Full-scale biological phosphorus removal: quantification of storage polymers, microbial performance and metabolic modelling

Dissertação para obtenção do Grau de Doutor em Engenharia Química e Bioquímica

Diffusion of the synthetic pyrethroid permethrin into bed-sediments

Bed-sediments are a sink for many micro-organic contaminants in aquatic environments. The impact of toxic contaminants on benthic fauna often depends on their spatial distribution, and the fate of the parent compounds and their metabolites. The distribution of a synthetic pyrethroid, permethrin, a compound known to be toxic to aquatic invertebrates, was studied using river bed-sediments in lotic flume channels. trans/cis-Permethrin diagnostic ratios were used to quantify the photoisomerization of the trans isomer in water. Rates were affected by the presence of sediment particles and colloids when compared to distilled water alone. Two experiments in dark/light conditions with replicate channels were undertaken using natural sediment, previously contaminated with permethrin, to examine the effect of the growth of an algal biofilm at the sediment-water interface on diffusive fluxes of permethrin into the sediment. After 42 days, the bulk water was removed, allowing a fine sectioning of the sediment bed (i.e., every mm down to 5 mm and then 5-10 mm, then every 10 mm down to 50 mm). Permethrin was detected in all cases down to a depth of 5-10 mm, in agreement with estimates by the Millington and Quirk model, and measurements of concentrations in pore water produced a distribution coefficient (K-d) for each section, High K-d's were observed for the top layers, mainly as a result of high organic matter and specific surface area. Concentrations in the algal biofilm measured at the end of the experiment under light conditions, and increases in concentration in the top 1 mm of the sediment, demonstrated that algal/bacterial biofilm material was responsible for high K-d's at the sediment surface, and for the retardation of permethrin diffusion. This specific partition of permethrin to fine sediment particles and algae may enhance its threat to benthic invertebrates. In addition,the analysis of trans/cis-permethrin isomer ratios in sediment showed greater losses of trans-permethrin in the experiment under light conditions, which may have also resulted from enhanced biological activity at the sediment surface.

Enhanced priming for trauma-related material in posttraumatic stress disorder

Intrusive reexperiencing in posttraumatic stress disorder (PTSD) has been linked to perceptual priming for trauma-related material. A prospective longitudinal study (N = 69) investigated perceptual priming for trauma-related, general threat, and neutral words in assault survivors with and without PTSD, using a new version of the word-stem completion task. Survivors with PTSD showed enhanced priming for trauma-related words. Furthermore, priming for trauma-related words measured soon after the trauma was associated with subsequent PTSD severity at 3 6, and 9 months. The enhanced priming effect was specific to trauma-related words. Enhanced perceptual priming for traumatic material appears to be one of the cognitive processes operating in PTSD.

Biological effects of Ocimum gratissimum L. are due to synergic action among multiple compounds present in essential oil

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

The gastroprotective effect of the essential oil of Croton cajucara is different in normal rats than in malnourished rats

It has been shown previously that malnourished rats are resistant to acute gastric lesions but not to subchronic gastric ulceration. It also has been demonstrated that the essential oil obtained from the bark of Croton cajucara (Sacaca) has antiulcer properties. In the present study, the ability of this essential oil to prevent the formation of gastric ulcers in rats fed a diet with 17 % protein (normoproteic rats) or 6 % protein (malnourished rats) was investigated. At a dose of 100 mg/kg body weight, orally, the essential oil significantly reduced the gastric injury caused by indomethacin (25 % after 2 h and 70 % after 4 h) only in normoproteic rats. In the pylorus ligature model, the essential oil increased the pH and gastric volume, but decreased the total acid concentration in both groups when compared to the respective control group. The essential oil significantly increased prostaglandin E2 production in glandular cells by 50 % compared to the controls in both groups of rats. In addition, the amount of gastric mucus was two-fold higher in malnourished rats than in normoproteic rats. The present results show that the enhanced protective effect of essential oil in malnourished rats involved an increase in prostaglandin E2 production and mucus secretion, which are both factors that protect the gastric mucosa against damage. In agreement with this, malnourished rats always had a lower number of acute gastric ulcers.

In vitro Remineralizing Effect of Fluoride Varnishes Containing Sodium Trimetaphosphate

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Synergistic effect of photodynamic therapy and cisplatin: a novel approach for cervical cancer

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Design of cell adhesive and angiogenic titanium surfaces for cellular stimulation

Die Förderung der Zelladhäsion durch sogenannte biomimetische Oberflächen wird in der Medizin als vielversprechender Ansatz gesehen, um Komplikationen wie z. B. Fremdkörperreaktionen nach der Implantation entgegenzuwirken. Neben der Immobilisierung einzelner Biomoleküle wie z. B. dem RGD-Peptid, Proteinen und Wachstumsfaktoren auf verschiedenen Materialien, konzentriert man sich derzeit in der Forschung auf die Co-Immobilisierung zweier Moleküle gleichzeitig. Hierbei werden die funktionellen Gruppen z. B. von Kollagen unter Verwendung von nur einer Kopplungschemie verwendet, wodurch die Kopplungseffizienz der einzelnen Komponenten nur begrenzt kontrollierbar ist. Das Ziel der vorliegenden Arbeit war die Entwicklung eines Immobilisierungsverfahrens, welches die unabhängige Kopplung zweier Faktoren kontrolliert ermöglicht. Dabei sollten exemplarisch das adhäsionsfördernde RGD-Peptid (Arginin-Glycin-Asparaginsäure) zusammen mit dem Wachstumsfaktor VEGF (Vascular Endothelial Growth Factor) auf Titan gebunden werden. In weiteren Experimenten sollten dann die pro-adhäsiven Faktoren Fibronektin, Kollagen, Laminin und Osteopontin immobilisiert und untersucht werden. rnDie Aminofunktionalisierung von Titan durch plasma polymerisierte Allylaminschichten wurde als Grundlage für die Entwicklung des nasschemischen Co-immobilisierungsverfahren verwendet. Für eine unabhängige und getrennte Anbindung der verschiedenen Biomoleküle stand in diesem Zusammenhang die Entwicklung eines geeigneten Crosslinker Systems im Vordergrund. Die Oberflächencharakterisierung der entwickelten Oberflächen erfolgte mittels Infrarot Spektroskopie, Surface Plasmon Resonance Spektroskopie (SPR), Kontaktwinkelmessungen, Step Profiling und X-Ray Photoelectron Spektroskopie (XPS). Zur Analyse der Anbindungsprozesse in Echtzeit wurden SPR-Kinetik Messungen durchgeführt. Die biologische Funktionalität der modifizierten Oberflächen wurde in vitro an Endothelzellen (HUVECs) und Osteoblasten (HOBs) und in vivo in einem Tiermodell-System an der Tibia von Kaninchen untersucht.rnDie Ergebnisse zeigen, dass alle genannten Biomoleküle sowohl einzeln auf Titan kovalent gekoppelt als auch am Bespiel von RGD und VEGF in einem getrennten Zwei-Schritt-Verfahren co-immobilisiert werden können. Des Weiteren wurde die biologische Funktionalität der gebundenen Faktoren nachgewiesen. Im Falle der RGD modifizierten Oberflächen wurde nach 7 Tagen eine geförderte Zelladhäsion von HUVECs mit einer signifikant erhöhten Zellbesiedlungsdichte von 28,5 % (p<0,05) gezeigt, wohingegen auf reinem Titan Werte von nur 13 % beobachtet wurden. Sowohl VEGF als auch RGD/VEGF modifizierte Proben wiesen im Vergleich zu Titan schon nach 24 Stunden eine geförderte Zelladhäsion und eine signifikant erhöhte Zellbesiedlungsdichte auf. Bei einer Besiedlung von 7,4 % auf Titan, zeigten VEGF modifizierte Proben mit 32,3 % (p<0,001) eine deutlichere Wirkung auf HUVECs als RGD/VEGF modifizierte Proben mit 13,2 % (p<0,01). Die pro-adhäsiven Faktoren zeigten eine deutliche Stimulation der Zelladhäsion von HUVECs und HOBs im Vergleich zu reinem Titan. Die deutlich höchsten Besiedlungsdichten von HUVECs konnten auf Fibronektin mit 44,6 % (p<0,001) und Kollagen mit 39,9 % (p<0,001) nach 24 Stunden beobachtet werden. Laminin zeigte keine und Osteopontin nur eine sehr geringe Wirkung auf HUVECs. Bei Osteoblasten konnten signifikant erhöhte Besiedlungsdichten im Falle aller pro-adhäsiven Faktoren beobachtet werden, jedoch wurden die höchsten Werte nach 7 Tagen auf Kollagen mit 90,6 % (p<0,001) und Laminin mit 86,5 % (p<0,001) im Vergleich zu Titan mit 32,3 % beobachtet. Die Auswertung der Tierexperimente ergab, dass die VEGF modifizierten Osteosyntheseplatten, im Vergleich zu den reinen Titankontrollen, eine gesteigerte Knochenneubildung auslösten. Eine solche Wirkung konnte für RGD/VEGF modifizierte Implantate nicht beobachtet werden. rnInsgesamt konnte gezeigt werden, dass mittels plasmapolymerisierten Allylamin Schichten die genannten Biomoleküle sowohl einzeln gebunden als auch getrennt und kontrolliert co-immobilisiert werden können. Des Weiteren konnte eine biologische Funktionalität für alle Faktoren nach erfolgter Kopplung in vitro gezeigt werden. Wider Erwarten konnte jedoch kein zusätzlicher biologischer Effekt durch die Co-immobilisierung von RGD und VEGF im Vergleich zu den einzeln immobilisierten Faktoren gezeigt werden. Um zu einer klinischen Anwendung zu gelangen, ist es nun notwendig, das entwickelte Verfahren in Bezug auf die immobilisierten Mengen der verschiedenen Faktoren hin zu optimieren. rn

The effect of RNA base lesions on mRNA translation

The biological effect of oxidatively damaged RNA, unlike oxidatively damaged DNA, has rarely been investigated, although it poses a threat to any living cell. Here we report on the effect of the commonly known RNA base-lesions 8-oxo-rG, 8-oxo-rA, ε-rC, ε-rA, 5-HO-rC, 5-HO-rU and the RNA abasic site (rAS) on ribosomal translation. To this end we have developed an in vitro translation assay based on the mRNA display methodology. A short synthetic mRNA construct containing the base lesion in a predefined position of the open reading frame was 32P-labeled at the 5′-end and equipped with a puromycin unit at the 3′-end. Upon in vitro translation in rabbit reticulocyte lysates, the encoded peptide chain is transferred to the puromycin unit and the products analyzed by gel electrophoresis. Alternatively, the unlabeled mRNA construct was used and incubated with 35S-methionine to prove peptide elongation of the message. We find that all base-lesions interfere substantially with ribosomal translation. We identified two classes, the first containing modifications at the base coding edge (ε-rC, ε-rA and rAS) which completely abolish peptide synthesis at the site of modification, and the second consisting of 8-oxo-rG, 8-oxo-rA, 5-HO-rC and 5-HO-rU that significantly retard full-length peptide synthesis, leading to some abortive peptides at the site of modification.

Leptin can induce proliferation, differentiation, and functional activation of hemopoietic cells

Many cytokines exert their biological effect through members of the hemopoietin receptor family. Using degenerate oligonucleotides to the common WSXWS motif, we have cloned from human hemopoietic cell cDNA libraries various forms of the receptor that was recently shown to bind the obesity hormone, leptin. mRNAs encoding long and short forms of the human leptin receptor were found to be coexpressed in a range of human and murine hemopoietic organs, and a subset of cells from these tissues bound leptin at the cell surface. Ectopic expression in murine Ba/F3 and M1 cell lines revealed that the long, but not the short, form of the leptin receptor can signal proliferation and differentiation, respectively. In cultures of murine or human marrow cells, human leptin exhibited no capacity to stimulate cell survival or proliferation, but it enhanced cytokine production and phagocytosis of Leishmania parasites by murine peritoneal macrophages. Our data provide evidence that, in addition to its role in fat regulation, leptin may also be able to regulate aspects of hemopoiesis and macrophage function.

Applicability of experience from laboratory reactors with biological phosphorus removal in full-scale plants

Enhanced biological phosphorus removal (EBPR) has been used at many wastewater treatment plants all over the world for many years. In this study a full-scale sludge with good EBPR was tested with P-release batch tests and combined FISH/MAR (fluorescence in situ hybridisation and microautoradiography). Proposed models of PAOs and GAOs (polyphosphate- and glycogen-accumulating organisms) and microbial methods suggested from studies of laboratory reactors were found to be applicable also on sludge from full-scale plants. Dependency of pH and the uptake of both acetate and propionate were studied and used for calculations for verifying the models and results from microbial methods. All rates found from the batch tests with acetate were higher than in the batch tests with propionate, which was explained by the finding that only those parts of the bacterial community that were able to take up acetate anaerobically were able to take up propionate anaerobically.

Isolation and characterisation of conomap-Vt, a D-amino acid containing excitatory peptide from the venom of a vermivorous cone snail

Cone snail venom is a rich source of bioactives, in particular small disulfide rich peptides that disrupt synaptic transmission. Here, we report the discovery of conomap-Vt (Conp-Vt), an unusual linear tetradecapeptide isolated from Conus vitulinus venom. The sequence displays no homology to known conopeptides, but displays significant homology to peptides of the MATP (myoactive tetradecapeptide) family, which are important endogenous neuromodulators in molluscs, annelids and insects. Conp-Vt showed potent excitatory activity in several snail isolated tissue preparations. Similar to ACh, repeated doses of Conp-Vt were tachyphylactic. Since nicotinic and muscarinic antagonists failed to block its effect and Conp-Vt desensitised tissue remained responsive to ACh, it appears that Conp-Vt contractions were non-cholinergic in origin. Finally, biochemical studies revealed that Conp-Vt is the first member of the MATP family with a D-amino acid. Interestingly, the isomerization of L-Phe to D-Phe enhanced biological activity, suggesting that this post-translational modified conopeptide may have evolved for prey capture. (c) 2006 Published by Elsevier B.V. on behalf of the Federation of European Biochemical Societies.

Modelling biological processes under anaerobic conditions through integrating titrimetric and off-gas measurements - applied to EBPR systems

An innovative method for modelling biological processes under anaerobic conditions is presented and discussed. The method is based on titrimetric and off-gas measurements. Titrimetric data is recorded as the addition rate of hydroxyl ions or protons that is required to maintain pH in a bioreactor at a constant level. An off-gas analysis arrangement measures, among other things, the transfer rate of carbon dioxide. The integration of these signals results in a continuous signal which is solely related to the biological reactions. When coupled with a mathematical model of the biological reactions, the signal allows a detailed characterisation of these reactions, which would otherwise be difficult to achieve. Two applications of the method to the enhanced biological phosphorus removal processes are presented and discussed to demonstrate the principle and effectiveness of the method.

Enhanced zinc oxide and graphene nanostructures for electronics and sensing applications

Zinc oxide and graphene nanostructures are important technological materials because of their unique properties and potential applications in future generation of electronic and sensing devices. This dissertation investigates a brief account of the strategies to grow zinc oxide nanostructures (thin film and nanowire) and graphene, and their applications as enhanced field effect transistors, chemical sensors and transparent flexible electrodes. Nanostructured zinc oxide (ZnO) and low-gallium doped zinc oxide (GZO) thin films were synthesized by a magnetron sputtering process. Zinc oxide nanowires (ZNWs) were grown by a chemical vapor deposition method. Field effect transistors (FETs) of ZnO and GZO thin films and ZNWs were fabricated by standard photo and electron beam lithography processes. Electrical characteristics of these devices were investigated by nondestructive surface cleaning, ultraviolet irradiation treatment at high temperature and under vacuum. GZO thin film transistors showed a mobility of ∼5.7 cm2/V·s at low operation voltage of <5 V and a low turn-on voltage of ∼0.5 V with a sub threshold swing of ∼85 mV/decade. Bottom gated FET fabricated from ZNWs exhibit a very high on-to-off ratio (∼106) and mobility (∼28 cm2/V·s). A bottom gated FET showed large hysteresis of ∼5.0 to 8.0 V which was significantly reduced to ∼1.0 V by the surface treatment process. The results demonstrate charge transport in ZnO nanostructures strongly depends on its surface environmental conditions and can be explained by formation of depletion layer at the surface by various surface states. A nitric oxide (NO) gas sensor using single ZNW, functionalized with Cr nanoparticles was developed. The sensor exhibited average sensitivity of ∼46% and a minimum detection limit of ∼1.5 ppm for NO gas. The sensor also is selective towards NO gas as demonstrated by a cross sensitivity test with N2, CO and CO2 gases. Graphene film on copper foil was synthesized by chemical vapor deposition method. A hot press lamination process was developed for transferring graphene film to flexible polymer substrate. The graphene/polymer film exhibited a high quality, flexible transparent conductive structure with unique electrical-mechanical properties; ∼88.80% light transmittance and ∼1.1742Ω/sq k sheet resistance. The application of a graphene/polymer film as a flexible and transparent electrode for field emission displays was demonstrated.