The potential of microbial ecological indicators to guide ecosophisticated management of hydrocarbon-contaminated soils

Soil is an unrenewable natural resource under increasing anthropogenic pressure. One of the main threats to soils, compromising their ability to provide us with the goods and ecosystem services we expect, is pollution. Oil hydrocarbons are the most common soil contaminants, and they disturb not just the biota but also the physicochemical properties of soils. Indigenous soil micro-organisms respond rapidly to changes in the soil ecosystem, and are chronically in direct contact with the hydrophobic pollutants on the soil surfaces. Soil microbial variables could thus serve as an intrinsically relevant indicator of soil quality, to be used in the ecological risk assessment of contaminated and remediated soils. Two contrasting studies were designed to investigate soil microbial ecological responses to hydrocarbons, together with parallel changes in soil physicochemical and ecotoxicological properties. The aim was to identify quantitative or qualitative microbiological variables that would be practicable and broadly applicable for the assessment of the quality and restoration of oil-polluted soil. Soil bacteria commonly react on hydrocarbons as a beneficial substrate, which lead to a positive response in the classical microbiological soil quality indicators; negative impacts were accurately reflected only after severe contamination. Hydrocarbon contaminants become less bioavailable due to weathering processes, and their potentially toxic effects decrease faster than the total concentration. Indigenous hydrocarbon degrader bacteria, naturally present in any terrestrial environment, use specific mechanisms to improve access to the hydrocarbon molecules adsorbed on soil surfaces. Thus when contaminants are unavailable even to the specialised degraders, they should pose no hazard to other biota either. Change in the ratio of hydrocarbon degrader numbers to total microbes was detected to predictably indicate pollutant effects and bioavailability. Also bacterial diversity, a qualitative community characteristic, decreased as a response to hydrocarbons. Stabilisation of community evenness, and community structure that reflected clean reference soil, indicated community recovery. If long-term temporal monitoring is difficult and appropriate clean reference soil unavailable, such comparison could possibly be based on DNA-based community analysis, reflecting past+present, and RNA-based community analysis, showing exclusively present conditions. Microbial ecological indicators cannot replace chemical oil analyses, but they are theoretically relevant and operationally practicable additional tools for ecological risk assessment. As such, they can guide ecologically informed and sustainable ecosophisticated management of oil-contaminated lands.

Kelan sotilasavustustilasto 2010

Suomen virallinen tilasto

Moonlighting Proteins of Lactobacillus crispatus : Extracellular Localization, Cell Wall Anchoring and Interactions with the Host

Moonlighting functions have been described for several proteins previously thought to localize exclusively in the cytoplasm of bacterial or eukaryotic cells. Moonlighting proteins usually perform conserved functions, e. g. in glycolysis or as chaperonins, and their traditional and moonlighting function(s) usually localize to different cell compartments. The most characterized moonlighting proteins in Grampositive bacteria are the glycolytic enzymes enolase and glyceraldehyde-3-phosphate dehydrogenase (GAPDH), which function in bacteria-host interactions, e. g. as adhesins or plasminogen receptors. Research on bacterial moonlighting proteins has focused on Gram-positive bacterial pathogens, where many of their functions have been associated with bacterial virulence. In this thesis work I show that also species of the genus Lactobacillus have moonlighting proteins that carry out functions earlier associated with bacterial virulence only. I identified enolase, GAPDH, glutamine synthetase (GS), and glucose-6-phosphate isomerase (GPI) as moonlighting proteins of Lactobacillus crispatus strain ST1 and demonstrated that they are associated with cell surface and easily released from the cell surface into incubation buffer. I also showed that these lactobacillar proteins moonlight either as adhesins with affinity for basement membrane and extracellular matrix proteins or as plasminogen receptors. The mechanisms of surface translocation and anchoring of bacterial moonlighting proteins have remained enigmatic. In this work, the surface localization of enolase, GAPDH, GS and GPI was shown to depend on environmental factors. The members of the genus Lactobacillus are fermentative organisms that lower the ambient pH by producing lactic acid. At acidic pH enolase, GAPDH, GS and GPI were associated with the cell surface, whereas at neutral pH they were released into the buffer. The release did not involve de novo protein synthesis. I showed that purified recombinant His6-enolase, His6-GAPDH, His6-GS and His6-GPI reassociate with cell wall and bind in vitro to lipoteichoic acids at acidic pH. The in-vitro binding of these proteins localizes to cell division septa and cell poles. I also show that the release of moonlighting proteins is enhanced in the presence of cathelicidin LL- 37, which is an antimicrobial peptide and a central part of the innate immunity defence. I found that the LL-37-induced detachment of moonlighting proteins from cell surface is associated with cell wall permeabilization by LL-37. The results in this thesis work are compatible with the hypothesis that the moonlighting proteins of L. crispatus associate to the cell wall via electrostatic or ionic interactions and that they are released into surroundings in stress conditions. Their surface translocation is, at least in part, a result from their release from dead or permeabilized cells and subsequent reassociation onto the cell wall. The results of this thesis show that lactobacillar cells rapidly change their surface architecture in response to environmental factors and that these changes influence bacterial interactions with the host.

The function and regulation of the U11-48K protein in U12-dependent splicing

The removal of noncoding sequences, or introns, from the eukaryotic messenger RNA precursors is catalyzed by a ribonucleoprotein complex known as the spliceosome. In most eukaryotes, two distinct classes of introns exist, each removed by a specific type of spliceosome. The major, U2-type introns account for over 99 % of all introns, and are almost ubiquitous. The minor, U12-type introns are found in most but not all eukaryotes, and reside in conserved locations in a specific set of genes. Due to their slow excision rates, the U12-type introns are expected to be involved in the regulation of the genes containing them by inhibiting the maturation of the messenger RNAs. However, little information is currently available on how the activity of the U12-dependent spliceosome itself is regulated. The levels of many known splicing factors are regulated through unproductive alternative splicing events, which lead to inclusion of premature STOP codons, targeting the transcripts for destruction by the nonsense-mediated decay pathway. These alternative splice sites are typically found in highly conserved sequence elements, which also contain binding sites for factors regulating the activation of the splice sites. Often, the activation is achieved by binding of products of the gene in question, resulting in negative feedback loops. In this study, I show that U11-48K, a protein factor specific to the minor spliceosome, specifically recognizes the U12-type 5' splice site sequence, and is essential for proper function of the minor spliceosome. Furthermore, the expression of U11-48K is regulated through a feedback mechanism, which functions through conserved sequence elements that activate alternative splicing and nonsense-mediated decay. This mechanism is conserved from plants to animals, highlighting both the importance and early origin of this mechanism in regulating splicing factors. I also show that the feedback regulation of U11-48K is counteracted by a component of the major spliceosome, the U1 small nuclear ribonucleoprotein particle, as well as members of the hnRNP F/H protein family. These results thus suggest that the feedback mechanism is finely tuned by multiple factors to achieve precise control of the activity of the U12-dependent spliceosome.

Role of GDNF and its Cross-Talk with Other Growth Factors in the Dopaminergic System

Parkinson´s Disease (PD) is a neurodegenerative movement disorder resulting from loss of dopaminergic (DA) neurons in substantia nigra (SN). Possible causative treatment strategies for PD include neurotrophic factors, which protect and in some cases restore the function of dopaminergic neurons. Glial cell line-derived neurotrophic factor (GDNF) family of neurotrophic factors have been to date the most promising candidates for treatment of PD, demonstrating both neuroprotective and neurorestorative properties. We have investigated the role of GDNF in the rodent dopaminergic system and its possible crosstalk with other growth factors. We characterized the GDNF-induced gene expression changes by DNA microarray analysis in different neuronal systems, including in vitro cultured Neuro2A cells treated with GDNF, as well as midbrains from GDNF heterozygous (Hz) knockout mice. These microarray experiments, resulted in the identification of GDNF-induced genes, which were also confirmed by other methods. Further analysis of the dopaminergic system of GDNF Hz mice demonstrated about 40% reduction in GDNF levels, revealed increased intracellular dopamine concentrations and FosB/DeltaFosB expression in striatal areas. These animals did not show any significant changes in behavioural analysis of acute and repeated cocaine administration on locomotor activity, nor did they exhibit any changes in dopamine output following treatment with acute cocaine. We further analysed the significance of GDNF receptor RET signalling in dopaminergic system of MEN2B knock-in animals with constitutively active Ret. The MEN2B animals showed a robust increase in extracellular dopamine and its metabolite levels in striatum, increased tyrosine hydroxylase (TH) and dopamine transporter (DAT) protein levels by immunohistochemical staining and Western blotting, as well as increased Th mRNA levels in SN. MEN2B mice had increased number of DA neurons in SN by about 25% and they also exhibited increased sensitivity to the stimulatory effects of cocaine. We also developed a semi-throughput in vitro micro-island assay for the quantification of neuronal survival and TH levels by computer-assisted methodology from limited amounts of tissue. This assay can be applied for the initial screening for dopaminotrophic molecules, as well as chemical drug library screening. It is applicable to any neuronal system for the screening of neurotrophic molecules. Since our microarray experiments revealed possible GDNF-VEGF-C crosstalk we further concentrated on studying the neurotrophic effects of VEGF-C. We showed that VEGF-C acts as a neurotrophic molecule for the DA neurons both in vitro and in vivo, however without additive effect when used together with GDNF. The neuroprotective effect for VEGF-C in vivo in rat 6-OHDA model of PD was demonstrated. The possible signalling mechanisms of VEGF-C in the nervous system were investigated - infusion of VEGF-C to rat brain induced ERK activation, however no direct activation of RET signalling in vitro was found. VEGF-C treatment of rat striatum lead to up-regulation of VEGFR-1-3, indicating that VEGF-C can regulate the expression level of its own receptor. VEGF-C dopaminotrophic activity in vivo was further supported by increased vascular tissue in the neuroprotection experiments.

Biocompatibility and biofunctionalization of mesoporous silicon particles

Several of the newly developed drug molecules experience poor biopharmaceutical behavior, which hinders their effective delivery at the proper site of action. Among the several strategies employed in order to overcome this obstacle, mesoporous silicon-based materials have emerged as promising drug carriers due to their ability to improve the dissolution behavior of several poorly water-soluble drugs compounds confined within their pores. In addition to improve the dissolution behavior of the drugs, we report that porous silicon (PSi) nanoparticles have a higher degree of biocompatibility than PSi microparticles in several cell lines studied. In addition, the degradation of the nanoparticles showed its potential to fast clearance in the body. After oral delivery, the PSi particles were also found to transit the intestines without being absorbed. These results constituted the first quantitative analysis of the behavior of orally administered PSi nanoparticles compared with other delivery routes in rats. The self-assemble of a hydrophobin class II (HFBII) protein at the surface of hydrophobic PSi particles endowed the particles with greater biocompatibility in different cell lines, was found to reverse their hydrophobicity and also protected a drug loaded within its pores against premature release at low pH while enabling subsequent drug release as the pH increased. These results highlight the potential of HFBII-coating for PSi-based drug carriers in improving their hydrophilicity, biocompatibility and pH responsiveness in drug delivery applications. In conclusion, mesoporous silicon particles have been shown to be a versatile platform for improving the dissolution behavior of poorly water-soluble drugs with high biocompatibility and easy surface modification. The results of this study also provide information regarding the biofunctionalization of the THCPSi particles with a fungal protein, leading to an improvement in their biocompatibility and endowing them with pH responsive and mucoadhesive properties.

Importance of COMT in the regulation of prefrontal dopamine

The prefrontal cortex (PFC), located in the anterior region of the frontal lobe, is considered to have several key roles in higher cognitive and executive functions. In general, the PFC can be seen as a coordinator of thought and action allowing subjects to behave in a goal-directed manner. Due to its anatomical connections with a variety of cortical and subcortical structures, several neurotransmitters, including dopamine, are involved in the regulation of PFC activity. In general, the majority of released dopamine is cleared by the dopamine transporter (DAT). In the PFC however, the number of presynaptic DAT is diminished, emphasizing the relative importance of catechol-O-methyltransferase (COMT) in dopamine metabolism. As a result, the role of COMT in the etiology of psychotic disorders is under constant debate. The present study investigated the role of COMT in prefrontal cortical dopamine metabolism by different neurochemical methods in COMT knockout (COMT-KO) mice. Pharmacological tools to inhibit other dopamine clearing mechanisms were also used for a more comprehensive and collective picture. In addition, this study investigated how a lack of the soluble (S-) COMT isoform affects the total COMT activity as well as the pharmacokinetics of orally administered L-dopa using mutant mice expressing only the membrane-bound (MB-) COMT isoform. Also the role of COMT in striatal and accumbal dopamine turnover during Δ9-tetrahydrocannabinol (THC) challenge was studied. We found markedly increased basal dopamine concentrations in the PFC, but not the striatum or nucleus accumbens (NAcc), of mice lacking COMT. Pharmacological inhibition of the noradrenaline transporter (NET) and monoamine oxidase (MAO) elevated prefrontal cortical dopamine levels several-fold, whereas inhibition of DAT did not. The lack of COMT doubled the dopamine raising effects of NET and MAO inhibition. No compensatory expression of either DAT or NET was found in the COMT-KO mice. The lack of S-COMT decreased the total COMT activity by 50-70 % and modified dopamine transmission and the pharmacokinetics of exogenous Ldopa in a sex and tissue specific manner. Finally, we found that subsequent tolcapone and THC increased dopamine levels in the NAcc, but not in the striatum. Conclusively, this study presents neurochemical evidence for the important role of COMT in the PFC and shows that COMT is responsible for about half of prefrontal cortical dopamine metabolism. This study also highlights the previously underestimated proportional role of MB-COMT and supports the clinical evidence of a gene x environment interaction between COMT and cannabis.

Young Actors in Transnational Agoras : Multi-sited Ethnography of Cosmopolitan Micropolitical Orientations

This study examines young people s political participation in transnational meetings. Methodologically the study aims to shed light on multi-sited global ethnography. Young people are viewed here as a social age group sensitive to critical, alternative and even radical political participation. The diversity of the young actors and their actions is captured by using several different methods. What is more, the study spurs us coming from the Global North to develop social science research towards methodological cosmopolitanism and to consider our research practices from a moral cosmopolitan perspective. The research sites are the EU Presidency Youth Event (2006 Hyvinkää, Finland), the Global Young Greens Founding Conference (2007 Nairobi, Kenya), the European Social Forum (2008 Malmö, Sweden) and three World Social Forums (2006 Bamako, Mali; 2007 Nairobi Kenya and 2009 Belém, Brazil). The data consists of participant observation, documents and media articles of the meetings, interviews, photos, video, and internet data. This multidisciplinary study combines youth research, development studies, performative social science and political sociology. In this research the diverse field of youth political participation in transnational agoras is studied by using a cross-table of cosmopolitan resources (or the lack of them) and everydaymakers expert citizen dichotomy. First, the young participants of the EU Presidency youth event are studied as an example of expert citizens with cosmopolitan resources (these resources include, for example, language skills, higher education and international social network). Second, the study analyses those everyday-makers who use performative politics to demonstrate their political missions here and now. But in order to make the social movement global they need cosmopolitan resources to be able to use the social media tools and work globally. Third, the study reflects upon the difficulties of reaching those actors who lack cosmopolitan resources, either everyday-makers or expert citizens. The go-along method and the use of the interpreters are shown as ways to reach these young people s political missions. Fourth, the research underlines the importance of contact zones (i.e. spaces or situations where the aforementioned orientations and their differences temporarily disappear or weaken) for deeper democracy and for boosted dialogue between different kinds of participants. Keywords: political participation, young people, multi-sited ethnography, youth research, political sociology, development studies, performative social science

Performability of automated manufacturing systems with centralized material handling

We present a framework for performance evaluation of manufacturing systems subject to failure and repair. In particular, we determine the mean and variance of accumulated production over a specified time frame and show the usefulness of these results in system design and in evaluating operational policies for manufacturing systems. We extend this analysis for lead time as well. A detailed performability study is carried out for the generic model of a manufacturing system with centralized material handling. Several numerical results are presented, and the relevance of performability analysis in resolving system design issues is highlighted. Specific problems addressed include computing the distribution of total production over a shift period, determining the shift length necessary to deliver a given production target with a desired probability, and obtaining the distribution of Manufacturing Lead Time, all in the face of potential subsystem failures.

Ultraviolet photoelectron spectroscopy of the hydrogen bonded heterodimer H2Scdots, three dots, centeredHCl

The HeI photoelectron spectrum of the hydrogen bonded hetero-dimer H2Scdots, three dots, centeredHCl shows two vertical ionization energies at 10.91 and 12.16 eV. Ab initio MO calculations reveal that these features are due to the sulphur and chlorine lone pair ionizations respectively. Results show that while the ground ionic state is repulsive the first excited ionic state is strongly bound. The photoelectron spectrum of the diethyl sulphidecdots, three dots, centeredHCl complex is similar to that of H2Scdots, three dots, centeredHCl.

Polyethylene stibinite phosphate esters: Novel flame-retardant plasticizers for PVC

A new series of multielement flame-retardant plasticizers containing polyethylene stibinite phosphate esters have been prepared by bulk polymerization from ethylene glycol with various antimony (III) aryloxydichlorides and arylphosphorodichloridates possessing various combinations of substituent [Cl,Br,NO2]. All the polymers are pink-coloured viscous fluids. They were characterized by inherent viscosity, density, IR, H-1, C-13 and P-31 NMR spectroscopy. The thermal behaviour of the polymers was compared by thermogravimetric analysis and correlated with their structures. The flammability studies were carried out by the limiting oxygen index test. The polymers containing P, Sb, N and Pr elements in their backbone show superior thermal-and flame-retardant characteristics than the other polymers. A comparative study was carried out with one of the synthesized polymers as a polymeric flame-retardant additive to plasticized PVC. The results showed improved LOI and mechanical properties to that of the conventional flame-retardant additive composition. (C) 1997 Elsevier Science Ltd.

Squeeze-film Effects in MEMS Devices with Perforated Plates for Small Amplitude Vibration

Squeeze-film effects of perforated plates for small amplitude vibration are analyzed through modified Reynolds equation (MRE). The analytical analysis reckons in most important influential factors: compressibility of the air, border effects, and the resistance caused by vertical air flow passing through perforated holes. It is found that consideration of air compressibility is necessary for high operating frequency and small ratio of the plate width to the attenuation length. The analytical results presented in this paper agree with ANSYS simulation results better than that under the air incompressibility assumption. The analytical analysis can be used to estimate the squeeze-film effects causing damping and stiffness added to the system. Since the value of Reynolds number involved in this paper is low (< 1), inertial effects are neglected.

Comportamiento agronómico de dos cultivares clonales de quequisque (Xanthosoma sagittifolium (L) schott), en condiciones de Yolaina, Nueva Guinea, primera 2000-2001

Con el objetivo de caracterizar agronómicamente dos genotipos de quequisque (Xanthosoma sagittiolium (L) Schott), se estableció un ensayo en condiciones de campo de Yolaina, municipio de Nueva Guinea, en época de primera del 7 junio del 2000 al 23 de marzo del 2001 se evaluó las variables morfológicas, fenológicas, de rendimiento y la presencia de enfermedades (virus) en los cultivares Masaya (My) y Nueva Guinea (NG). Se utilizó un diseño de bloque completos al azar (BCA), con 4 bloques y 2 tratamiento por bloque. La parcela experimental estuvo conformada por 6 surcos, con una longitud de 12 m. La distancia de siembra de 1m entre surco y 0.60 m entre planta para un total de 20 plantas por surco y 240 plantas por parcela, para un total de 960 plantas en el ensayo. El área total del experimento fue de 729m2, el área de tratamiento de 144m2 y los dos surcos centrales representaron la parcela útil, sin incluir las primeras 5 plantas en ambos extremos. Las variables evaluadas fueron: altura de planta (cm), área foliar (cm 2), número de hojas, número de hijos y grosor de pseudotallo (cm), de rendimiento: número de cormelos por planta, peso de cormelos totales por planta (g), peso promedio de cormelo (g), largo de cormelo (cm) y diámetro de cormelo (cm). Los eventos morfológicos velocidad de brotación y momento de cosecha: presencia de enfermedades (DMV). A las medias de los datos se les realizó un ANDEVA y separación de medias con la prueba de rangos múltiples de Duncan. Indicando que los cultivares presentaron diferencias estadísticas entre ellas en las variables morfológicas habiendo obtenido el genotipo My los mayores valores en: altura de planta con (70.3 cm), área foliar con (1,540 cm2), número de hojas con (3.79), grosor de pseudotallo por planta (1,540 cm) y para el número de brotes el genotipo NG obtuvo los mayores valores con (0.172). En los componentes de rendimiento el genotipo NG reportó los mayores valores: número de cormelo por planta (3.84), peso de cormelo por planta (464.8 g), largo de cormelo (12.34 cm) y el genotipo My reportó los mayores valores en diámetro de cormelo con (5.19 cm) y peso promedio (130.5 g). Se realizaron 3 conteos visuales de la presencia del virus. Se encontró tendencia a aumentar y disminuir el porcentaje de plantas con presencia del DMV con el aumento de los días después de la siembra. El genotipo My reportó un máximo de (27.7%) y NG (25.3%) a los 76 dds. En los eventos fenológicos la velocidad de brotación y la traslocación temprana de nutrientes (número de hojas (3.56), reducción del área foliar (1,085) y la presencia de cormelos con las yemas apicales y axilares brotadas (41.73%) y con presencia de raíces en crecimiento con (78.26%) al momento de la cosecha, con esto se demostró que el cultivar NG fue más precoz que el My.