Impact of the Human Bacterial Environment on Mycobacteriosis and Allergy

My work describes two sectors of the human bacterial environment: 1. The sources of exposure to infectious non-tuberculous mycobacteria. 2. Bacteria in dust, reflecting the airborne bacterial exposure in environments protecting from or predisposing to allergic disorders. Non-tuberculous mycobacteria (NTM) transmit to humans and animals from the environment. Infection by NTM in Finland has increased during the past decade beyond that by Mycobacterium tuberculosis. Among the farm animals, porcine mycobacteriosis is the predominant NTM disease in Finland. Symptoms of mycobacteriosis are found in 0.34 % of slaughtered pigs. Soil and drinking water are suspected as sources for humans and bedding materials for pigs. To achieve quantitative data on the sources of human and porcine NTM exposure, methods for quantitation of environmental NTM are needed. We developed a quantitative real-time PCR method, utilizing primers targeted at the 16S rRNA gene of the genus of Mycobacterium. With this method, I found in Finnish sphagnum peat, sandy soils and mud high contents of mycobacterial DNA, 106 to 107 genome equivalents per gram. A similar result was obtained by a method based on the Mycobacterium-specific hybridization of 16S rRNA. Since rRNA is found mainly in live cells, this result shows that the DNA detected by qPCR mainly represented live mycobacteria. Next, I investigated the occurrence of environmental mycobacteria in the bedding materials obtained from 5 pig farms with high prevalence (>4 %) of mycobacteriosis. When I used for quantification the same qPCR methods as for the soils, I found that piggery samples contained non-mycobacterial DNA that was amplified in spite of several mismatches with the primers. I therefore improved the qPCR assay by designing Mycobacterium-specific detection probes. Using the probe qPCR assay, I found 105 to 107 genome equivalents of mycobacterial DNA in unused bedding materials and up to 1000 fold more in the bedding collected after use in the piggery. This result shows that there was a source of mycobacteria in the bedding materials purchased by the piggery and that mycobacteria increased in the bedding materials during use in the piggery. Allergic diseases have reached epidemic proportions in urbanized countries. At the same time, childhood in rural environment or simple living conditions appears to protect against allergic disorders. Exposure to immunoreactive microbial components in rural environments seems to prevent allergies. I searched for differences in the bacterial communities of two indoor dusts, an urban house dust shown to possess immunoreactivity of the TH2-type and a farm barn dust with TH1-activity. The immunoreactivities of the dusts were revealed by my collaborators, in vitro in human dendritic cells and in vivo in mouse. The dusts accumulated >10 years in the respiratory zone (>1.5 m above floor), thus reflecting the long-term content of airborne bacteria at the two sites. I investigated these dusts by cloning and sequencing of bacterial 16S rRNA genes from dust contained DNA. From the TH2-active urban house dust, I isolated 139 16S rRNA gene clones. The most prevalent genera among the clones were Corynebacterium (5 species, 34 clones), Streptococcus (8 species, 33 clones), Staphylococcus (5 species, 9 clones) and Finegoldia (1 species, 9 clones). Almost all of these species are known as colonizers of the human skin and oral cavity. Species of Corynebacterium and Streptococcus have been reported to contain anti-inflammatory lipoarabinomannans and immunmoreactive beta-glucans respectively. Streptococcus mitis, found in the urban house dust is known as an inducer of TH2 polarized immunity, characteristic of allergic disorders. I isolated 152 DNA clones from the TH1-active farm barn dust and found species quite different from those found from the urban house dust. Among others, I found DNA clones representing Bacillus licheniformis, Acinetobacter lwoffii and Lactobacillus each of which was recently reported to possess anti-allergy immunoreactivity. Moreover, the farm barn dust contained dramatically higher bacterial diversity than the urban house dust. Exposure to this dust thus stimulated the human dendritic cells by multiple microbial components. Such stimulation was reported to promote TH1 immunity. The biodiversity in dust may thus be connected to its immunoreactivity. Furthermore, the bacterial biomass in the farm barn dust consisted of live intact bacteria mainly. In the urban house dust only ~1 % of the biomass appeared as intact bacteria, as judged by microscoping. Fragmented microbes may possess bioactivity different from that of intact cells. This was recently shown for moulds. If this is also valid for bacteria, the different immunoreactivities of the two dusts may be explained by the intactness of dustborne bacteria. Based on these results, we offer three factors potentially contributing to the polarized immunoreactivities of the two dusts: (i) the species-composition, (ii) the biodiversity and (iii) the intactness of the dustborne bacterial biomass. The risk of childhood atopic diseases is 4-fold lower in the Russian compared with the Finnish Karelia. This difference across the country border is not explainable by different geo-climatic factors or genetic susceptibilities of the two populations. Instead, the explanation must be lifestyle-related. It has already been reported that the microbiological quality of drinking water differs on the two sides of the borders. In collaboration with allergists, I investigated dusts collected from homes in the Russian Karelia and in the Finnish Karelia. I found that bacterial 16S rRNA genes cloned from the Russian Karelian dusts (10 homes, 234 clones) predominantly represented Gram-positive taxa (the phyla Actinobacteria and Firmicutes, 67%). The Russian Karelian dusts contained nine-fold more of muramic acid (60 to 70 ng mg-1) than the Finnish Karelian dusts (3 to 11 ng mg-1). Among the DNA clones isolated from the Finnish side (n=231), Gram-negative taxa (40%) outnumbered the Gram-positives (34%). Out of the 465 DNA clones isolated from the Karelian dusts, 242 were assigned to cultured validly described bacterial species. In Russian Karelia, animal-associated species e.g. Staphylococcus and Macrococcus were numerous (27 clones, 14 unique species). This finding may connect to the difference in the prevalence of allergy, as childhood contacts with pets and farm animals have been connected with low allergy risk. Plant-associated bacteria and plant-borne 16S rRNA genes (chloroplast) were frequent among the DNA clones isolated from the Finnish Karelia, indicating components originating from plants. In conclusion, my work revealed three major differences between the bacterial communtites in the Russian and in the Finnish Karelian homes: (i) the high prevalence of Gram-positive bacteria on the Russian side and of Gram-negative bacteria on the Finnish side and (ii) the rich presence of animal-associated bacteria on the Russian side whereas (iii) plant-associated bacteria prevailed on the Finnish side. One or several of these factors may connect to the differences in the prevalence of allergy.

Analysis of cool roof coatings for residential demand side management in tropical Australia

Cool roof coatings have a beneficial impact on reducing the heat load of a range of building types, resulting in reduced cooling energy loads. This study seeks to understand the extent to which cool roof coatings could be used as a residential demand side management (DSM) strategy for retrofitting existing housing in a constrained network area in tropical Australia where peak electrical demand is heavily influenced by residential cooling loads. In particular this study seeks to determine whether simulation software used for building regulation purposes can provide networks with the ‘impact certainty’ required by their DSM principles. The building simulation method is supported by a field experiment. Both numerical and experimental data confirm reductions in total consumption (kWh) and energy demand (kW). The nature of the regulated simulation software, combined with the diverse nature of residential buildings and their patterns of occupancy, however, mean that simulated results cannot be extrapolated to quantify benefits to a broader distribution network. The study suggests that building data gained from regulatory simulations could be a useful guide for potential impacts of widespread application of cool roof coatings in this region. The practical realization of these positive impacts, however, would require changes to the current business model for the evaluation of DSM strategies. The study provides seven key recommendations that encourage distribution networks to think beyond their infrastructure boundaries, recognising that the broader energy system also includes buildings, appliances and people.

Enzymatic cross-linking of β-casein and its impact on digestibility and allergenicity

Protein modification via enzymatic cross-linking is an attractive way for altering food structure so as to create products with increased quality and nutritional value. These modifications are expected to affect not only the structure and physico-chemical properties of proteins but also their physiological characteristics, such as digestibility in the GI-tract and allergenicity. Protein cross-linking enzymes such as transglutaminases are currently commercially available, but also other types of cross-linking enzymes are being explored intensively. In this study, enzymatic cross-linking of β-casein, the most abundant bovine milk protein, was studied. Enzymatic cross-linking reactions were performed by fungal Trichoderma reesei tyrosinase (TrTyr) and the performance of the enzyme was compared to that of transglutaminase from Streptoverticillium mobaraense (Tgase). Enzymatic cross-linking reactions were followed by different analytical techniques, such as size exclusion chromatography -Ultra violet/Visible multi angle light scattering (SEC-UV/Vis-MALLS), phosphorus nuclear magnetic resonance spectroscopy (31P-NMR), atomic force (AFM) and matrix-assisted laser desorption/ionisation-time of flight mass spectrometry (MALDI-TOF MS). The research results showed that in both cases cross-linking of β-casein resulted in the formation of high molecular mass (MM ca. 1 350 kg mol-1), disk-shaped nanoparticles when the highest enzyme dosage and longest incubation times were used. According to SEC-UV/Vis-MALLS data, commercial β-casein was cross-linked almost completely when TrTyr and Tgase were used as cross-linking enzymes. In the case of TrTyr, high degree of cross-linking was confirmed by 31P-NMR where it was shown that 91 % of the tyrosine side-chains were involved in the cross-linking. The impact of enzymatic cross-linking of β-casein on in vitro digestibility by pepsin was followed by various analytical techniques. The research results demonstrated that enzymatically cross-linked β-casein was stable under the acidic conditions present in the stomach. Furthermore, it was found that cross-linked β-casein was more resistant to pepsin digestion when compared to that of non modified β-casein. The effects of enzymatic cross-linking of β-casein on allergenicity were also studied by different biochemical test methods. On the basis of the research results, enzymatic cross-linking decreased allergenicity of native β-casein by 14 % when cross-linked by TrTyr and by 6 % after treatment by Tgase. It can be concluded that in addition to the basic understanding of the reaction mechanism of TrTyr on protein matrix, the research results obtained in this study can have high impact on various applications like food, cosmetic, medical, textile and packing sectors.

Numerical study of the impact of inoculant and grain transport on macrosegregation and microstructure formation during solidification of an Al-22%Cu alloy

We investigate the impact of the nucleation law for nucleation on Al-Ti-B inoculant particles, of the motion of inoculant particles and of the motion of grains on the predicted macrosegregation and microstructure in a grain-refined Al-22 wt.% Cu alloy casting. We conduct the study by numerical simulations of a casting experiment in a side-cooled 76×76×254 mm sand mould. Macrosegregation and microstructure formation are studied with a volume-averaged two-phase model accounting for macroscopic heat and solute transport, melt convection, and transport of inoculant particles and equiaxed grains. On the microscopic scale it accounts for nucleation on inoculant particles with a given size distribution (and corresponding activation undercooling distribution)and for the growth of globular solid grains. The growth kinetics is described by accounting for limited solute diffusion in both liquid and solid phases and for convective effects. We show that the consideration of a size distribution of the inoculants has a strong impact on the microstructure(final grain size) prediction. The transport of inoculants significantly increases the microstructure heterogeneities and the grain motion refines the microstructure and reduces the microstructure heterogeneities.

Transition from Cassie to impaled state during drop impact on groove-textured solid surfaces

Liquid drops impacted on textured surfaces undergo a transition from the Cassie state characterized by the presence of air pockets inside the roughness valleys below the drop to an impaled state with at least one of the roughness valleys filled with drop liquid. This occurs when the drop impact velocity exceeds a particular value referred to as the critical impact velocity. The present study investigates such a transition process during water drop impact on surfaces textured with unidirectional parallel grooves referred to as groove-textured surfaces. The process of liquid impalement into a groove in the vicinity of drop impact through de-pinning of the three-phase contact line (TPCL) beneath the drop as well as the critical impact velocity were identified experimentally from high speed video recordings of water drop impact on six different groove-textured surfaces made from intrinsically hydrophilic (stainless steel) as well as intrinsically hydrophobic (PDMS and rough aluminum) materials. The surface energy of various 2-D configurations of liquid-vapor interface beneath the drop near the drop impact point was theoretically investigated to identify the locally stable configurations and establish a pathway for the liquid impalement process. A force balance analysis performed on the liquid-vapor interface configuration just prior to TPCL de-pinning provided an expression for the critical drop impact velocity, U-o,U-cr, beyond which the drop state transitions from the Cassie to an impaled state. The theoretical model predicts that Uo, cr increases with the increase in pillar side angle, a, and intrinsic hydrophobicity whereas it decreases with the increase in groove top width, w, of the groove-textured surface. The quantitative predictions of the theoretical model were found to show good agreement with the experimental measurements of U-o,U-cr plotted against the surface texture geometry factor in our model, {tan(alpha/2)/w}(0.5).

Use of side-chain for rational design of n-type diketopyrrolopyrrole-based conjugated polymers: what did we find out?

The primary role of substituted side chains in organic semiconductors is to increase their solubility in common organic solvents. In the recent past, many literature reports have suggested that the side chains play a critical role in molecular packing and strongly impact the charge transport properties of conjugated polymers. In this work, we have investigated the influence of side-chains on the charge transport behavior of a novel class of diketopyrrolopyrrole (DPP) based alternating copolymers. To investigate the role of side-chains, we prepared four diketopyrrolopyrrole-diketopyrrolopyrrole (DPP-DPP) conjugated polymers with varied side-chains and carried out a systematic study of thin film microstructure and charge transport properties in polymer thin-film transistors (PTFTs). Combining results obtained from grazing incidence X-ray diffraction (GIXD) and charge transport properties in PTFTs, we conclude side-chains have a strong influence on molecular packing, thin film microstructure, and the charge carrier mobility of DPP-DPP copolymers. However, the influence of side-chains on optical properties was moderate. The preferential ``edge-on'' packing and dominant n-channel behavior with exceptionally high field-effect electron mobility values of >1 cm(2) V-1 s(-1) were observed by incorporating hydrophilic (triethylene glycol) and hydrophobic side-chains of alternate DPP units. In contrast moderate electron and hole mobilities were observed by incorporation of branched hydrophobic side-chains. This work clearly demonstrates that the subtle balance between hydrophobicity and hydrophilicity induced by side-chains is a powerful strategy to alter the molecular packing and improve the ambipolar charge transport properties in DPP-DPP based conjugated polymers. Theoretical analysis supports the conclusion that the side-chains influence polymer properties through morphology changes, as there is no effect on the electronic properties in the gas phase. The exceptional electron mobility is at least partially a result of the strong intramolecular conjugation of the donor and acceptor as evidenced by the unusually wide conduction band of the polymer.

Investigation of hypervelocity impact phenomena using real-time concurrent diagnostics

Hypervelocity impact of meteoroids and orbital debris poses a serious and growing threat to spacecraft. To study hypervelocity impact phenomena, a comprehensive ensemble of real-time concurrently operated diagnostics has been developed and implemented in the Small Particle Hypervelocity Impact Range (SPHIR) facility. This suite of simultaneously operated instrumentation provides multiple complementary measurements that facilitate the characterization of many impact phenomena in a single experiment. The investigation of hypervelocity impact phenomena described in this work focuses on normal impacts of 1.8 mm nylon 6/6 cylinder projectiles and variable thickness aluminum targets. The SPHIR facility two-stage light-gas gun is capable of routinely launching 5.5 mg nylon impactors to speeds of 5 to 7 km/s. Refinement of legacy SPHIR operation procedures and the investigation of first-stage pressure have improved the velocity performance of the facility, resulting in an increase in average impact velocity of at least 0.57 km/s. Results for the perforation area indicate the considered range of target thicknesses represent multiple regimes describing the non-monotonic scaling of target perforation with decreasing target thickness. The laser side-lighting (LSL) system has been developed to provide ultra-high-speed shadowgraph images of the impact event. This novel optical technique is demonstrated to characterize the propagation velocity and two-dimensional optical density of impact-generated debris clouds. Additionally, a debris capture system is located behind the target during every experiment to provide complementary information regarding the trajectory distribution and penetration depth of individual debris particles. The utilization of a coherent, collimated illumination source in the LSL system facilitates the simultaneous measurement of impact phenomena with near-IR and UV-vis spectrograph systems. Comparison of LSL images to concurrent IR results indicates two distinctly different phenomena. A high-speed, pressure-dependent IR-emitting cloud is observed in experiments to expand at velocities much higher than the debris and ejecta phenomena observed using the LSL system. In double-plate target configurations, this phenomena is observed to interact with the rear-wall several micro-seconds before the subsequent arrival of the debris cloud. Additionally, dimensional analysis presented by Whitham for blast waves is shown to describe the pressure-dependent radial expansion of the observed IR-emitting phenomena. Although this work focuses on a single hypervelocity impact configuration, the diagnostic capabilities and techniques described can be used with a wide variety of impactors, materials, and geometries to investigate any number of engineering and scientific problems.

Impact of hatchery releases on the recreational fishery for Pacific threadfin (Polydactylus sexfilis) in Hawaii

The Pacific threadfin (Polydactylus sexfilis) is considered one of the premier Hawaiian food fishes but even with catch limits, seasonal closures, and size limits, catches have declined dramatically since the 1960s. It was identified as the top candidate species for stock enhancement in Hawaii, based on the decline in stocks, high market value, and importance of the fishery. In the stock enhancement program for Pacific threadfin, over 430,000 fingerlings of various sizes were implanted with coded wire tags and released in nursery habitats along the windward coast of Oahu between 1993 and 1998. Because few Pacific threadfin were present in creel surveys conducted between 1994 and 1998, Oahu fishermen were offered a $10 reward for each threadfin that was caught (for both hatchery-reared and wild fish). A total of 1882 Pacific threadfin were recovered from the reward program between March 1998 and May 1999, including 163 hatchery-reared fish, an overall contribution of 8.7% to the fishery. Hatchery-reared fish accounted for as high as 71% of returns in the release areas. Hatchery-reared fish were recovered on average 11.5 km (SD=9.8 km) from the release site, although some had moved as far away as 42 km. Average age for recovered hatchery-reared fish was 495 days; the oldest was 1021 days. Cultured Pacific threadfin juveniles survived and recruited successfully to the recreational fishery, accounting for 10% of fishermen’s catches on the windward side of Oahu. Recruitment to the fishery was highest for the 1997 release year; few juveniles from earlier releases were observed. Presence of a few large, fully developed females in the recreational fishery suggested that hatchery-reared fish can survive, grow, and reproductively contribute to the population. Implementation of an enhancement program that is focused on juveniles and perhaps large females, as part of an integrated fishery management strategy, could speed the recovery of this fish population.

Coastal development and pollution impact on the distribution of macrobenthic communities along the eastern coast of the Gulf of Suez (Egypt)

The configuration for the eastern side of the Gulf of Suez was studied over 241 km from Ras Mohammed to Ras Sudr including 35 station. Litters, tar balls and aged oil patches aggregated in considerable amounts on the beach and shore line of the middle part due to the oil spills from off-shore oil wells. A large reef flat in the shallow intertidal waters exists at stations 1 and 2 (Ras Mohammed, Protected area) and spars coral patches are less frequent at the Stations from 7 to 13. Density and diversity of marine benthos were higher on hard and cobble bottoms compared to muddy sand and sandy substrates. The assemblages of the benthic fauna are dominated by the gastropod Courmya (Thericium) vulgata; the bivalve Brachiodontes variabilis, and the barnacles Chthamalus stellatus, Balanus amphitrite and Tetraclita rubescens. The distribution of the algal cover in the intertidal region shows high abundance of the brown algae, Sargassum latifolium; padina pavonica and Cystoseira trinodis rather than the green and red algae. These species are found in both polluted and unpolluted areas. The changes in benthic structures in the study area depend not only on the state of pollution but also on the type of substrates.

National report of the frame survey 2012 on the Uganda side of Lake Victoria

This is a report on the results of the Frame Survey conducted in the Uganda side of Lake Victoria during August 2012 by the LVFO Institutions, namely: the Department of Fisheries Resources (DFR) Uganda and the National Fisheries Resources Research Institute (NaFIRRI) in close collaboration with the District Fisheries offices of Busia, Bugiri, Namayingo, Mayuge, Jinja, Buvuma, Buikwe, Mukono, Kampala, Wakiso, Mpigi, Kalungu, Masaka, Kalangala and Rakai. In the 2012 Frame survey some indicators of fishing effort including e.g. number of fishers, fishing crafts and long line hooks increased; whereas others like the number of gillnets less than 5 inches decreased by 10.4% from that recorded in 2010. The other indicators of fishing effort, which showed decrease in 2012 included illegal beach seines and undersized gillnets (<5 inch mesh size). However, a large proportion (66%) of long line hooks recorded in the 2012 survey were in the smallest size range (hook size >10), which target small Nile perch. The number of other illegal gears, i.e. cast nets and monofilament gillnets showed modest increases (25%) between 2010 and 2012 while beach seines decrease by 15%. Recent crackdown on illegal fishing activities as part of measures for recovery of the Nile perch stocks which are faced with depletion appear to have had an impact but much more needs to be done to eradicate illegal fishing. The fisheries in the Ugandan waters have remained predominantly near shore with 61% of all fishing crafts using paddles out of which 17% were tiny three plank, flat bottomed boats locally known as parachutes. The 2012 survey shows an increase in the number of fishing crafts using sails by 65% from 682 in 2010 to 1125 in 2012. This is an encouraging trend as more fishers are able to access distant fishing grounds using free wind power. The Mukene fishery in the Ugandan waters of Lake Victoria remained underdeveloped comprising only 15.2% of all fishing crafts, of which 31% were motorised which is a great improvement from the situation recorded in 2010. The Catamarans increased to 18 with a majority in Buikwe district where there is a private investor fishing specifically for Mukene. The Catamarans in Kalangala were reported not to be working because of the high operating cost compared to ordinary Mukene fishing boats.

Moderate speed impact damage to 2D-braided glass-carbon composites

Hybrid glass-carbon 2D braided composites with varying carbon contents are impacted using a gas gun by impactors of masses 12.5 and 44.5. g, at impact energies up to 50. J. The damage area detected by ultrasound C-scan is found to increase roughly linearly with impact energy, and is larger for the lighter impactor at the same impact energy. The area of whitening of the glass tows on the distal side corresponds with the measured C-scan damage area. X-ray imaging shows more intense damage, at the same impact energy, for a higher-mass impactor. Braids with more glass content have a modest increase in density, decrease in modulus, and reduction in the C-scan area and dent depth at the impact site, particularly at the higher impact energies. Impact damage is found to reduce significantly the compressive strength, giving up to a 26% reduction at the maximum impact energy. © 2012 Elsevier Ltd.

The impact of chemotherapy dose density and dose intensity on breast cancer outcome: what have we learned?

Optimising chemotherapy dose density and dose intensity are strategies aimed at improving outcomes in adjuvant therapy for patients with breast cancer. There are, in theory, at least five models allowing the delivery of a higher overall drug dose intensity. These are reviewed in this article and vary according to three main variables: the dose per course, the interval between doses and the total cumulative dose. Cyclophosphamide, anthracyclines and taxanes are among the most active agents for the treatment of breast cancer and, as such, they have been or are currently the focus of prospective, randomised clinical trials testing some of these dose-intensity models in the adjuvant setting. The results of recent trials suggest that anthracyclines, but not cyclophosphamide, are associated with better outcomes if used at higher doses per course and at higher cumulative doses. However, care has to be taken with premenopausal women where an increased dose of anthracycline per course but a reduced cumulative dose appears to produce a worse outcome. Moreover, decreasing the interval between doses, for anthracyclines and cyclophosphamide, does not seem to provide, so far, additional benefits for women with locally advanced breast cancer. This approach is not feasible with docetaxel, since an increase in dose density induces unwanted side-effects. These results represent our current state of knowledge, but clinical trials are being performed to evaluate further the effect of dose intensity, dose density and cumulative dose of key therapeutic agents on patient outcomes.

Investigating the impact of exit availability on egress time using computer based evacuation simulation

This paper examines the influence of exit availability on evacuation time for a narrow body aircraft under certification trial conditions using computer simulation. A narrow body aircraft which has previously passed the certification trial is used as the test configuration. While maintaining the certification requirement of 50% of the available exits, six different exit configurations are examined. These include the standard certification configuration (one exit from each exit pair) and five other exit configurations based on commonly occurring exit combinations found in accidents. These configurations are based on data derived from the AASK database and the evacuation simulations are performed using the airEXODUS evacuation simulation software. The results show that the certification practice of using half the available exits predominately down one side of the aircraft is neither statistically relevant nor challenging. For the aircraft cabin layout examined, the exit configuration used in certification trial produces the shortest egress times. Furthermore, three of the six exit combinations investigated result in predicted egress times in excess of 90 seconds, suggesting that the aircraft would not satisfy the certification requirement under these conditions.

Investigating the impact of aircraft exit availability on egress time using computer simulation

This paper examines the influence of exit availability on evacuation time for narrow body aircraft under certification trial conditions using computer simulation. A narrow body aircraft which has previously passed the certification trial is used as the test configuration. While maintaining the certification requirement of 50% of the available exits, six different configurations are examined. These include the standard certification and five other exit configurations based on commonly occurring exit combinations found in accidents. These configurations are based on data derived from the AASK database and the evacuation simulations are performed using the airEXODUS evacuation software. The results show that the certification practise of using half of the available exits predominately down one side of the aircraft is neither statistically relevant nor challenging. For the aircraft cabin layout examined, the exit configuration used in certification trial produces the shortest egress times. Furthermore, three of the six exit combinations investigated result in predicted egress times in excess of 90 seconds, suggesting that the aircraft would not satisfy the certification requirement under these conditions.

Financing of SME's: an asset side story

The main sources of financing for small and medium sized enterprises (SMEs) are equity (internally generated cash), trade credit paid on time, long and short term bank credits, delayed payment on trade credit and other debt. The marginal costs of each financing instrument are driven by asymmetric information (cost of gathering and analysing information) and transactions costs associated with non-payment (costs of collecting and selling collateral). According to the Pecking Order Theory, firms will choose the cheapest source in terms of cost. In the case of the static trade-off theory, firms choose finance so that the marginal costs across financing sources are all equal, thus an additional Euro of financing is obtained from all the sources whereas under the Pecking Order Theory the source is determined by how far down the Pecking Order the firm is presently located. In this paper, we argue that both of these theories miss the point that the marginal costs are dependent of the use of the funds, and the asset side of the balance sheet primarily determines the financing source for an additional Euro. An empirical analysis on a unique dataset of Portuguese SME's confirms that the composition of the asset side of the balance sheet has an impact of the type of financing used and the Pecking Order Theory and the traditional Static Trade-off theory are rejected.