chains may provide a powerful combination promoting and maintaining surface adhesion.

Effects of maternal obesity in an ovine model on metabolic outcomes in F2 adults and F3 neonates.

Accumulating evidence suggests that indications of metabolic syndrome can be inherited through the germline as a result of maternal obesity. We hypothesized that diet-induced maternal obesity during gestation would program metabolic consequences for multiple generations of offspring, even when first, second, and third generation offspring (F1, F2, F3, respectively) were fed only to requirements. Control (CON) and obese (OB) ewes (generation 0; F0) were bred to a single ram to produce the first generation of offspring (F1). From 60 d prior to conception through term, CONF0 ate 100% National Research Council recommendations (NRC), while OBF0 ewes ate 150% NRC. All F1, F2, and F3 ate 100% NRC after weaning. All mature F1 ewes were bred to a single ram to generate CONF2 (n = 6) and OBF2 (n = 10). All mature F2 ewes were bred to a single ram to produce CONF3 (n = 6) and OBF3 (n = 10). OBF2 ewes exhibited greater (P < 0.0001) plasma cortisol than CONF2 throughout gestation. A glucose tolerance test at 90% gestation revealed OBF2 ewes had higher (P < 0.05) insulin response with similar glucose, resulting in greater (P < 0.05) insulin resistance. OBF3 neonates had similar weight, lean mass, and body fat mass to CONF3 neonates. These data suggest that multigenerational programming of adverse metabolic phenotypes occur in association with F0 maternal obesity, yet adiposity may return to CON levels in F3 neonates.

Three-dimensional microscale hanging drop arrays with geometric control for drug screening and live tissue imaging.

Existing three-dimensional (3D) culture techniques are limited by trade-offs between throughput, capacity for high-resolution imaging in living state, and geometric control. Here, we introduce a modular microscale hanging drop culture where simple design elements allow high replicates for drug screening, direct on-chip real-time or high-resolution confocal microscopy, and geometric control in 3D. Thousands of spheroids can be formed on our microchip in a single step and without any selective pressure from specific matrices. Microchip cultures from human LN229 glioblastoma and patient-derived mouse xenograft cells retained genomic alterations of originating tumors based on mate pair sequencing. We measured response to drugs over time with real-time microscopy on-chip. Last, by engineering droplets to form predetermined geometric shapes, we were able to manipulate the geometry of cultured cell masses. These outcomes can enable broad applications in advancing personalized medicine for cancer and drug discovery, tissue engineering, and stem cell research.

A broadly resolved molecular phylogeny of New Zealand cheilostome bryozoans as a framework for hypotheses of morphological evolution.

Larger molecular phylogenies based on ever more genes are becoming commonplace with the advent of cheaper and more streamlined sequencing and bioinformatics pipelines. However, many groups of inconspicuous but no less evolutionarily or ecologically important marine invertebrates are still neglected in the quest for understanding species- and higher-level phylogenetic relationships. Here, we alleviate this issue by presenting the molecular sequences of 165 cheilostome bryozoan species from New Zealand waters. New Zealand is our geographic region of choice as its cheilostome fauna is taxonomically, functionally and ecologically diverse, and better characterized than many other such faunas in the world. Using this most taxonomically broadly-sampled and statistically-supported cheilostome phylogeny comprising 214 species, when including previously published sequences, and 17 genes (2 nuclear and 15 mitochondrial) we tested several existing systematic hypotheses based solely on morphological observations. We find that lower taxonomic level hypotheses (species and genera) are robust while our inferred trees did not reflect current higher-level systematics (family and above), illustrating a general need for the rethinking of current hypotheses. To illustrate the utility of our new phylogeny, we reconstruct the evolutionary history of frontal shields (i.e., a calcified body-wall layer in ascus-bearing cheilostomes) and ask if its presence has any bearing on the diversification rates of cheilostomes.

Mindfulness-based stress reduction alters brain activity for breast cancer survivors with chronic neuropathic pain: preliminary evidence from resting-state fMRI.

PURPOSE: Breast cancer continues to be the most commonly diagnosed cancer among Canadian women, with as many as 25-60% of women suffering from chronic neuropathic pain (CNP) as a pervasive consequence of treatment. While pharmacological interventions have shown limited efficacy for the management of CNP to date, psychological interventions, such as mindfulness-based stress reduction (MBSR), may be a promising alterative for improving pain-related problems. The purpose of this study was to use brain imaging methods to investigate this potential. METHODS: Resting-state fMRI was used in female breast cancer survivors with CNP before and after an 8-week MBSR course (n = 13) and compared with a waitlist control group (n = 10). RESULTS: Focusing on the default mode network, the most significant results show greater posterior cingulate connectivity with medial prefrontal regions post-MBSR intervention. Moreover, this change in connectivity correlated with reduced pain severity for the MBSR group. CONCLUSIONS: These results provide empirical evidence of a change in the brain following MBSR intervention associated with changes in the subjective experience of pain. IMPLICATIONS FOR CANCER SURVIVORS: This study gives hope for a non-invasive method of easing the struggle of CNP in women following breast cancer treatment.

Million-year lag times in a post-orogenic sediment conveyor.

Understanding how sediment transport and storage will delay, attenuate, and even erase the erosional signal of tectonic and climatic forcings has bearing on our ability to read and interpret the geologic record effectively. Here, we estimate sediment transit times in Australia's largest river system, the Murray-Darling basin, by measuring downstream changes in cosmogenic 26Al/10Be/14C ratios in modern river sediment. Results show that the sediments have experienced multiple episodes of burial and reexposure, with cumulative lag times exceeding 1 Ma in the downstream reaches of the Murray and Darling rivers. Combined with low sediment supply rates and old sediment blanketing the landscape, we posit that sediment recycling in the Murray-Darling is an important and ongoing process that will substantially delay and alter signals of external environmental forcing transmitted from the sediment's hinterland.

Optimizing ophthalmic delivery of a poorly water soluble drug from an aqueous in situ gelling system.

Poorly soluble drugs are often unsuitable to incorporate in ocular in situ gelling systems due to the aqueous based gelling formulations and low volumes administered. For such formulations to be successful, the administered drug must have sufficient solubility to diffuse from the formulation to the eye and should not affect the gelation of the in situ gelling material. Drug salt forms can improve the solubility of poorly soluble drugs, however, as in situ gel forming formulations are often designed to be crosslinked by salts (present the lacrimal fluid) it can make salt forms difficult to formulate. The aim of this study was to develop an in situ gel forming ophthalmic formulation of a poorly soluble drug flurbiprofen (FBP) through cyclodextrin complex formation and to analyse the impact on gelation, release and permeation through the cornea. Hydroxypropyl-beta-cyclodextrin (HßCD) was used as a complexing agent and low acyl gellan gum was added to the FBP- HßCD complex as a water soluble in situ gelling polymer. Measurements were performed using rheo-dissolution, which utilises a rheometer with a modified lower plate that has the unique ability to allow rheological measurement and analysis of drug release simultaneously. An ex-vivo permeation study was also performed using porcine cornea. Rheological measurements in terms of elastic (G') and viscous (G″) modulus showed rapid gelation of the formulation upon contact with simulated lacrimal fluid (SLF). Approximately, 97% FBP was released when 10% HßCD was used and release was decreased to 79% when the amount of HßCD was increased to 20%. The percentage of drug permeation through the cornea was 55% in 300 min whereas the marketed non gelling eye drop formulation containing FBP sodium showed only 37% permeation. The data presented here, revealed that not only could a poorly soluble drug be complexed with cyclodextrin and loaded into an in situ gelling system without interfering with the gelation, but also permeability the of the drug improved.

Preindustrial 14CH4 indicates greater anthropogenic fossil CH4 emissions.

Atmospheric methane (CH4) is a potent greenhouse gas, and its mole fraction has more than doubled since the preindustrial era1. Fossil fuel extraction and use are among the largest anthropogenic sources of CH4 emissions, but the precise magnitude of these contributions is a subject of debate2,3. Carbon-14 in CH4 (14CH4) can be used to distinguish between fossil (14C-free) CH4 emissions and contemporaneous biogenic sources; however, poorly constrained direct 14CH4 emissions from nuclear reactors have complicated this approach since the middle of the 20th century4,5. Moreover, the partitioning of total fossil CH4 emissions (presently 172 to 195 teragrams CH4 per year)2,3 between anthropogenic and natural geological sources (such as seeps and mud volcanoes) is under debate; emission inventories suggest that the latter account for about 40 to 60 teragrams CH4 per year6,7. Geological emissions were less than 15.4 teragrams CH4 per year at the end of the Pleistocene, about 11,600 years ago8, but that period is an imperfect analogue for present-day emissions owing to the large terrestrial ice sheet cover, lower sea level and extensive permafrost. Here we use preindustrial-era ice core 14CH4 measurements to show that natural geological CH4 emissions to the atmosphere were about 1.6 teragrams CH4 per year, with a maximum of 5.4 teragrams CH4 per year (95 per cent confidence limit)-an order of magnitude lower than the currently used estimates. This result indicates that anthropogenic fossil CH4 emissions are underestimated by about 38 to 58 teragrams CH4 per year, or about 25 to 40 per cent of recent estimates. Our record highlights the human impact on the atmosphere and climate, provides a firm target for inventories of the global CH4 budget, and will help to inform strategies for targeted emission reductions9,10.

Old carbon reservoirs were not important in the deglacial methane budget.

Permafrost and methane hydrates are large, climate-sensitive old carbon reservoirs that have the potential to emit large quantities of methane, a potent greenhouse gas, as the Earth continues to warm. We present ice core isotopic measurements of methane (Δ14C, δ13C, and δD) from the last deglaciation, which is a partial analog for modern warming. Our results show that methane emissions from old carbon reservoirs in response to deglacial warming were small (<19 teragrams of methane per year, 95% confidence interval) and argue against similar methane emissions in response to future warming. Our results also indicate that methane emissions from biomass burning in the pre-Industrial Holocene were 22 to 56 teragrams of methane per year (95% confidence interval), which is comparable to today.

Mechanical and release behaviour of theophylline from matrix tablets containing psyllium powder in combination with grewia polysaccharides.

This study was aimed at investigating the effect of grewia polysaccharides on the mechanical and release properties of tablet matrices containing binary mixtures of the polysaccharide with psyllium. Two grades of grewia polysaccharides (GG and GDS) were extracted and binary mixtures of the polysaccharides with psyllium were formulated into tablet matrices containing theophylline as the model drug. The true, bulk and tapped densities, Carr's compressibility index of the powders and binary composites were determined before tablet compression. Tablet properties (hardness, porosity, and drug release from the matrices) were investigated. The dissolution test was carried out in 0.1 M HCl (pH 1.2) and phosphate buffer (pH 6.8). The results show that GG and GDS produced tablets with good mechanical strength (108.33 N and 95.70 N, respectively) while psyllium produced softer tablets (7.13 N). The combination of psyllium and grewia polysaccharides in the matrices resulted in a significant increase in the mechanical strength of the matrices when compared to matrices containing psyllium alone as the matrix former. The results also showed that GG and GDS reduced the dissolution rate and effectively eliminated the burst release of theophylline from the psyllium matrices at both pHs. The matrices of GG or GDS and the binary mixtures conform to non-Fickian anomalous diffusion with n > 0.45. When overcoming the burst release of drug from matrices such as psyllium, grewia polysaccharides may provide an effective reduction and a more sustained drug release from such matrices.

Rheo-dissolution: A new platform for the simultaneous measurement of rheology and drug release.

There is great potential to improve drug delivery through the use of in-situ gelling delivery systems. Here we demonstrate a technique capable of measuring changes in rheology (gelation and/or dissolution) of in-situ gelling delivery systems on contact with physiological fluid, while simultaneously analysing drug release. An ocular in-situ gelling formulation (gellan and timolol maleate) and an in-situ gelling oral liquid (alginate and metronidazole) were used as exemplar formulations. The method allowed profiling of increasing gellan concentration resulting in a reduction of timolol maleate released into simulated lacrimal fluid. When alginate was used as an in-situ gelling oral formulation there was a rapid increase in G' on contact with simulated gastric fluid. When this was changed to simulated intestinal fluid, drug release rate increased rapidly, coinciding with alginate gel dissolution. This work highlights the potential of this technology as a tool in development and optimisation of these increasingly popular delivery systems.

Radiocarbon Production Events and their Potential Relationship with the Schwabe Cycle.

Extreme cosmic radiation events occurred in the years 774/5 and 993/4 CE, as revealed by anomalies in the concentration of radiocarbon in known-age tree-rings. Most hypotheses point towards intense solar storms as the cause for these events, although little direct experimental support for this claim has thus far come to light. In this study, we perform very high-precision accelerator mass spectrometry (AMS) measurements on dendrochronological tree-rings spanning the years of the events of interest, as well as the Carrington Event of 1859 CE, which is recognized as an extreme solar storm even though it did not generate an anomalous radiocarbon signature. Our data, comprising 169 new and previously published measurements, appear to delineate the modulation of radiocarbon production due to the Schwabe (11-year) solar cycle. Moreover, they suggest that all three events occurred around the maximum of the solar cycle, adding experimental support for a common solar origin.

Elasticity and energy dissipation in the double network hydrogel adhesive of the slug Arion subfuscus.

The slug Arion subfuscus produces a mucus-based defensive secretion that is remarkably tough. This glue appears to be a double network hydrogel, gaining its toughness through the synergistic actions of two networks of polymers, a relatively stiff network and a relatively deformable network. The double network mechanism has great potential to guide the development of synthetic adhesives. Mechanical tests were performed to analyse key predictions of the mechanism. Stress relaxation tests and tensile tests support the presence of stable cross-links. Cyclic stress-strain tests demonstrate that the glue dissipates a great deal of energy through the failure of these cross-links as sacrificial bonds. Energy dissipation by failure of sacrificial bonds rather than viscous processes is supported by the minimal effect of the time course of the experiments on the measured properties. These sacrificial bonds appear able to reform within minutes after failure. Finally, the glue's stiffness decreases at pH values below 5.5, whereas magnesium and calcium rapidly dissociate from the glue at all pH values tested. Thus, these ions may not be the primary cross-linkers generating the glue's stiffness. This article is part of the theme issue 'Transdisciplinary approaches to the study of adhesion and adhesives in biological systems'.

The role of intraoperative cholangiography in patients undergoing laparoscopic cholecystectomy for acute gallstone pancreatitis: is magnetic resonance cholangiopancreatography needed?

INTRODUCTION: Intraoperative cholangiography is sporadically used in patients undergoing laparoscopic cholecystectomy to delineate common bile duct anatomy and exclude retained stones. In patients with acute gallstone pancreatitis, intraoperative cholangiography may reduce the need for preoperative magnetic resonance cholangiopancreatography. MATERIALS AND METHODS: A retrospective review of a prospectively collected patient database was undertaken over a 15-year period. The primary objective was to evaluate intraoperative assessment of the common bile duct with intraoperative cholangiography in patients with acute gallstone pancreatitis. RESULTS: A total of 2215 patients underwent laparoscopic cholecystectomy between October 1998 and December 2013; 113 patients (of whom 77 were women) with a mean age of 54 years (range 16-88 years) were diagnosed with acute gallstone pancreatitis. Of these, 102 patients (90%) underwent laparoscopic cholecystectomy with intraoperative cholangiography, which was normal in 89 cases. Thirteen patients had choledocholithiasis on intraoperative cholangiography, 11 of whom were managed with concomitant trans-cystic duct exploration and clearance. Two patients required postoperative endoscopic retrograde cholangiopancreatography. CONCLUSIONS: In patients diagnosed with acute gallstone pancreatitis, it is reasonable to proceed directly to surgery using intraoperative cholangiography on the same admission as the definitive assessment of the common bile duct. This negates the need for magnetic resonance cholangiopancreatography and can translate into cost savings and reduced length of stay.

Structure and physicochemical properties of Ghanaian grewia gum.

Grewia polysaccharides were isolated using sodium metabisulphite and phosphate buffers and the influence of the different extraction techniques on the chemical composition and structural characteristics of the extracts were determined. Structure and chemical composition of the resulting polysaccharide extracts were determined using FT-IR and NMR spectroscopy, neutral sugar analysis, size exclusion chromatography coupled to multi-angle light scattering (SEC-MALS), dilute solution viscometry and steady shear rheology. Chemical composition was similar irrespectively of the extraction solvent used and ranged between 11.1 and 16.5% for protein, 53.4 and 66.9% for total carbohydrate, 18.5 and 35.1% for total uronic acid and 23.5 and 28.6% for rhamnose. Predominate sugars in the extracts were rhamnose and uronic acids with spectroscopy showing the presence of esterified groups. Intrinsic viscosity varied between 6.5 and 9.1 dL g-1 and related with molar mass (754-2778 × 103 g mol-1). Grewia polysaccharide dispersions at 1 g dL-1 exhibited a shear thinning flow behaviour with crude and sodium metabisulphite extracts having higher viscosities. Overall, differences in extraction techniques produced grewia samples with tailored bulk properties for use in the food and pharmaceutical industries.

Mineral profiling of ostrich (Struthio camelus) seminal plasma and its relationship with semen traits and collection day.

Successful assisted reproduction techniques, with specific focus on in vitro semen storage for artificial insemination, are dependent on certain key elements which includes the biochemical profiling of semen. The objective of this study was to complete an ostrich seminal plasma (SP) evaluation by inductively coupled plasma mass spectrometry (ICP-MS) among seven males at different daily intervals (day 1, 3, 7, 11, 15, 19, 21, 23, 25, 26, 27, 28) for a period of 28 days during spring (August to September) for mineral profiling. The effect of collection day and male on sperm concentration, semen volume and seminal plasma volume, was explored as well as the relationships amongst these specific sperm traits and SP minerals. Variation amongst SP mineral concentrations, accounted for by the fixed effects of sperm concentration, semen volume, seminal plasma volume, collection day and male, ranged from 18% to 77%. Male had the largest effect on variation in SP minerals, namely: phosphorus (P), potassium (K), calcium (Ca), sodium (Na), boron (B), iron (Fe), cobalt (Co), nickel (Ni), copper (Cu), molybdenum (Mo), barium (Ba), arsenic (As) and selenium (Se). Sperm concentration instigated fluctuations of P, magnesium (Mg), B, zinc (Zn), Fe, aluminium (Al), Se, manganese (Mn) and lead (Pb). Semen volume had an effect on Na, K, B, Pb and Ba while seminal plasma volume only influenced variation in Na. There were fluctuations among collection days of specific micro minerals, Ni and Mo, with initial Ni concentrations being relatively greater and Mo at lesser concentrations. Semen volume, seminal plasma volume and sperm concentration varied amongst males. Sperm concentrations during the initial collection days, 1 and 3, were less than that for days 7 to 28. Significant variation of SP minerals and sperm characteristics among ejaculates and males suggest an association of these specific elements with sperm function and are, therefore, considered to be of potential importance to success of assisted reproduction technology for the ostrich. The relationship amongst sperm concentration and collection day confirms the need to conduct an initial period of collection to stabilise a greater sperm concentration to optimise sperm numbers for artificial insemination purposes.