A non-destructive test to assess the axial heterogeneity of in situ modified monoliths for HPLC

This paper describes a non-destructive "peak parking" protocol in order to assess the axial heterogeneity of an in situ modified monolithic column for high performance liquid chromatography; a "gradient stationary phase" was designed whereby the ligand density decreases along the length of the rod in the "forward flow" configuration. The results of multi-location peak parking demonstrated a consistent increase in peak variance from the 1 cm position of the column to the 9 cm location. This increase in band broadening supported the theory of a decreasing ligand density along the length of this gradient column. This is consistent with efficiency measurements performed in both the forward and reverse flow directions, with an improved efficiency (15% increase in N m^-1) in the reverse direction. These results are consistent with theoretical investigations into stationary phase gradients.

In-situ investigation of twinning behaviour in Mg-3Al-1Zn

Magnesium alloys are attractive for automotive and aerospace industries, due to their low density. One problem with these alloys is their limited formability at room temperature. Twinning plays a dominant role in deformation behaviour and it can be expected that an increased understanding of twinning will help improve formability. In the present work, the behaviour of different twinning systems in as-cast Mg-3AI-IZn is investigated using in-situ tensile tests in a scanning electron microscope. Electron backscatter diffraction and back scatter electron imaging were carried out during the tests. The results show both "tension" and "compression" twinning are active at room temperature and that twinning and untwinning occur both during loading and unloading.

In situ growth of Soletellina alba (Bivalvia: Psammobiidae) in response to detrital supply and mouth status in a seasonally-closed estuary

The supply of detritus is an important food source for many soft-sediment invertebrates, but its importance for their growth and condition is rarely, if ever, tested directly using manipulative field experiments. Therefore, we designed such a study to: (1) test the importance of fine particulate organic matter for the growth and condition of the infaunal bivalve Soletellina alba; (2) indirectly test the feeding mode of S. alba, which has been assumed to be a deposit feeder like other members of the same superfamily (Tellinoidea); (3) compare growth rates across two summers with contrasting patterns of estuary mouth opening/closing; and (4) compare the condition of individuals used in two field studies (i.e. present versus past) and a past laboratory study. Neither growth nor condition differed when organic content of the sediments was varied, which suggests that S. alba is either a suspension feeder or capable of switching modes of feeding. There was considerable interannual variation in growth with greater growth occurring during the summer with a longer period of mouth opening. This suggests that periods of mouth closure may reduce secondary production within seasonally-closed estuaries. Potential artefacts associated with laboratory trials were also identified, with laboratory bivalves exhibiting poorer condition than those used in two field trials. The present study provides no evidence that variable quantities and qualities of organic matter within the sediments influence the growth and condition of S. alba, but future studies should focus on food supplied via the water column when the estuary is open versus closed.

Effect of thermomechanical processing on the microstructure and retained austenite stability during in situ tensile testing using synchrotron X-Ray diffraction of NbMoAl TRIP steel

In this work we compare and contrast the stability of retained austenite during tensile testing of Nb-Mo-Al transformation-induced plasticity steel subjected to different thermomechanical processing schedules. The obtained microstructures were characterised using optical metallography, transmission electron microscopy and X-ray diffraction. The transformation of retained austenite to martensite under tensile loading was observed by in-situ high energy X-ray diffraction at 1ID / APS. It has been shown that the variations in the microstructure of the steel, such as volume fractions of present phases, their morphology and dimensions, play a critical role in the strain-induced transition of retained austenite to martensite.

An in situ method for the study of strain broadening using synchrotron X-ray diffraction

A tensonometer for stretching metal foils has been constructed for the study of strain broadening in X-ray diffraction line profiles. This device, which is designed for use on powder diffractometers and was tested on Station 2.3 at Daresbury Laboratory, allows in situ measurements to be performed on samples under stress. It can be used for data collection in either transmission or reflection modes using either symmetric or asymmetric diffraction geometries. As a test case, measurements were carried out on an 18 µm-thick copper foil experiencing strain levels of up to 5% using both symmetric reflection and symmetric transmission diffraction. All the diffraction profiles displayed peak broadening and asymmetry which increased with strain. The measured profiles were analysed by the fundamental-parameters approach using the TOPAS peak-fitting software. All the observed broadened profiles were modelled by convoluting a refineable diffraction profile, representing the dislocation and crystallite size broadening, with a fixed instrumental profile predetermined using high-quality LaB6 reference powder. The deconvolution process yielded `pure' sample integral breadths and asymmetry results which displayed a strong dependence on applied strain and increased almost linearly with applied strain. Assuming crystallite size broadening in combination with dislocation broadening arising from f.c.c. a/2〈110〉{111} dislocations, the variation of mechanical property with strain has been extracted. The observation of both peak asymmetry and broadening has been interpreted as a manifestation of a cellular structure with cell walls and cell interiors possessing high and low dislocation densities.

A systematic study of carbon fibre surface grafting via in situ diazonium generation for improved interfacial shear strength in epoxy matrix composites

A recently established means of surface functionalization of unsized carbon fibres for enhanced compatibility with epoxy resins was optimised and evaluated using interfacial shear stress measurements. Interfacial adhesion has a strong influence on the bulk mechanical properties of composite materials. In this work we report on the optimisation of our aryl diazo-grafting methodology via a series of reagent concentration studies. The fibres functionalised at each concentration are characterised physically (tensile strength, modulus, coefficient of friction, and via AFM), and chemically (XPS). The interfacial shear strength (IFSS) of all treated fibres was determined via the single fibre fragmentation test, using the Kelly-Tyson model. Large increases in IFSS for all concentrations (28-47%) relative to control fibres were observed. We show that halving the reagent concentration increased the coefficient of friction of the fibre and the interfacial shear strength of the composite while resulting in no loss of the key performance characteristics in the treated fibre.

In situ measurement of pipeline coating integrity and corrosion resistance losses under simulated mechanical strains and cathodic protection

A novel experimental assembly consisting of a specially designed tensile testing rig and a standard electrochemical flat cell has been designed for simulating buried high pressure pipeline environmental conditions in which a coating gets damaged and degrades under mechanical strain, and for studying the influence of mechanically induced damages such as the cracking of a coating on its anti-corrosion property. The experimental assembly is also capable of applying a cathodic protection (CP) potential simultaneously with the mechanical strain and environmental exposure. The influence of applied mechanical strain as well as extended exposure to the corrosive environment, coupled with the application of CP, has been investigated based on changes in electrochemical impedance spectroscopy (EIS). Preliminary results show that the amplitude of the coating impedance decreases with an increase in the applied strain level and the length of environmental exposure. The EIS characteristics and changes are found to correlate well with variations in coating cracking and degradation features observed on post-test samples using both optical microscopy and scanning electron microscopy. These results demonstrate that this new experimental method can be used to simulate and examine coating behaviour under the effects of complex high pressure pipeline mechanical, electrochemical and environmental conditions.

In situ localization associates biologically active plant natriuretic peptide immuno-analogues with conductive tissue and stomata

Plant natriuretic peptide immuno-analogues (irPNP) have previously been shown to affect a number of biological processes including stomatal guard cell movements, ion fluxes and osmoticum-dependent water transport. Tissue printing and immunofluorescent labelling techniques have been used here to study the tissue and cellular localization of irPNP in ivy (Hedera helix L.) and potato (Solanum tuberosum L.). Polyclonal antibodies active against human atrial natriuretic peptide (anti-hANP) and antibodies against irPNP from potato (anti-StPNP) were used for immunolabelling. Tissue prints revealed that immunoreactants are concentrated in vascular tissues of leaves, petioles and stems. Phloem-associated cells, xylem cells and parenchymatic xylem cells showed the strongest immunoreaction. Immunofluorescent microscopy with fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG supported this finding and, furthermore, revealed strong labelling to stomatal guard cells and the adjacent apoplastic space as well. Biologically active immunoreactants were also detected in xylem exudates of a soft South African perennial forest sage (Plectranthus ciliatus E. Mey ex Benth.) thus strengthening the evidence for a systemic role of the protein. In summary, in situ cellular localization is consistent with physiological responses elicited by irPNPs reported previously and is indicative of a systemic role in plant homeostasis.

Lupin kernel fiber consumption modifies fecal microbiota in healthy men as determined by rRNA gene flourescent in situ by hybridization

Background Changes in the composition of gastrointestinal microbiota by dietary interventions using pro- and prebiotics provide opportunity for improving health and preventing disease. However, the capacity of lupin kernel fiber (LKFibre), a novel legume-derived food ingredient, to act as a prebiotic and modulate the colonic microbiota in humans needed investigation.

Aim of the study The present study aimed to determine the effect of LKFibre on human intestinal microbiota by quantitative fluorescent in situ hybridization (FISH) analysis.

Design A total of 18 free-living healthy males between the ages of 24 and 64 years consumed a control diet and a LKFibre diet (containing an additional 17–30 g/day fiber beyond that of the control—incorporated into daily food items) for 28 days with a 28-day washout period in a single-blind, randomized, crossover dietary intervention design.
Methods Fecal samples were collected for 3 days towards the end of each diet and microbial populations analyzed by FISH analysis using 16S rRNA gene-based oligonucleotide probes targeting total and predominant microbial populations.

Results Significantly higher levels of Bifidobacterium spp. (P = 0.001) and significantly lower levels of the clostridia group of C. ramosum, C. spiroforme and C. cocleatum (P = 0.039) were observed on the LKFibre diet compared with the control. No significant differences between the LKFibre and the control diet were observed for total bacteria, Lactobacillus spp., the Eubacterium spp., the C. histolyticum/C. lituseburense group and the Bacteroides–Prevotella group.
Conclusions Ingestion of LKFibre stimulated colonic bifidobacteria growth, which suggests that this dietary fiber may be considered as a prebiotic and may beneficially contribute to colon health.

Sexual conflict and polyspermy under sperm-limited conditions : in situ evidence from field simulations with the free-spawning marine echinoid Evechinus chloroticus

For free‐spawning organisms that release gametes into the sea, sperm limitation (too few sperm to fertilize all eggs) is a major factor limiting reproductive success. Given such circumstances, the presence of several mechanisms to prevent polyspermy (too many sperm) may seem paradoxical; however, a growing body of data suggests that natural fertilization levels, though variable, can routinely be high. Under such conditions, polyspermy is much more likely. The tension between sperm limitation and polyspermy represents sexual conflict because males, in competing to fertilize as many eggs as possible, can impose lethal costs on eggs if multiple sperm gain entry. Here we present data for a marine invertebrate indicating high levels of polyspermy under sperm‐limited conditions. When the sea urchin Evechinus chloroticus was induced to spawn in situ, mean rates of polyspermy were 17.3% ±3.4%, and polyspermy was recorded at rates as high as 62.7%. Polyspermy was nearly always present, even when fertilization rates were <50%, confirming predictions that it should be present under sperm‐limited conditions. Both sperm limitation and polyspermy imposed substantial reproductive costs, and we conclude that both sexual conflict related to polyspermy and sperm limitation have been simultaneous strong selective forces shaping the evolution of reproductive traits in the sea.

In situ formation of BN nanotubes during nitriding reactions

High-yield multiwalled boron nitride (BN) nanotubes have been produced using a ball milling-annealing method. The BN nanotubes with a diameter less than 10 nm and a well-crystallized multiwalled structure were formed via an in situ nitriding reaction. The systematic investigation of the formation process at different annealing temperatures and for different times suggested that the formation of the unique multiwalled structure was attributed by a two-dimensional growth of the BN phase and a nonmetal catalytic growth.

Investigation of deformation mechanisms involved in the plasticity of AZ31 Mg alloy: in situ neutron diffraction and EPSC modelling

In situ neutron diffraction and Elasto-Plastic Self-Consistent (EPSC) polycrystal modelling have been employed to investigate which deformation mechanisms are involved in the plasticity of extruded AZ31 Mg alloy during the tensile loading along the extrusion direction. On the basis of this study we were able to determine the relative activity of the slip and twinning deformation modes. By tuning the parameters of the EPSC model (i.e. the critical resolved shear strengths and hardening parameters), excellent agreement with the experimental data has been achieved. It is shown that the strain in the crystallographic ⟨c ⟩direction is accommodated mainly by ⟨c + a ⟩ dislocation slip on second-order pyramidal planes. The results further indicate that either slip of ⟨a ⟩dislocations occurs on {10.1} pyramidal planes or cross-slip from basal and prismatic planes takes place.

Local nitric oxide synthase inhibition reduces skeletal muscle glucose uptake but not capillary blood flow during in situ muscle contraction in rats

OBJECTIVE: We have previously shown in humans that local infusion of a nitric oxide synthase (NOS) inhibitor into the femoral artery attenuates the increase in leg glucose uptake during exercise without influencing total leg blood flow. However, rodent studies examining the effect of NOS inhibition on contraction-stimulated skeletal muscle glucose uptake have yielded contradictory results. This study examined the effect of local infusion of an NOS inhibitor on skeletal muscle glucose uptake (2-deoxyglucose) and capillary blood flow (contrast-enhanced ultrasound) during in situ contractions in rats.

RESEARCH DESIGN AND METHODS: Male hooded Wistar rats were anesthetized and one hindleg electrically stimulated to contract (2 Hz, 0.1 ms) for 30 min while the other leg rested. After 10 min, the NOS inhibitor NG-nitro-L-arginine methyl ester (L-NAME) (arterial concentration of 5 µmol/l) or saline was infused into the epigastric artery of the contracting leg.

RESULTS: Local NOS inhibition had no effect on blood pressure, heart rate, or muscle contraction force. Contractions increased (P < 0.05) skeletal muscle NOS activity, and this was prevented by L-NAME infusion. NOS inhibition caused a modest significant (P < 0.05) attenuation of the increase in femoral blood flow during contractions, but importantly there was no effect on capillary recruitment. NOS inhibition attenuated (P < 0.05) the increase in contraction-stimulated skeletal muscle glucose uptake by ~35%, without affecting AMP-activated protein kinase (AMPK) activation.

CONCLUSIONS: NOS inhibition attenuated increases in skeletal muscle glucose uptake during contraction without influencing capillary recruitment, suggesting that NO is critical for part of the normal increase in skeletal muscle fiber glucose uptake during contraction.