In situ spatiotemporal characterization and analysis of chemical reactions using an ATR-integrated microfluidic reactor.

Determining kinetic reaction parameters with great detail has been of utmost importance in the field of chemical reaction engineering. However, commonly used experimental and computational methods however are unable to provide sufficiently resolved spatiotemporal information that can aid in the process of understanding these chemical reactions. With our work, we demonstrate the use of a custom designed single-bounce ATR-integrated microfluidic reactor to obtain spatiotemporal resolution for in situ monitoring of chemical reactions. Having a single-bounce ATR accessory allows us to individually address different sensing areas, thereby providing the ability to obtain spatially and temporally resolved information. To further enhance the spatial resolution, we utilize the benefits of synchrotron IR radiation with the smallest beam spot-size ∼150 µm. An on-flow modular microreactor additionally allows us to monitor the chemical reaction in situ, where the temporal characterization can be controlled with the operational flowrate. With a unique combination of experimental measurements and numerical simulations, we characterize and analyse a model SN2 reaction. For a chemical reaction between benzyl bromide (BB) and sodium azide (SA) to produce benzyl azide (BA), we successfully show the capability of our device to determine the diffusion coefficients of BB and SA as 0.367 ± 0.115 10-9 m2 s-1 and 1.17 ± 0.723 10-9 m2 s-1, respectively. Finally, with the above characteristics of our device, we also calculate a reaction rate of k = 0.0005 (m3s-1mol-1) for the given chemical reaction.

Power-free, digital and programmable dispensing of picoliter droplets using a Digit Chip.

There is a growing need for power-free methods to manipulate small volumes of liquids and thereby enable use of diagnostic assays in resource-limited settings. Most existing self-powered devices provide analog manipulation of fluids using paper, capillary or pressure-driven pumps. These strategies are well-suited to manipulating larger micro- and milliliter-scale volumes at constant flow rates; however, they fail to enable the manipulation of nanoliter and picoliter volumes required in assays using droplets, capillary sampling (e.g. finger prick), or expensive reagents. Here we report a device, termed the Digit Chip, that provides programmable and power-free digital manipulation of sub-nanoliter volumes. The device consists of a user-friendly button interface and a series of chambers connected by capillary valves that serve as digitization elements. Via a button press, the user dispenses and actuates ultra-small, quantitatively-programmed volumes. The device geometry is optimized using design models and experiments and precisely dispenses volumes as low as 21 pL with 97% accuracy. The volume dispensed can be tuned in 10 discrete steps across one order-of-magnitude with 98% accuracy. As a proof-of-principle that nanoliter-scale reagents can be precisely actuated and combined on-chip, we deploy the device to construct a precise concentration gradient with 10 discrete concentrations. Additionally, we apply this device alongside an inexpensive smartphone-based fluorescence imaging platform to perform a titration of E. coli with ampicillin. We observe the onset of bacterial death at a concentration of 5 µg mL-1, increasing to a maximum at 50 µg mL-1. These results establish the utility of the Digit Chip for diagnostic applications in low-resource environments.

How long do louse eggs take to hatch? A possible answer to an age-old riddle.

There are no rigorous data on how long eggs of the head louse, Pediculus capitis (Phthiraptera: Pediculidae), take to hatch. Pediculicide users often report reinfestations after apparently successful treatments in the absence of infective contacts. This study aimed to resolve the question of whether some louse eggs hatch after the completion of treatment, thereby giving rise to a new infestation. Data were extracted from the records of lice collected after treatments in 20 clinical intervention trials. All datasets were eliminated except those in which only newly hatched louse nymphs were found prior to the final assessment. This excluded the possibility that new eggs were laid after the first treatment and thus any young lice found must have originated from eggs laid before the start of treatment. This identified 23 of 1895 (1.2%) records with evidence of louse nymphs emerging at 13 days or more after the first treatment, 3­6 days longer than previous estimates. Current treatment regimens for pediculicides of two applications 7­10 days apart appear inadequate, which may explain continuing infestation in the community. Therefore, it is suggested that a revised approach using three treatments applied at intervals of 1 week should prevent the survival of any nymphs and their development into a new generation of adults.

Do nit removal formulations and other treatments loosen head louse eggs and nits from hair?

Eggs of the head louse, Pediculus capitis De Geer (Phthiraptera: Pediculidae), are difficult to remove because the female louse fixes them to hairs using a proteinaceous secretion that hardens within seconds. The persistent eggshells are harmless but unsightly and are often mistaken for an active infestation. Combing with a fine comb (nit comb) does not readily remove the eggs or empty eggshells because of the resilience of the fixative and both folk remedies and medical products have claimed to facilitate their removal. Measurement of the force required to initiate sliding of the egg fixative using a slip-peel tester was unable to detect evidence that any of three products which claimed to have egg-loosening properties (Step 2 Nit Removal System, Clear Lice Egg Remover, RID Lice Egg Loosener Gel) had any activity or exerted any effect on the egg fixative beyond the lubricating effects conveyed by water or conventional hair conditioner.

Compliant layer bearings in artificial joints. Part 2: simulator and fatigue testing to assess the durability of the interface between an elastomeric layer and a rigid substrate.

Artificial joints have been much improved since their introduction but they still have a limited lifetime. In an attempt to increase their life by improving the lubrication acting within these prostheses, compliant layered polyurethane (PU) joints have been devised. These joints mimic the natural synovial joint more closely by promoting fluid film lubrication. In this study, tests were performed on compliant layer joints to determine their ability to function under a range of conditions. Both static and dynamic compression tests were undertaken on compliant artificial hip joints of two different radial clearances. Friction tests were also performed before and after static loading. In addition to this, knee wear tests were conducted to determine the suitability of a compliant layer in these applications. In the knee tests, variations in experimental testing conditions were investigated using both active and passive rotation and severe malalignment of the tibial inserts. The static compression tests together with the friction studies suggest that a small radial clearance is likely to result in 'grabbing' contact between the head and cup. The larger radial clearance (0.33 microm) did not exhibit these problems. The importance of the design of the compliant layer joints was highlighted with delamination occurring on the lateral bearings during the knee wear studies. The bearings with a layer 2 mm thick performed better than the bearings with a layer 3 mm thick. Tests conducted on flat PU bearings resulted in no delamination; therefore, it was concluded that the layer separation was caused by design issues rather than by material issues. It was found that, with careful material choice, consideration of design, and effective manufacturing techniques, the compliant layer joint functioned well and demonstrated durability of the union between the hard and soft layers. These results give encouragement for the suitability of these joints for clinical use.

Compliant layer bearings in artificial joints. Part 1: the effects of different manufacturing techniques on the interface strength between an elastomeric layer and a rigid substrate.

The premise that elastomeric materials could be used as one or more of the articulating components in both hip and knee prostheses was postulated first by Unsworth and co-workers. It was thought that such materials might have the capacity to mimic natural joint behaviour more closely than the more rigid bearing surfaces commonly in use. A more natural joint function in artificial joints should promote better tribology, with full fluid-film lubrication being the goal. Early tests showed that this objective could potentially be achieved with a judicious choice of materials and carefully controlled manufacturing techniques. This paper (Part 1 of a two-part series) describes and explains the techniques used to verify the material selection as well as to determine the most appropriate manufacturing procedure to obtain a strong and robust interface between the support and bearing material of the prosthesis. Two polycarbonate urethane (PU) materials with different hardness values (Corethane 80A and Corethane 75D) gave sufficient interfacial strength when moulded under optimum conditions. Corethane 80A was used as the soft bearing material while Corethane 75D provided the rigid backing component. Peel tests revealed strong interface bonds, varying with processing conditions between 350 and 862 N. Fourier transform infrared spectroscopy and micro-thermal analysis showed that a fusion bond over 30 microm thick formed at the interface. The results of the range of tests and analyses, which have been used in this study, have provided sufficient evidence to validate the process used to manufacture these components.

Plastic detection comb better than visual screening for diagnosis of head louse infestation.

Finding lice can be difficult in head louse infestation. We compared a new louse detection comb with visual inspection. All children in two rural Turkish schools were screened by the two methods. Those with lice were offered treatment and the results monitored by detection combing. Children with nits only were re-screened to identify latent infestations. Using visual inspection we found 214/461 children (46%) with nits but only 30 (6.5%) with live lice. In contrast detection combing found 96 (21%) with live lice, of whom 20 had no nits. Detection combing was 3.84 times more effective than visual inspection for finding live lice. Only 10/138 (7.2%) children with nits and no lice were found to have active infestation by day 16. We found that the detection comb is significantly (P<0.001) more effective than visual screening for diagnosis; that nits are not a good indicator of active infestation; and that treatment with 1% permethrin was 89.6% effective.

Compliant-layer tibial bearing inserts: friction testing of different materials and designs for a new generation of prostheses that mimic the natural joint.

Total joint replacements (TJRs) have a limited lifetime, but the introduction of devices that exhibit good lubricating properties with low friction and low wear could well extend this. A novel tibial bearing design, using polyurethane (PU) as a compliant layer, to mimic the natural joint, has been developed. To determine accurately the mode of lubrication under which these joints operate, a synthetic lubricant was used in all these tests. Friction tests were carried out to assess the effects of material modulus and surface roughness, together with bearing design parameters such as bearing thickness and conformity, on lubrication. Corethane 80A was the preferred material and was chosen as the compliant layer for subsequent testing. A low surface roughness resulted in lower asperity contact as the asperities were depressed by the pressurized entraining fluid and full-fluid-film lubrication was approached. The three different tibial bearing conformities (low, medium, and high) did not appear to influence the mode of lubrication and all these bearings performed with extremely low friction. Similarly, the bearing thickness effects on lubrication at the levels tested (2 mm, 3 mm, and 4 mm) were minimal, although the effects of layer thickness on interface shear stress could be expected to be significant. This study describes a series of friction tests that have been used to select the most appropriate material and to optimize the design parameters to establish optimum conditions for these compliant layer joints.

DNA detection rates of host mtDNA in bloodmeals of human body lice (Pediculus humanus L., 1758).

Using polymerase chain reaction, we investigated the extent to which digestion affects the potential to amplify 12S mitochondrial DNA sequences from bloodmeals of individual human body lice (Pediculus humanus L.) (Phthiraptera, Pediculidae) up to 72 h after feeding on a surrogate rabbit host (Oryctolagus cuniculus L.) (Lagomorpha, Leporidae). Two rabbit-specific primer pairs were developed to produce amplicons of 199 bp and 283 bp, the smaller of which was found to have a significantly slower decay rate. Median detection periods (T50) for the amplicons were 20 h and 12 h, with maximum detection periods of 24 h and 12 h, respectively, suggesting an inversely proportional linear relationship between amplicon size and digestion time. The data provide an indication of timeframes essential for the design of forensic sampling protocols and a basis for investigating the feeding frequency of human lice.

Treatment of wine distillery wastewater by high rate anaerobic digestion.

Wine distillery wastewaters (WDW) are acidic and have a high content of potential organic pollutants. This causes high chemical oxygen demand (COD) values. Polyphenols constitute a significant portion of this COD, and limit the efficiency of biological treatment of WDWs. WDW starting parameters were as follows: pH 3.83, 4,185 mg/l soluble COD (COD(s)) and 674.6 mg/l of phenols. During operation, amendments of CaCO3 and K2HPO4, individually or in combination, were required for buffering the digester. Volatile fatty acid concentrations were < 300 mg/l throughout the study, indicating degradation of organic acids present. Mean COD(s) removal efficiency for the 130 day study was 87%, while the mean polyphenol, removal efficiency was 63%. Addition of 50 mg/l Fe(3+) between days 86 and 92 increased the removal efficiencies of COD(s) to 97% and of polyphenols to 65%. Addition of Co(3+) improved removal efficiencies to 97% for COD(s) and 92% for polyphenols. Optimization of anaerobic treatment was achieved at 30% WDW feed strength. Removal efficiencies of 92% and 84% were recorded at increased feed strength from days 108 to 130. High removal efficiencies of COD(s) and polyphenols after day 82 were attributed to the addition of macronutrients and micronutrients that caused pH stability and thus stimulated microbial activity.

An investigation and comparison of the bioactivity of selected essential oils on human lice and house dust mites.

The insecticidal potency of some essential oils suggests that they may find an application in the control of house dust mites, but current in vitro assays for mites do not appear to give consistent results. A simple, novel, mite chamber assay was therefore developed to carry out testing. Different species of insects are susceptible to different essential oil components, so we compared the relative acaricidal and pediculicidal activity of three essential oils: tea tree, lavender and lemon, because the activity of their constituents on lice ranges from highly active to virtually inactive. The most effective essential oil against both lice and mites was tea tree oil; lavender was the second most effective, and lemon oil the least, although it did show activity against mites, unlike lice. The assay proved simple and effective and gave reproducible results.

Compliant layer acetabular cups: friction testing of a range of materials and designs for a new generation of prosthesis that mimics the natural joint.

Total joint replacements (TJRs) have a limited lifetime, but the introduction of components that exhibit good lubricating properties with low friction and low wear could extend the life of TJRs. A novel acetabular cup design using polyurethane (PU) as a compliant layer (to mimic the natural joint) has been developed. This study describes a series of friction tests that have been used to select the most appropriate material, optimize the design parameters, and fine-tune the manufacturing processes of these joints. To determine accurately the mode of lubrication under which these joints operate, a synthetic lubricant was used in all these tests. Friction tests were carried out to assess the lubrication of four PU bearing materials. Corethane 80A was the preferred material and was subjected to subsequent testing. Friction tests conducted on acetabular cups, manufactured using Corethane 80A articulating against standard, commercially available femoral heads, demonstrated friction factors approaching those for full-fluid-film lubrication with only approximately 1 per cent asperity contact. As the joint produces these low friction factors within less than half a walking cycle after prolonged periods of loading, start-up friction was not considered to be a critical factor. Cups performed well across the full range of femoral head sizes, but a number of samples manufactured with reduced radial clearances performed with higher than expected friction. This was caused by the femoral head being gripped around the equator by the low clearance cup. To avoid this, the cup design was modified by increasing the flare at the rim. In addition to this the radial clearance was increased. As the material is incompressible, a radial clearance of 0.08 mm was too small for a cup diameter of 32 mm. A clearance of between 0.10 and 0.25 mm produced a performance approaching full-fluid-film lubrication. This series of tests acted as a step towards the optimization of the design of these joints, which has now led to an in vivo ovine model.

Lethality of essential oil constituents towards the human louse, Pediculus humanus, and its eggs.

Essential oils have been widely used in traditional medicine for the eradication of lice, including head lice, but due to the variability of their constitution the effects may not be reproducible. In an attempt to assess the contribution of their component monoterpenoids, a range of common individual compounds were tested in in vitro toxicity model against both human lice (Pediculus humanus, an accepted model of head lice lethality) and their eggs, at different concentrations. No detailed study into the relative potencies of their constituent terpenoids has so far been published. Adult lice were observed for lack of response to stimuli over 3 h and the LT(50) calculated, and the percentage of eggs failing to hatch was used to generate ovicidal activity data. A ranking was compiled for adult lice and partially for eggs, enabling structure-activity relationships to be assessed for lethality to both, and showed that, for activity in both life-cycle stages, different structural criteria were required. (+)-Terpinen-4-ol was the most effective compound against adult lice, followed by other mono-oxygenated monocyclic compounds, whereas nerolidol was particularly lethal to eggs, but ineffective against adult lice.

Design aspects of compliant, soft layer bearings for an experimental hip prosthesis.

Currently, an artificial hip joint can be expected to last, on average, in excess of 15 years with failure due, in the majority of cases, to late aseptic loosening of the acetabular component. A realistic alternative to the problem of wear in conventional joints is the introduction of bearing surfaces that exhibit low wear and operate in the full fluid-film lubrication regime. Contact analyses and friction tests were performed on compliant layer joints (metal-on-polyurethane) and the design of a prototype ovine arthroplasty model was investigated. When optimized, these components have been shown to achieve full fluid-film lubrication.

Influence of the electric field on a bio-mimetic film supported on a gold electrode.

A model biological membrane was formed by fusion of mixed cholesterol and DMPC (dimyristoylphosphatidylcholine) phospholipid vesicles onto a gold-coated quartz support. The gold surface was charged and the influence of the charge at the solid support on the structure and integrity of the phospholipid bilayer was investigated using the specular reflection of neutrons and electrochemical measurements. When the surface charge density is close to zero, the lipid vesicles fuse directly on the surface to form a bilayer with a small number of defects and hence low water content. When the support's surface is negatively charged the film swells and incorporates water due to the field driven poration of the membrane. When the charge density is more negative then -8 microC cm(-2) the bilayer is detached from the metal surface. However, it remains in close proximity to the metal electrode, suspended on a thin cushion of water. The film thicknesses, calculated from neutron reflectivity, have allowed us to determine the tilt angle of the lipid molecules as a function of the support's charge density. The lipid molecules are tilted 55 degrees from the surface normal at zero charge density but become significantly more perpendicular (30 degrees tilt angle) at charge densities more negative than -8 microC cm(-2). The tilt angle measurements are in very good agreement with previous IR studies. This paper describes the highlights of a more in-depth study which is fully described in [1].

Electric field-driven transformations of a supported model biological membrane--an electrochemical and neutron reflectivity study.

A mixed bilayer of cholesterol and dimyristoylphosphatidylcholine has been formed on a gold-coated block of quartz by fusion of small unilamellar vesicles. The formation of this bilayer lipid membrane on a conductive surface allowed us to study the influence of the support's surface charge on the structure and hydration of the bilayer lipid membrane. We have employed electrochemical measurements and the specular reflection of neutrons to measure the thickness and water content in the bilayer lipid membrane as a function of the charge on the support's surface. When the surface charge density is close to zero, the lipid vesicles fuse directly on the surface to form a bilayer with a small number of defects and hence small water content. When the support's surface is negatively charged the film swells and incorporates water. When the charge density is more negative than -8 micro C cm(-2), the bilayer starts to detach from the metal surface. However, it remains in a close proximity to the metal electrode, being suspended on a thin cushion of the electrolyte. The field-driven transformations of the bilayer lead to significant changes in the film thicknesses. At charge densities more negative than -20 micro C cm(-2), the bilayer is approximately 37 A thick and this number is comparable to the thickness determined for hydrated multilayers of dimyristoylphosphatidylcholine from x-ray diffraction experiments. The thickness of the bilayer decreases at smaller charge densities to become equal to approximately 26 A at zero charge. This result indicates that the tilt of the acyl chains with respect to the bilayer normal changes from approximately 35 degrees to 59 degrees by moving from high negative charges (and potentials) to zero charge on the metal.

Long-term results for Kinemax and Kinematic knee bearings on a six-station knee wear simulator.

A long-term wear test was performed on Kinemax and Kinematic (Howmedica Inc.) knee bearings on the Durham six-station knee wear simulator. The bearings were subjected to flexion/extension of 65-0 degrees, anterior-posterior translation of between 4.5 and 8.5 mm and a maximum axial load of 3 kN. Passive abduction/adduction and internal/external rotation were also permitted, however, two of the stations had a linkage system which produced +/- 5 degrees active internal/external rotation. The bearings were tested at 37 degrees C in a 30 per cent bovine serum solution and the test was run to 5.6 x 10(6) cycles. The bearings from stations 2 and 3, and stations 4 and 5 were swapped during the test to investigate the effects of interstation variability. The average wear rate and standard error was 3.00 +/- 0.98 mg/10(6) cycles (range 1.33-4.72 mg/10(6) cycles) for the Kinemax bearings and 3.78 +/- 1.04 mg/10(6) cycles (range 1.87-4.89 mg/10(6) cycles) for the Kinematic bearings. There were no significant differences in wear rates between the different bearing designs, the addition of active internal/external rotation or a change of stations. However, the wear tracks were different for the two types of bearings and with active internal/external rotation. The wear rates and factors were generally lower than previously published in vitro wear results; however, this may have been due to a difference in the axial loads and lubricants used. The appearance of the wear tracks with active internal/external rotation was comparable with those seen on explanted knee bearings.

Evaluation of a hip joint simulator.

To evaluate the functioning of the Durham hip joint wear simulator, the wear rates of ultra high molecular weight polyethylene (UHMWPE) and polytetrafluoroethylene (PTFE) acetabular cups articulating against 22 mm diameter cobalt-chromium-molybdenum (CoCrMo) femoral heads were studied. A wear test was conducted in a lubricant of distilled water at 37 degrees C for a duration of 4.8 million cycles. The average penetration rate for the CoCrMo femoral heads against UHMWPE acetabular cups was 0.03 mm/10(6) cycles, while penetration rate for PTFE cups was some twenty times greater. These results are of a similar order of magnitude to other simulator studies in distilled water and are in a similar ratio to clinical data.