Resistance of porcine blood clots to lysis relates to poor activation of porcine plasminogen by tissue plasminogen activator

In-vitro experimentation was performed on porcine and human blood to determine their comparative responsiveness to a novel fibrinolytic inhibitor and thereby assess whether the pig is a suitable animal model for subsequent in-vivo testing of this inhibitor. Thromboelastography showed the clots formed from porcine whole blood to be highly resistant to tissue plasminogen activator (t-PA)-catalyzed lysis, and this communication offers the resistance of porcine plasminogen to activation by t-PA as an explanation. Porcine blood containing 100 and 1500 IU/ml added t-PA lysed very slowly, having LY30 values of 1.9 +/- 1.4 and 2.9 +/- 1.9%, respectively. In contrast, the LY30 values for the human clots containing 100 and 1500 IU/ml t-PA were 77.1 +/- 6.3 and 93.3 +/- 1.3%, respectively. Moreover, purified porcine plasminogen was activated very slowly by added t-PA in the presence of both human and porcine fibrin. Activation of plasminogen by the endogenous activators, as measured by the euglobulin clot lysis time, was greatly prolonged for the pig (22 +/- 3 h) compared with the human (3.5 +/- 1.5 h). These results suggest caution in using the pig as an experimental model when studying the effects of various agents on fibrinolysis.

Modelling the gastric epithelium for testing of new chemical entities

A cell culture model of the gastric epithelial cell surface would prove useful for biopharmaceutical screening of new chemical entities and dosage forms. A successful model should exhibit tight junction formation, maintenance of differentiation and polarity. Conditions for primary culture of guinea-pig gastric mucous epithelial cell monolayers on Tissue Culture Plastic (TCP) and membrane insects (Transwells) were established. Tight junction formation for cells grown on Transwells for three days was assessed by measurement of transepithelial resistance (TEER) and permeability of mannitol and fluorescein. Coating the polycarbonate filter with collagen IV, rather with collagen I, enhanced tight junction formation. TEER for cells grown on Transwells coated with collagen IV was close to that obtained with intact guinea-pig gastric epithelium in vitro. Differentiation was assessed by incorporation of [3H] glucosamine into glycoprotein and by activity of NADPH oxidase, which produces superoxide. Both of these measures were greater for cells grown on filters coated with collagen I than for cells grown on TCP, but no major difference was found between cells grown on collagens I and IV. However, monolayers grown on membranes coated with collagen IV exhibited apically polarized secretion of mucin and superoxide. The proportion of cells, which stained positively for mucin with periodic Schiff reagent, was greater than 95% for all culture conditions. Gastric epithelial monolayers grown on Transwells coated with collagen IV were able to withstand transient (30 min) apical acidification to pH 3, which was associated with a decrease in [3H] mannitol flux and an increase in TEER relative to pH 7.4. The model was used to provide the first direct demonstration that an NSAID (indomethacin) accumulated in gastric epithelial cells exposed to low apical pH. In conclusion, guinea-pig epithelial cells cultured on collagen IV represent a promising model of the gastric surface epithelium suitable for screening procedures.

Abrasion resistance of fibre reinforced concrete floors

This thesis focuses on the investigation of the abrasion resistance of fibre reinforced concrete floors at both the macro and micro levels. A literature review of the available literature concerning subjects allied to the current project is included. This highlights themes relevant to wear mechanisms and the factors influencing it: factors that affect the abrasion resistance of concrete and several test methods for assessing it; and the historical development of fibres and the properties of different fibre types and their influence on concrete. Three accelerated abrasion testers were compared and critically discussed for their suitability for assessing the abrasion resistance of concrete floors. Based on the experimental findings one accelerated abrasion apparatus was selected as more appropriate to be used for carrying out the main investigations. The laboratory programme that followed was undertaken to investigate the influence of various material and construction factors on abrasion resistance. These included mix variations (w/c ratio), fibre reinforcement, geometry, type and volume, curing method and superplasticizing agents. The results clearly show that these factors significantly affected abrasion resistance and several mechanisms were presumed to explain and better understand these observations. To verify and understand these mechanisms that are accountable for the breakdown of concrete slabs, the same concrete specimens that were used for the macro-study, were also subjected to microstructutural investigations using techniques such as Microhardness examination, Mercury intrusion porosimetry and Petrographic examination. It has been found that the abrasion resistance of concrete is primarily dependent on the microstructure and porosity of the concrete nearest to the surface. The feasibility of predicting the abrasion resistance of fibre reinforced concrete floors by indirect and non-destructive methods was investigated using five methods that have frequently been used for assessing the quality of concrete. They included the initial surface absorption test, the impact test, ball cratering, the scratch test and the base hardness test. The impact resistance (BRE screed tester) and scratch resistance (Base hardness tester) were found to be the most sensitive to factors affecting abrasion resistance and hence are considered to be the most appropriate testing techniques. In an attempt to develop an appropriate method for assessing the abrasion resistance of heavy-duty industrial concrete floors, it was found that the presence of curing/sealing compound on the concrete surface at the time of accelerated abrasion testing produces inappropriate results. A preliminary investigation in the direction of modifying the Aston accelerated abrasion tester has been carried out and a more aggressive head has been developed and is pending future research towards standardisation.

The Effect of low-grade aggregates on the abrasion resistance of concrete

Three types of crushed rock aggregate were appraised, these being Carboniferous Sandstone, Magnesian Limestone and Jurassic Limestone. A comprehensive aggregate testing programme assessed the properties of these materials. Two series of specimen slabs were cast and power finished using recognised site procedures to assess firstly the influence of these aggregates as the coarse fraction, and secondly as the fine fraction. Each specimen slab was tested at 28 days under three regimes to simulate 2-body abrasion, 3-body abrasion and the effect of water on the abrasion of concrete. The abrasion resistance was measured using a recognised accelerated abrasion testing apparatus employing rotating steel wheels. Relationships between the aggregate and concrete properties and the abrasion resistance have been developed with the following properties being particularly important - Los Angeles Abrasion and grading of the coarse aggregate, hardness of the fine aggregate and water-cement ratio of the concrete. The sole use of cube strength as a measure of abrasion resistance has been shown to be unreliable by this work. A graphical method for predicting the potential abrasion resistance of concrete using various aggregate and concrete properties has been proposed. The effect of varying the proportion of low-grade aggregate in the mix has also been investigated. Possible mechanisms involved during abrasion have been discussed, including localised crushing and failure of the aggregate/paste bond. Aggregates from each of the groups were found to satisfy current specifications for direct finished concrete floors. This work strengthens the case for the increased use of low-grade aggregates in the future.

Negative input-resistance compensator for a constant power load

A negative input-resistance compensator is designed to stabilize a power electronic brushless dc motor drive with constant power-load characteristics. The strategy is to feed a portion of the changes in the dc-link voltage into the current control loop to modify the system input impedance in the midfrequency range and thereby to damp the input filter. The design process of the compensator and the selection of parameters are described. The impact of the compensator is examined on the motor-controller performance, and finally, the effectiveness of the controller is verified by simulation and experimental testing.

Integrated surface-ground water modeling in wetlands with improved methods to simulate vegetative resistance to flow

In topographically flat wetlands, where shallow water table and conductive soil may develop as a result of wet and dry seasons, the connection between surface water and groundwater is not only present, but perhaps the key factor dominating the magnitude and direction of water flux. Due to their complex characteristics, modeling waterflow through wetlands using more realistic process formulations (integrated surface-ground water and vegetative resistance) is an actual necessity. This dissertation focused on developing an integrated surface – subsurface hydrologic simulation numerical model by programming and testing the coupling of the USGS MODFLOW-2005 Groundwater Flow Process (GWF) package (USGS, 2005) with the 2D surface water routing model: FLO-2D (O’Brien et al., 1993). The coupling included the necessary procedures to numerically integrate and verify both models as a single computational software system that will heretofore be referred to as WHIMFLO-2D (Wetlands Hydrology Integrated Model). An improved physical formulation of flow resistance through vegetation in shallow waters based on the concept of drag force was also implemented for the simulations of floodplains, while the use of the classical methods (e.g., Manning, Chezy, Darcy-Weisbach) to calculate flow resistance has been maintained for the canals and deeper waters. A preliminary demonstration exercise WHIMFLO-2D in an existing field site was developed for the Loxahatchee Impoundment Landscape Assessment (LILA), an 80 acre area, located at the Arthur R. Marshall Loxahatchee National Wild Life Refuge in Boynton Beach, Florida. After applying a number of simplifying assumptions, results have illustrated the ability of the model to simulate the hydrology of a wetland. In this illustrative case, a comparison between measured and simulated stages level showed an average error of 0.31% with a maximum error of 2.8%. Comparison of measured and simulated groundwater head levels showed an average error of 0.18% with a maximum of 2.9%. The coupling of FLO-2D model with MODFLOW-2005 model and the incorporation of the dynamic effect of flow resistance due to vegetation performed in the new modeling tool WHIMFLO-2D is an important contribution to the field of numerical modeling of hydrologic flow in wetlands.

Testing the Efficacy of Panax ginseng Extract as a Novel Anti-Quorum Sensing Agent

Quorum sensing (QS) is the phenomenon by which microorganisms regulate gene expression in response to cell-population density. These microorganisms synthesize and secrete small molecules known as autoinducers that increase in concentration as a function of cell density. Once the cell detects the minimal threshold concentration of an autoinducer, the gene expression is altered accordingly. Although the cellular circuitry responding to QS is relatively conserved, the cellular processes regulated by QS, for example are conjugation, motility, sporulation, biofilm formation and production of virulence factors importamt for infection. Since many pathogens have developed resistance to available antibiotics, novel therapies are required to further treat these pathogens. Inhibitors of QS are potential therapeutic candidates since they would inhibit virulence without selecting for antibiotic resistant strains. Previous studies have shown that the Chinese herb Panax ginseng contains compounds that inhibit QS activity. To further characterize this activity, a highly sensitive quantitative liquid assay was developed using a Chromobacterium violaceum AHL mutant, CV026 as a biomonitor strain. After confirmation that P. ginseng aqueous extracts had anti-QS activity, the extract was fractionated on a reverse phase C18 Sep Pak column. Most anti-QS activity was present in the flow through, and compounds eluted with ten percent acetonitrile in water antibacterial activity. We thus conclude that P. ginseng has anti-QS activity and the active compound is water soluble. Compounds from P. ginseng, could be used as a lead structure to design compound with higher anti-QS activity for therapeutic treatments.

Integrated Surface-Ground Water Modeling in Wetlands With Improved Methods to Simulate Vegetative Resistance to Flow

This dissertation focused on developing an integrated surface – subsurface hydrologic simulation numerical model by programming and testing the coupling of the USGS MODFLOW-2005 Groundwater Flow Process (GWF) package (USGS, 2005) with the 2D surface water routing model: FLO-2D (O’Brien et al., 1993). The coupling included the necessary procedures to numerically integrate and verify both models as a single computational software system that will heretofore be referred to as WHIMFLO-2D (Wetlands Hydrology Integrated Model). An improved physical formulation of flow resistance through vegetation in shallow waters based on the concept of drag force was also implemented for the simulations of floodplains, while the use of the classical methods (e.g., Manning, Chezy, Darcy-Weisbach) to calculate flow resistance has been maintained for the canals and deeper waters. A preliminary demonstration exercise WHIMFLO-2D in an existing field site was developed for the Loxahatchee Impoundment Landscape Assessment (LILA), an 80 acre area, located at the Arthur R. Marshall Loxahatchee National Wild Life Refuge in Boynton Beach, Florida. After applying a number of simplifying assumptions, results have illustrated the ability of the model to simulate the hydrology of a wetland. In this illustrative case, a comparison between measured and simulated stages level showed an average error of 0.31% with a maximum error of 2.8%. Comparison of measured and simulated groundwater head levels showed an average error of 0.18% with a maximum of 2.9%.

Design construction and testing of a heat loop for study of fouling and its interrelationship to corrosion in heat exchanger tubes

A heat loop suitable for the study of thermal fouling and its relationship to corrosion processes was designed, constructed and tested. The design adopted was an improvement over those used by such investigators as Hopkins and the Heat Transfer Research Institute in that very low levels of fouling could be detected accurately, the heat transfer surface could be readily removed for examination and the chemistry of the environment could be carefully monitored and controlled. In addition, an indirect method of electrical heating of the heat transfer surface was employed to eliminate magnetic and electric effects which result when direct resistance heating is employed to a test section. The testing of the loop was done using a 316 stainless steel test section and a suspension of ferric oxide and water in an attempt to duplicate the results obtained by Hopkins. Two types of thermal ·fouling resistance versus time curves were obtained . (i) Asymptotic type fouling curve, similar to the fouling behaviour described by Kern and Seaton and other investigators, was the most frequent type of fouling curve obtained. Thermal fouling occurred at a steadily decreasing rate before reaching a final asymptotic value. (ii) If an asymptotically fouled tube was cooled with rapid cir- ·culation for periods up to eight hours at zero heat flux, and heating restarted, fouling recommenced at a high linear rate. The fouling results obtained were observed to be similar and 1n agreement with the fouling behaviour reported previously by Hopkins and it was possible to duplicate quite closely the previous results . This supports the contention of Hopkins that the fouling results obtained were due to a crevice corrosion process and not an artifact of that heat loop which might have caused electrical and magnetic effects influencing the fouling. The effects of Reynolds number and heat flux on the asymptotic fouling resistance have been determined. A single experiment to study the effect of oxygen concentration has been carried out. The ferric oxide concentration for most of the fouling trials was standardized at 2400 ppM and the range of Reynolds number and heat flux for the study was 11000-29500 and 89-121 KW/M², respectively.

Challenges in molecular testing in non-small-cell lung cancer patients with advanced disease

Lung cancer diagnostics have progressed greatly in the previous decade. Development of molecular testing to identify an increasing number of potentially clinically actionable genetic variants, using smaller samples obtained via minimally invasive techniques, is a huge challenge. Tumour heterogeneity and cancer evolution in response to therapy means that repeat biopsies or circulating biomarkers are likely to be increasingly useful to adapt treatment as resistance develops. We highlight some of the current challenges faced in clinical practice for molecular testing of EGFR, ALK, and new biomarkers such as PDL1. Implementation of next generation sequencing platforms for molecular diagnostics in non-small-cell lung cancer is increasingly common, allowing testing of multiple genetic variants from a single sample. The use of next generation sequencing to recruit for molecularly stratified clinical trials is discussed in the context of the UK Stratified Medicine Programme and The UK National Lung Matrix Trial.

Determining phosphine resistance in rust red flour beetle, Tribolium castaneum (Herbst.) (Coleoptera : Tenebrionidae) populations from Turkey

Fumigation with phosphine gas is the primary method of controlling stored grain pests. In Turkey, phosphine has been used extensively since the 1950's. Even though high levels of phosphine resistance have been detected in several key stored products pests across the world, it has never been studied in Turkey despite this long history of phosphine use. High-level phosphine resistance has been detected and genetically characterised previously in the rust red flour beetle, Tribolium castaneum in other countries. Since this pest is also a common problem in stored grain environment in Turkey, the current study was undertaken for the first time, to investigate the distribution and strength of phosphine resistance in T. castaneum. Four strains of T. castaneum were tested through bioassays for determining the weak and strong phosphine resistance phenotypes on the basis of the response of adults to discriminating phosphine concentrations of 0.03 mg/L and 0.25 mg/L, for 20 hour exposures respectively. Phenotype testing showed all strains exhibited some level of phosphine resistance with a maximum level of 196 fold. Sequencing and genetic testing of seven field-collected strains showed that all of them carried a strong resistance allele in at the rph2 locus similar to the one previously reported. Our results show that strong resistance to phosphine is common in Turkish strains of T. castaneum.

Antibiotics resistance of Stenotrophomonas maltophilia strains isolated from various clinical specimens.

Background: A limited number of antibiotics are recommended for the therapy of Stenotrophomonas maltophilia infections due to therapy difficulties caused by its numerous mechanisms of resistance. Objectives: In this study conducted over a period of approximately 5 years we aimed to determine resistance rates of S. maltphilia based on drug classification recommended by Clinical and Laboratory Standards Institute. Methods: A total of 118 S. maltphilia strains isolated from various clinical specimens between January 2006 and June 2012 were included in the study. BD Phoenixautomated microbiology system (Becton Dickinson, USA) was utilized for species level identification and antibiotic susceptibility testing. Results: Sixty seven of S. maltphilia strains were isolated from tracheal aspirate isolates, 17 from blood, 10 from sputum, 10 from wound and 14 from other clinical specimens. Levofloxacin was found to be the most effective antibiotic against S. maltphilia strains with resistance rate of 7.6%. The resistance rates to other antibiotics were as follows: chloramphenicol 18.2%, trimethoprim-sulfamethoxazole 20.3% and ceftazidime 72%. Conclusion: The study revealed that S. maltphilia is resistant to many antibiotics. The treatment of infections caused by S. maltphilia should be preferred primarily as levofloxacin, chloramphenicol, and TMP-SXT, respectively.