Benzene and toluene are representative volatile organic compounds (VOCs) released during production, storage, and transportation associated with the oil and gas industry. Benzene and toluene are chemicals of concern because they are released in greater and possibly more biologically significant concentrations than other compounds. <p>Most studies of air pollution in high oil and gas activity areas have neglected to consider risks to top-level predators. Birds can be used as highly sensitive monitors of air quality. Since the avian respiratory tract is physiologically different from a rodent respiratory tract, effects of gases cannot be safely extrapolated from rodent studies. I hypothesized that benzene, being haematotoxic and immunotoxic, along with the neurological and possible endocrine disrupting effects of toluene would be more toxic in birds than in mammals. <p>After two summers of experimental exposure of wild and captive American kestrels to high (10ppm and 80ppm) or environmentally relevant (0.1ppm and 0.8ppm) levels of benzene and toluene, respectively, altered immune, haematopoeitic, behavioural, and endocrine responses characteristic in mammals, were evident in the kestrels.<p>There was a decreased cell mediated immune response as measured by delayed type hypersensitivity tests in all exposed birds (p = 0.028, 0.004). An increase in humoral immunity as compared to control individuals (p = 0.041, 0.031) was also apparent in both dose groups. Plasma retinol levels were decreased in 2005 and 2006 high dose individuals (p = 0.008, 0.048). <p>The majority of haematopoeitic effects involved the erythroid lineage in the bone marrow and the polychromatophilic erythrocytes systemically. There were no significantly adverse responses in the bone marrow with regards to the granuloid lineage but systemically there was a prominent eosinophilia (p = 0.045) and basophilia (p = 0.006) in low exposure groups. The loss of communication between polychromatophilic erythrocytes in the post-mitotic pool within the bone marrow and the peripheral blood was present in low and high exposure individuals compared to control birds (p = 0.013, 0.402, 0.974). The number of polychromatophils in the circulation of low dose group individuals was decreased compared to control birds (p = 0.029). This may be a function of toluenes inability to inhibit biotransformation enzymes at low concentrations leading to blood cell targeting by benzenes increased phenolic metabolite production. This theory is corroborated by the possible decreased benzene metabolism and increased toluene distribution manifesting as increased aggressive responses such as wing beating and vocalization time in the high dose group (p = 0.025, 0.086). <p>The work here has shown American kestrels are sensitive to the air contaminants, benzene and toluene. The present study illustrates the need for reference concentrations for airborne pollutants that are calculated based on data measuring sensitive endpoints specific for avian models. Future studies should evaluate immune, haematopoeitic, and behavioural endpoints, as well as develop more sensitive isoform specific enzyme activity assays to further determine the susceptibility of birds to inhaled toxicants.

L'objectiu d'aquest estudi és el d'investigar sobre l'ús de matèria orgànica per part dels fongs i bacteris que colonitzen diferents substrats bentònics en rius Mediterranis i analitzar l'efecte dels factors ambientals i antròpics sobre l'estabilitat estructural i funcional de les comunitats del biofilm. La metodologia emprada en aquest estudi consisteix en: i) anàlisi de la biomassa bacteriana i fúngica, ii) anàlisi de la composició de les comunitats bentòniques (identificació d'hifomicets aquàtics i anàlisi del 16S rDNA bacterià), i iii) anàlisi de l'activitat enzimàtica extracel·lular relacionada amb el reciclatge de matèria orgànica en rius. / This study aimed to investigate on the use of organic matter by fungi and bacteria inhabiting different benthic substrata and to analyze the effect of environmental and anthropogenic perturbations on the structural and functional stability of biofilms. The following methodologies has been used in this study: i) analysis of fungal and bacterial biomass, ii) analysis of benthic community composition (identification of hyphomycete taxa, analysis of the bacterial 16S rDNA), and iii) analysis of extracellular enzyme activities involved in organic matter recycling in rivers.

Benzene and toluene are representative volatile organic compounds (VOCs) released during production, storage, and transportation associated with the oil and gas industry. Benzene and toluene are chemicals of concern because they are released in greater and possibly more biologically significant concentrations than other compounds. <p>Most studies of air pollution in high oil and gas activity areas have neglected to consider risks to top-level predators. Birds can be used as highly sensitive monitors of air quality. Since the avian respiratory tract is physiologically different from a rodent respiratory tract, effects of gases cannot be safely extrapolated from rodent studies. I hypothesized that benzene, being haematotoxic and immunotoxic, along with the neurological and possible endocrine disrupting effects of toluene would be more toxic in birds than in mammals. <p>After two summers of experimental exposure of wild and captive American kestrels to high (10ppm and 80ppm) or environmentally relevant (0.1ppm and 0.8ppm) levels of benzene and toluene, respectively, altered immune, haematopoeitic, behavioural, and endocrine responses characteristic in mammals, were evident in the kestrels.<p>There was a decreased cell mediated immune response as measured by delayed type hypersensitivity tests in all exposed birds (p = 0.028, 0.004). An increase in humoral immunity as compared to control individuals (p = 0.041, 0.031) was also apparent in both dose groups. Plasma retinol levels were decreased in 2005 and 2006 high dose individuals (p = 0.008, 0.048). <p>The majority of haematopoeitic effects involved the erythroid lineage in the bone marrow and the polychromatophilic erythrocytes systemically. There were no significantly adverse responses in the bone marrow with regards to the granuloid lineage but systemically there was a prominent eosinophilia (p = 0.045) and basophilia (p = 0.006) in low exposure groups. The loss of communication between polychromatophilic erythrocytes in the post-mitotic pool within the bone marrow and the peripheral blood was present in low and high exposure individuals compared to control birds (p = 0.013, 0.402, 0.974). The number of polychromatophils in the circulation of low dose group individuals was decreased compared to control birds (p = 0.029). This may be a function of toluenes inability to inhibit biotransformation enzymes at low concentrations leading to blood cell targeting by benzenes increased phenolic metabolite production. This theory is corroborated by the possible decreased benzene metabolism and increased toluene distribution manifesting as increased aggressive responses such as wing beating and vocalization time in the high dose group (p = 0.025, 0.086). <p>The work here has shown American kestrels are sensitive to the air contaminants, benzene and toluene. The present study illustrates the need for reference concentrations for airborne pollutants that are calculated based on data measuring sensitive endpoints specific for avian models. Future studies should evaluate immune, haematopoeitic, and behavioural endpoints, as well as develop more sensitive isoform specific enzyme activity assays to further determine the susceptibility of birds to inhaled toxicants.

Phosphoglucomutase (PGM) is one of the central enzymes in energy metabolism at a branch point at the head of the metabolic pathway leading into glycogen metabolism, pentose shunt and the main glycolytic cycle, catalyzing the reversible interconversion of glucose-1-phosphate to glucose-6-phosphate. Whole year, month to month analysis of pattern of allozyme variation at Pgm and Hk loci in Apis mellifera L. from three provinces / Kirklareli, Artvin, and Hatay revealed that there is significant seasonal variation of allozyme frequencies at Pgm locus (P&lt / 0.001). The difference in genotype frequencies between summer and winter samples is apparent in Pgm, whereas at Hk locus, which is analyzed as a control there is seasonal variation in genotype frequencies. Biochemical measurements of the enzyme activities and glycogen content of different Pgm genotypes were performed to determine the effect of different Pgm genotypes on the physiological performance of the honeybees and it was observed that both enzyme activity and glycogen amount is higher in heterozygote individuals which are in high frequency during winter months (P&lt / 0.0001). Furthermore, PGM enzyme activity and glycogen content was found to be significantly correlated. These findings clearly demonstrate that biochemical differences between different Pgm genotypes have functional correlates that lead to significant variations in glycogen content of the honeybees and may have adaptive consequences.

Mittels immunhistochemischer- und zyto-fotometrischer Verfahren wurden die Änderungen der glykolytischen und der oxidativen Enzymkapazität im Skelettmuskel von Patienten mit T2DM mit der Muskelfasercharakteristik untersucht. Weiterführende Analysen klären den Zusammenhang zwischen der Änderung der Muskelfaserverteilung und der Enzymkapazität. Durch eine perkutane Biopsie des M. vastus lateralis wurden 10 Patienten mit T2DM und 15 Gesunden Probanden Muskelgewebe extrahiert. In der anschließenden Zytophotometrie erfolgte die Bestimmung der glykolytischen und oxidativen Enzymaktivität in Abhängigkeit der Fasercharakteristik nach SO, FOG und FT-Fasern. Die Untersuchung verdeutlichte eine Verminderung der oxidativen Enzymaktivität des M. vastus lateralis im Homogenat bei bestehenden T2DM mit gleichzeitiger Verringerung der SO- Muskelfasern um 16 Prozent und Erhöhung der FT- Muskelfasern um 49 Prozent im Vergleich zur Kontrollpopulation. Bei T2DM ist sowohl die oxidative als auch die glykolytische Enzymaktivität in den FG-Fasern als auch in den FOGMischfasern signifikant erhöht. Zusammenfassend weisen unsere Ergebnisse darauf hin, dass die geringere oxidative Enzymaktivität im Homogenat des Skelettmuskel von Patienten mit T2DM vielmehr durch einen verminderten Anteil von SO-Fasern als durch verminderte oxidative Aktivität in einzelnen Fasern verursacht ist. Die erhöhte glykolytische und oxidative Enzymaktivität in einzelnen Muskelfasern korreliert mit dem Maß langfristiger BZHomöostase und der Insulinempfindlichkeit. Diese Adaptation der Skelettmuskulatur könnte einen kompensatorischen Mechanismus bezüglich des pathologischen Glukosestoffwechsels des T2DM darstellen.

The main goal of this study was to assess the impact that nitrate and light have on the relationship between the gene expression of the enzyme nitrate reductase and the incorporation of nitrate in the cosmopolitan diatom Thalassiosira pseudonana, both in laboratory experiments and in natural environments. Continuous cultures were grown at different nitrate (NO3-) concentrations (i.e., 60, 120, and 400 µM) to evaluate their effects on the expression levels of different genes of the nitrogen metabolic pathway (i.e., nitrate and ammonium transporters, nitrate and nitrite reductases, glutamine synthetases II and III). Semi-continuous cultures were grown under different irradiances (i.e., 50, 110, 200, and 320 µmol photon cm-2 s-1) to assess the influence of light intensity (irradiance) on the relationship between the expression of those genes, uptake, and assimilation of nitrate. The expression of all of the genes that were tested decreased significantly (p < 0.05) at the highest concentration of NO3- (i.e., 400 µM), with nitrate transporters showing the most pronounced change from 27.97 to 0.59 fold change cell-1 x 10-6, at 60 and 400 µM NO3- concentrations respectively. Ammonium transporters were detected at all concentrations of NO3-, suggesting that cells are always ready to metabolize ammonium. Growth was limited (µ = 0.99 d-1) by the low irradiance treatment, was maximum (µ = 2.04 d-1) at 200 µmol photon cm-2 s-1 and was inhibited (µ = 1.54 d-1) at the highest irradiance. These trends were reflected in gene expression and uptake rates, with minimum values at the lowest and highest irradiance levels. However, results from the enzymatic assay did not show any significant differences between treatments (p > 0.05). The trends observed in the enzymatic rates could be explained by the gene expression of NO3- reductase and the uptake and growth rates in a multiple regression analysis (R2 = 0.66, p < 0.05). The results of this study show that uptake is independent of gene expression, probably because of a decoupling between transcription and protein synthesis. Not all of the newly synthesized transcripts will inevitably be translated into proteins. And even if they were, there could be post-translational mechanisms preventing the enzymes to become active. This indicates that uptake can be independent of the expression. It was attempted to measure the expression of T. pseudonana genes involved in the metabolism of NO3- in natural diatom assemblages. The use of gene expression as a proxy for metabolic processes carried out by a phytoplankton assemblage in the field is limited and depends on environmental factors, since the current methods of assessing expression rely on genomic sequences that are particularly variable in phytoplankton. The assessment of gene expression provides a useful insight into physiological studies of phytoplankton, and it should be complemented with other measurements, such as the biomass and taxonomic composition of the assemblage for a more complete picture of marine ecosystem nutrient dynamics. / Graduate

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniv., 2007. / Härtill 4 uppsatser.

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniv. / Härtill 4 uppsatser.

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniv., 2006. / Härtill 4 uppsatser och 1 patent.

Magister Scientiae - MSc (Biotechnology) / Glycine max (soybean) is an important crop species globally as it is used as a protein-rich food and feed crop and as a source of oils used in the food and biofuel industry. However, the growth and yield of soybean is adversely affected by drought. Exposure of soybean to drought leads to accumulation of reactive oxygen species (ROS) and cell membrane instability. Sterols are membrane components that regulates membrane fluidity and permeability. Besides being major components of the cell membranes, sterols such as lanosterol appear to play a role in the regulation of ROS scavenging and some are precursors to brassinosteroids that act as signaling molecules with hormonal function that regulate growth, development and responses to abiotic stresses such as drought and salinity. In this study, the involvement of plant sterols, also known as phytosterols, in the regulation of soybean responses to drought stress was investigated in Glycine max by determining the effects of drought on the expression of a candidate lanosterol synthase gene (Glyma08g24160) and the content of a subset of phytosterols in soybean. The effects of inhibition of sterol synthesis on ROS production and on superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT) and dehydroascorbate reductase (DHAR) were investigated. The concentration of hydrogen peroxide (H2O2) as well as superoxide (O2-) increased in response to drought and sterol synthesis inhibition, however, O2- concentration and sterol contents declined under drought stress and sterol synthesis inhibition.