Engineered aprotinin for improved stability of fibrin biomaterials.

Fibrin has been long used clinically for hemostasis and sealing, yet extension of use in other applications has been limited due to its relatively rapid resorption in vivo, even with addition of aprotinin or other protease inhibitors. We report an engineered aprotinin variant that can be immobilized within fibrin and thus provide extended longevity. When recombinantly fused to a transglutaminase substrate domain from α(2)-plasmin inhibitor (α(2)PI(1-8)), the resulting variant, aprotinin-α(2)PI(1-8), was covalently crosslinked into fibrin matrices during normal thrombin/factor XIIIa-mediated polymerization. Challenge with physiological plasmin concentrations revealed that aprotinin-α(2)PI(1-8)-containing matrices retained 78% of their mass after 3 wk, whereas matrices containing wild type (WT) aprotinin degraded completely within 1 wk. Plasmin challenge of commercial sealants Omrixil and Tisseel, supplemented with aprotinin-α(2)PI(1-8) or WT aprotinin, showed extended longevity as well. When seeded with human dermal fibroblasts, aprotinin-α(2)PI(1-8)-supplemented matrices supported cell growth for at least 33% longer than those containing WT aprotinin. Subcutaneously implanted matrices containing aprotinin-α(2)PI(1-8) were detectable in mice for more than twice as long as those containing WT aprotinin. We conclude that our engineered recombinant aprotinin variant can confer extended longevity to fibrin matrices more effectively than WT aprotinin in vitro and in vivo.

The effect of the year of wheat variety release on productivity and stability of performance on two organic and two non-organic farms

Nineteen wheat cultivars, released from 1934 to 2000, were grown at two organic and two non-organic sites in each of 3 years. Assessments included grain yield, grain protein concentration, protein yield, disease incidence and green leaf area. The superiority of each cultivar (the sum of the squares of the differences between its mean in each environment and the mean of the best cultivar there, divided by twice the number of environments; CS) was calculated for yield, grain protein concentration and protein yield, and ranked in each environment. The yield and grain protein concentration CS were more closely correlated with cultivar release date at the non-organic sites than at organic sites. This difference may be attributed to higher yield levels with larger differences among cultivars at the non-organic sites, rather than to improved stability (i.e. similar ranks) across sites. The significant difference in the correlation of protein yield CS and cultivar age between organic and non-organic sites would support evidence that the ability to take up mineral nitrogen (N) compared to soil N has been a component of the selection conditions of more modern cultivars (released after 1989). This is supported by assessment of green leaf area (GLA), where more modern cultivars in the non-organic systems had greater late-season GLA, a trend that was not identified in organic conditions. This effect could explain the poor correlation between age and protein yield CS in organic compared to non-organic conditions where modern cultivars are selected to benefit from later nitrogen (N) availability which includes the spring nitrogen applications tailored to coincide with peak crop demand. Under organic management, N release is largely based on the breakdown of fertility-building crops incorporated (ploughed-in) in the previous autumn. The release of nutrients from these residues is dependent on the soil conditions, which includes temperature and microbial populations, in addition to the potential leaching effect of high winter rainfall in the UK. In organic cereal crops, early resource capture is a major advantage for maximizing the utilization of nutrients from residue breakdown. It is concluded that selection of cultivars under conditions of high agrochemical inputs selects for cultivars that yield well under maximal conditions in terms of nutrient availability and pest, disease and weed control. The selection conditions for breeding have a tendency to select cultivars which perform relatively better in non-organic compared to organic systems.

Transient effects of stretching exercises on gait parameters of elderly women

This study aimed to analyse the effects of a single stretching exercise session on a number of gait parameters ill elderly participants in all attempt to determine whether these exercises can influence the risk of fall. Fifteen healthy women living in the community Volunteered to participate in the study. A kinematic gait analysis was performed immediately before and after a session of static stretching exercises applied oil hip flexor/extensor muscles. Results showed a significant influence of stretching exercises on a number of gait parameters, which have previously been proposed as fall predictors. Participants showed increased gait velocity, greater step length and reduced double Support time during stance after performing stretching exercises, suggesting improved stability and mobility. Changes around the pelvis (increased anterior-posterior tilt and rotation range of motion) resulting from the stretching exercises were suggested to influence the gait parameters (velocity, step length and double support time). Therefore, stretching exercises were shown to be a promising strategy to facilitate changes in gait parameters related to the risk of fall. Some other gait variables related to the risk of fall remained Unaltered (e.g., toe clearance). The stable pattern of segmental angular velocities was proposed to explain the stability of these unchanged gait variables. The results indicate that stretching exercises, performed oil a regular (daily) basis, result in gait adaptations which can be considered as indicative of reduced fall risk. Other Studies to determine whether regular stretching routines are an effective strategy to reduce the risk of fall are required. (C) 2008 Elsevier Ltd. All rights reserved.

Effects of Feed Time, Organic Loading and Shock Loads in Anaerobic Whey Treatment by an AnSBBR with Circulation

The aim of this work was to investigate the effect of different feeding times (2, 4, and 6 h) and organic loading rates (3, 6 and 12 gCOD l(-1) day(-1)) on the performance of an anaerobic sequencing batch reactor containing immobilized biomass, as well as to verify the minimum amount of alkalinity that can be added to the influent. The reactor, in which mixing was achieved by recirculation of the liquid phase, was maintained at 30 +/- 1A degrees C, possessed 2.5 l reactional volume and treated 1.5 l cheese whey in 8-h cycles. Results showed that the effect of feeding time on reactor performance was more pronounced at higher values of organic loading rates (OLR). During operation at an OLR of 3 gCOD l(-1) day(-1), change in feeding time did not affect efficiency of organic matter removal from the reactor. At an OLR of 6 gCOD l(-1) day(-1), reactor efficiency improved in relation to the lower loading rate and tended to drop at longer feeding times. At an OLR of 12 gCOD l(-1) day(-1) the reactor showed to depend more on feeding time; higher feeding times resulted in a decrease in reactor efficiency. Under all conditions shock loads of 24 gCOD l(-1) day(-1) caused an increase in acids concentration in the effluent. However, despite this increase, the reactor regained stability readily and alkalinity supplied to the influent showed to be sufficient to maintain pH close to neutral during operation. Regardless of applied OLR, operation with feeding time of 2 h was which provided improved stability and rendered the process less susceptible to shock loads.

Ultra-thin films of alternating semi-interpenetrating layers of a conducting polymer with thermosetting phenolic resins for sensor application

In this work a new method for crosslinking ultra-thin films with potential applications in sensor systems is proposed. The films were produced by layer-by-layer (LbL) assembly using a conducting polymer, poly(o-ethoxyaniline) (POEA), alternated with a thermosetting resin, novolac-type phenolformaldehyde (PF), crosslinked by a simple thermal treatment. The PF resin served as both alternating and crosslinking agents. The films were characterized by Fourier transform infrared (FTIR) and ultraviolet-visible (UV-Vis) spectroscopy, thermogravimetry (TG), desorption, doping/dedoping cycling and electrical measurements. The results showed that film architecture and crosslinking degree can be controlled by the conditions used for film deposition (number of bilayers, polymer concentration, pH, and deposition time), and crosslinking time. Moreover, this approach offers several advantages such as fast curing time and low cost, indicating that these films can be used to produce sensors with improved stability.

Subcycling first- and second-order generalizations of the trapezoidal rule

Subcycling algorithms which employ multiple timesteps have been previously proposed for explicit direct integration of first- and second-order systems of equations arising in finite element analysis, as well as for integration using explicit/implicit partitions of a model. The author has recently extended this work to implicit/implicit multi-timestep partitions of both first- and second-order systems. In this paper, improved algorithms for multi-timestep implicit integration are introduced, that overcome some weaknesses of those proposed previously. In particular, in the second-order case, improved stability is obtained. Some of the energy conservation properties of the Newmark family of algorithms are shown to be preserved in the new multi-timestep extensions of the Newmark method. In the first-order case, the generalized trapezoidal rule is extended to multiple timesteps, in a simple way that permits an implicit/implicit partition. Explicit special cases of the present algorithms exist. These are compared to algorithms proposed previously. (C) 1998 John Wiley & Sons, Ltd.

A numerical analysis of generalized Boussinesq-type equation using continuos/discontinuous FEM

An improved class of Boussinesq systems of an arbitrary order using a wave surface elevation and velocity potential formulation is derived. Dissipative effects and wave generation due to a time-dependent varying seabed are included. Thus, high-order source functions are considered. For the reduction of the system order and maintenance of some dispersive characteristics of the higher-order models, an extra O(mu 2n+2) term (n ??? N) is included in the velocity potential expansion. We introduce a nonlocal continuous/discontinuous Galerkin FEM with inner penalty terms to calculate the numerical solutions of the improved fourth-order models. The discretization of the spatial variables is made using continuous P2 Lagrange elements. A predictor-corrector scheme with an initialization given by an explicit RungeKutta method is also used for the time-variable integration. Moreover, a CFL-type condition is deduced for the linear problem with a constant bathymetry. To demonstrate the applicability of the model, we considered several test cases. Improved stability is achieved.

An Investigation of the Interstate 80 Rutting in Adair County, October 1983

This investigation was initiated to determine the causes of a rutting problem that occurred on Interstate 80 in Adair County. 1-80 from Iowa 25 to the Dallas County line was opened to traffic in November, 1960. The original pavement consisted of 4-1/2" of asphalt cement concrete over 12" of rolled stone base and 12" of granular subbase. A 5-1/2" overlay of asphalt cement concrete was placed in 1964. In 1970-1972, the roadway was resurfaced with 3" of asphalt cement concrete. In 1982, an asphalt cement concrete inlay, designed for a 10-year life, was placed in the eastbound lane. The mix designs for all courses met or exceeded all current criteria being used to formulate job mixes. Field construction reports indicate .that asphalt usage, densities, field voids and filler bitumen determinations were well within specification limits on a very consistent basis. Field laboratory reports indicate that laboratory voids for the base courses were within the prescribed limits for the base course and below the prescribed limits for the surface course. Instructional memorandums do indicate that extreme caution should be exercised when the voids are at or near the lower limits and traffic is not minimal. There is also a provision that provides for field voids controlling when there is a conflict between laboratory voids and field voids. It appears that contract documents do not adequately address the directions that must be taken when this conflict arises since it can readily be shown that laboratory voids must be in the very low or dangerous range if field voids are to be kept below the maximum limit under the current density specifications. A rut depth survey of January, 1983, identified little or no rutting on this section of roadway. Cross sections obtained in October, 1983, identified rutting which ranged from 0 to 0.9" with a general trend of the rutting to increase from a value of approximately 0.3" at MP 88 to a rut depth of 0.7" at MP 98. No areas of significant rutting were identified in the inside lane. Structural evaluation with the Road Rater indicated adequate structural capacity and also indicated that the longitudinal subdrains were functioning properly to provide adequate soil support values. Two pavement sections taken from the driving lane indicated very little distortion in the lower 7" base course. Essentially all of the distortion had occurred in the upper 2" base course and the 1..;1/2" surface course. Analysis of cores taken from this section of Interstate 80 indicated very little densification of either the surface or the upper or lower base courses. The asphalt cement content of both the Type B base courses and the Type A surface course were substantially higher than the intended asphalt cement content. The only explanation for this is that the salvaged material contained a greater percent of asphalt cement than initial extractions indicated. The penetration and viscosity of the blend of new asphalt cement and the asphalt cement recovered from the salvaged material were relatively close to that intended for this project. The 1983 ambient temperatures were extremely high from June 20 through September 10. The rutting is a result of a combination of adverse factors including, (1) high asphalt content, (2) the difference between laboratory and field voids, (3) lack of intermediate sized crushed particles, (4) high ambient temperatures. The high asphalt content in the 2" upper base course produced an asphalt concrete mix that did not exhibit satisfactory resistance to deformation from heavy loading. The majority of the rutting resulted from distortion of the 2" upper base lift. Heater planing is recommended as an interim corrective action. Further recommendation is to design for a 20-year alternative by removing 2-1/2" of material from the driving lane by milling and replacing with 2-1/2" of asphalt concrete with improved stability. This would be .followed by placing 1-1/2" of high quality resurfacing on the entire roadway. Other recommendations include improved density and stability requirements for asphalt concrete on high traffic roadways.

Asbestos Fibers in Type B Asphaltic Concrete Surface Course, MLR-67-1, 1968

The Standard Specifications for this project included requirements for placing two 500 foot test sections of Type B asphaltic concrete with 1-1/2 per cent asbestos fibres (mix size 3/8 inch, lift thickness 3/4 inch) as part of the regular construction of the surface course. These requirements were designed to provide asbestos modified mixtures for laboratory analysis and road performance evaluation. This report provides the preliminary results and analysis of test data obtained from tests on the mixtures placed on the roadway. Previous research by G. S. Zuelke (1) and J. H. Kestzman et al (2) indicated that asphaltic concrete mixtures modified with asbestos fibres improved stability, decreased permeability, and allowed the use of higher bitumen contents. This study indicated that the addition of asbestos fibres would permit the use of higher bitumen contents, theoretically improving durability, without adverse results. An indication was also obtained to the effect that asbestos mixtures were more difficult to compact in the field.

Assessment of Channelizing Device Effectiveness on High Speed/High Volume Roadways, 2007

Part 6 of the Manual on Uniform Traffic Control Devices (MUTCD) describes several types of channelizing devices that can be used to warn road users and guide them through work zones; these devices include cones, tubular markers, vertical panels, drums, barricades, and temporary raised islands. On higher speed/volume roadways, drums and/or vertical panels have been popular choices in many states, due to their formidable appearance and the enhanced visibility they provide when compared to standard cones. However, due to their larger size, drums also require more effort and storage space to transport, deploy and retrieve. Recent editions of the MUTCD have introduced new devices for channelizing; specifically of interest for this study is a taller (>36 inches) but thinner cone. While this new device does not offer a comparable target value to that of drums, the new devices are significantly larger than standard cones and they offer improved stability as well. In addition, these devices are more easily deployed and stored than drums and they cost less. Further, for applications previously using both drums and tall cones, the use of tall cones only provides the ability for delivery and setup by a single vehicle. An investigation of the effectiveness of the new channelizing devices provides a reference for states to use in selecting appropriate traffic control for high speed, high volume applications, especially for short term or limited duration exposures. This study includes a synthesis of common practices by state DOTs, as well as daytime and nighttime field observations of driver reactions using video detection equipment. The results of this study are promising for the day and night performance of the new tall cones, comparing favorably to the performance of drums when used for channelizing in tapers. The evaluation showed no statistical difference in merge distance and location, shy distance, or operating speed in either daytime or nighttime conditions. The study should provide a valuable resource for state DOTs to utilize in selecting the most effective channelizing device for use on high speed/high volume roadways where timely merging by drivers is critical to safety and mobility.

Evolution of Bioaffinity Reagents by Phage Display

Antibodies are natural binding proteins produced in vertebrates as a response to invading pathogens and foreign substances. Because of their capability for tight and specific binding, antibodies have found use as binding reagents in research and diagnostics. Properties of cloned recombinant antibodies can be further improved by means of in vitro evolution, combining mutagenesis with subsequent phage display selection. It is also possible to isolate entirely new antibodies from vast naïve or synthetic antibody libraries by phage display. In this study, library techniques and phage display selection were applied in order to optimise binding scaffolds and antigen recognition of antibodies, and to evolve new and improved bioaffinity reagents. Antibody libraries were generated by random and targeted mutagenesis. Expression and stability were mainly optimised by the random methods whereas targeted randomisation of the binding site residues was used for optimising the binding properties. Trinucleotide mutagenesis allowed design of defined randomisation patterns for a synthetic antibody library. Improved clones were selected by phage display. Capture by a specific anti- DHPS antibody was exploited in the selection of improved phage display of DHPS. Efficient selection for stability was established by combining phage display selection with denaturation under reducing conditions. Broad-specific binding of a generic anti-sulfonamide antibody was improved by selection with one of the weakest binding sulfonamides. In addition, p9 based phage display was studied in affinity selection from the synthetic library. A TIM barrel protein DHPS was engineered for efficient phage display by combining cysteinereplacement with random mutagenesis. The resulting clone allows use of phage display in further engineering of DHPS and possibly use as an alternative-binding scaffold. An anti-TSH scFv fragment, cloned from a monoclonal antibody, was engineered for improved stability to better suite an immunoassay. The improved scFv tolerates 8 – 9 °C higher temperature than the parental scFv and should have sufficient stability to be used in an immunoanalyser with incubation at 36 °C. The anti-TSH scFv fragment was compared with the corresponding Fab fragment and the parental monoclonal antibody as a capturing reagent in a rapid 5-min immunoassay for TSH. The scFv fragment provided some benefits over the conventionally used Mab in anayte-binding capacity and assay kinetics. However, the recombinant Fab fragment, which had similar kinetics to the scFv, provided a more sensitive and reliable assay than the scFv. Another cloned scFv fragment was engineered in order to improve broad-specific recognition of sulfonamides. The improved antibody detects different sulfonamides at concentrations below the maximum residue limit (100 μg/kg in EU and USA) and allows simultaneous screening of different sulfonamide drug residues. Finally, a synthetic antibody library was constructed and new antibodies were generated and affinity matured entirely in vitro. These results illuminate the possibilities of phage display and antibody engineering for generation and optimisation of binding reagents in vitro and indicate the potential of recombinant antibodies as affinity reagents in immunoassays.

Topical delivery of a naproxen-dithranol co-drug: in vitro skin penetration, permeation, and staining

Purpose This work probed the topical delivery and skin-staining properties of a novel co-drug, naproxyl-dithranol (Nap-DTH), which comprises anti-inflammatory (naproxen) and anti-proliferative (dithranol) moieties. Method Freshly excised, full-thickness porcine ear skin was dosed with saturated solutions of the compounds. After 24 h, the skin was recovered and used to prepare comparative depth profiles by the tape-stripping technique and to examine the extent of skin staining. Results Depth profiles showed that Nap-DTH led to a 5-fold increase in drug retention in the skin compared to dithranol. The application of Nap-DTH also demonstrated improved stability, resulting in lower levels of dithranol degradation products in the skin. Furthermore, significantly less naproxen from hydrolysed Nap-DTH permeated into the receptor phase compared to naproxen when applied alone (0.08 ± 0.03 nmol cm-² and 180 ± 60 nmol cm-², respectively). Moreover, the reduced staining of the skin was very apparent for Nap-DTH compared to dithranol. Conclusions Topical delivery of Nap-DTH not only improves the delivery of naproxen and dithranol, but also reduces unwanted effects of the parent moieties, in particular the skin staining, which is a major issue concerning the use of dithranol.

Carbon-decorated FePt nanoparticles

FePt magnetic nanoparticles are an important candidate material for many future magnetic applications. FePt exists as two main phases, that is, a disordered face-centered cubic (fcc) structure, which is generally prepared by chemical methods at low temperatures, and the high-temperature chemically ordered face-centered tetragonal (fct) structure. The fee FePt, with low coercivity but associated with superparamagnetic properties, may find applications as a magnetic fluid or as a nanoscale carrier for chemical or biochemical species in biomedical areas, while fct FePt is proposed for use in ultrahigh-density magnetic recording applications. However, for both of these applications an enhancement of the intrinsically weak magnetic properties, the avoidance of magnetic interferences from neighbor particles, and the improved stability of the small magnetic body remain key practical issues. We report a simple synthetic method for producing FePt nanoparticles that involves hydrothermal treatment of Fe and Pt precursors in glucose followed by calcination at 900 degrees C. This new method produces thermally stable spheroidal graphite nanoparticles (large and fullerene-like) that encapsulate or decorate FePt particles of ca. 5 nm with no severe macroscopic particle coalescence. Also, a low coercivity of the material is recorded; indicative of small magnetic interference from neighboring carbon-coated particles. Thus, this simple synthetic method involves the use of a more environmentally acceptable glucose/aqueous phase to offer a protective coating for FePt nanoparticles. It is also believed that such a synthetic protocol can be readily extended to the preparation of other graphite-coated magnetic iron alloys of controlled size, stoichiometry, and physical properties.

A new insight about pharmaceutical dosage forms for benzathine penicillin G

In this work, a micellar system of benzathine penicillin G (BPG) in sodium deoxycholate (NaDC) was developed and evaluated physicochemically. The solubility profile of the drug in water and buffer solutions at various pH was determined, as well as its n-octanol/water partition coefficient. The Critical Micellar Concentration of NaDC and its ability to incorporate BPG were also assessed. The study was carried out at low and high ionic strength which was adjusted by the addition of sodium chloride. The results demonstrated the ability of the micellar system to incorporate BPG, as well as to increase its apparent solubility in water. The enhancement of the solubility of BPG by the presence of NaDC micelles could be analyzed quantitatively within the framework of the pseudo-phase model. Concentration analysis showed that the micellar system could attain up to 90% incorporation of BPG. The incorporated drug is expected to exhibit improved stability, since the antibiotic enclosed in the hydrophobic core of micelles is rather shielded from the aqueous external environment

A new insight about pharmaceutical dosage forms for benzathine penicillin G

In this work, a micellar system of benzathine penicillin G (BPG) in sodium deoxycholate (NaDC) was developed and evaluated physicochemically. The solubility profile of the drug in water and buffer solutions at various pH was determined, as well as its n-octanol/water partition coefficient. The Critical Micellar Concentration of NaDC and its ability to incorporate BPG were also assessed. The study was carried out at low and high ionic strength which was adjusted by the addition of sodium chloride. The results demonstrated the ability of the micellar system to incorporate BPG, as well as to increase its apparent solubility in water. The enhancement of the solubility of BPG by the presence of NaDC micelles could be analyzed quantitatively within the framework of the pseudo-phase model. Concentration analysis showed that the micellar system could attain up to 90% incorporation of BPG. The incorporated drug is expected to exhibit improved stability, since the antibiotic enclosed in the hydrophobic core of micelles is rather shielded from the aqueous external environment.