<p dir="ltr">Subcutaneous injection for self-administration of biotherapeutics, such as monoclonal antibodies (mAbs), is becoming increasingly prominent within the pharmaceutical sector due to its benefits in patient compliance and cost-effectiveness. The success of this drug delivery process depends on the coupled mechanical and transport phenomena within the subcutaneous tissue, both during and after the injection. Yet, the details of these processes are not well-elucidated, sparking a surge in computational efforts to fill this knowledge gap. Remarkably, there are very few computational studies on subcutaneous injection into three-dimensional porous media that account for large tissue deformations, drug transport and absorption, the use medical devices, and human factors. Here, we develop a high-fidelity computational framework to study large-volume subcutaneous injection of mAbs. Our investigation begins with a linear poroelastic model without drug transport, which we employ to study the effect of tissue deformation on injection dynamics. We progressively enhance this model, advancing to a nonlinear porohyperelastic framework that include drug transport and absorption. To capture the anisotropy of subcutaneous tissue, we employ a fibril-reinforced porohyperelastic model. Furthermore, we integrate the multi-layered structure of skin tissue by creating data-driven geometrical models of the tissue layers derived from histological data. Our analysis explores the impact of different handheld autoinjectors on the injection dynamics for various patient-applied forces. We investigate the effect of different pre-injection techniques, such as the pinch and stretch methods, on the drug transport and absorption. Additionally, we evaluate the impact of several physiological variables, including flow rate, injection depth, and body mass index. Our simulations yield crucial insights essential for comprehending and improving subcutaneous drug administration of mAbs. Additionally, they offer a deeper understanding of the human aspect of the injection procedure, thereby paving the way for advancements in the development of patient-centered injection devices and techniques.</p>

Environmental contamination issues related to phosphorus (P) in surface waters substantiates the need to identify minimally-sufficient P fertilization amounts for production of containerized nursery crops and better understand the effect of routine amendments (i.e., dolomite [DL] and micronutrient fertilizer [MF]) added to pine bark substrates on chemical fate of P fertilizer. Four studies were conducted to accomplish two overarching objectives: 1) determine the minimum P fertilization amount and corresponding pore-water P concentration needed to achieve maximal growth of common containerized nursery crops and 2) determine the effect of DL and MF amendments in pine bark on P retention during irrigation and P fractions in substrate pore-water. In a fertigation, greenhouse study, calculated lowest P-fertilizer concentration that sustained maximal growth in Hydrangea paniculata ‘Limelight’ (panicle hydrangea) and Rhododendron ‘Karen’ (azalea) was 4.7 and 2.9 mg·L⁻¹ , respectively, and shoot growth Ilex crenata ‘Helleri’ (holly) was the same when fertilized with 0.5 to 6.0 mg·L⁻¹ P. Porewater P concentrations corresponding with treatments that sustained maximal growth of panicle hydrangea, azalea and holly were as low as 0.6, 2.2 and 0.08 mg·L⁻¹ P, respectively. In a separate study, utilizing low-P controlled-release fertilizers (CRFs), shoot growth of Hydrangea macrophylla ‘P11HM-11’ (bigleaf hydrangea) produced in two ecoregions was maximal when fertilized with as little as 0.3 g CRF-P per 3.8-L container, a 50% P reduction from the industrystandard CRF. Holly required 0.2 or 0.4 g CRF-P depending on ecoregion. Mean pore-water P concentrations that corresponded with highest SDW were 0.8 and 1.2 mg·L⁻¹ for hydrangea and holly, respectively. When irrigating fallow pine bark columns containing CRF for 48 d, amending pine bark with DL and MF reduced orthophosphate-P (OP-P) leachate concentrations by ≈ 70%, most of which was retained within the substrate. In a greenhouse study, containerized Lagerstroemia ‘Natchez’ (crape myrtle) were grown for 91 d in pine bark containing CRF. In pine bark amended with DL and MF, pore-water OP-P and total P concentrations, measured approximately weekly, were reduced by, on average, 64% and 58%, respectively. Total dry weight values of plants grown with DL plus MF or MF-only were 40% higher than those grown with no amendments; however, tissue P amounts and relative P uptake efficiency were the same among plants in these three treatments. Therefore, sorption of OP-P by DL and MF reduced water-extractable OP-P but did not limit P uptake by plants. / Ph. D. / Phosphorus (P) pollution of surface waters is a global issue that has led to widespread fish kills, drinking water contamination and disruption of aquatic ecosystems. Nutrient runoff from agricultural sites is among the leading contributors to P loads in impaired waters. Optimizing P fertilization for containerized nursery crop production is particularly challenging since the primary soilless substrates used to grow containerized crops retain P poorly. Consequently, much of the applied P leaches from containers during irrigation. Reducing amounts of applied P fertilizer and amending substrates (e.g., pine bark) with P-sorbing materials are two methods previously shown to reduce P leaching and increase the proportion of applied P that is absorbed by containerized plants. Four studies were conducted to accomplish two overarching objectives: 1) determine the minimum P fertilization amount necessary for maximal growth of common containerized nursery crops and 2) determine the effect of dolomite (DL) and micronutrient fertilizer (MF) amendments in pine bark on P retention during irrigation. Our findings indicated that P fertilization requirements of woody ornamental crops is species-dependent. When using liquid fertilizer, Japanese holly and evergreen azalea achieved maximal growth when P fertilizer concentrations were reduced by 90% and 40%, respectively, compared to current recommendations. In contrast, the current minimum fertilizer recommendation of 5 ppm P was optimal for panicle hydrangea. In a subsequent study in which containerized woody ornamentals were grown using low-P controlled-release fertilizers (CRFs), bigleaf hydrangea reached maximal growth when given CRFs containing 50% less P than amounts in conventional CRFs. Considering hydrangea and azalea are among the top woody ornamental shrubs produced in the US, using fertilizers with minimally sufficient P amounts for these species could greatly reduce P runoff from nursery sites. Results of two studies conducted to achieve the second aforementioned objective indicated that amending CRF-fertilized pine bark with DL and MF can reduce water-extractable total P concentrations by > 50%. Despite lower levels of plant-available P in the substrate, P uptake by crape myrtle was unaffected by the amendments. The DL was primarily responsible for P retention in pine bark; however, the addition of MF was needed for maximal growth and P uptake of crape myrtle. According to this research, amending pine bark with DL and MF could be considered a best management practice for reducing P leaching from containerized crops.

No / Mono- and disaccharide-functionalised conjugates of the fluoroquinolone antibiotic ciprofloxacin have been synthesised and used as chemical probes of the bacterial uptake of glycosylated ciprofloxacin. Their antimicrobial activities against a panel of clinically relevant bacteria were determined: the ability of these conjugates to inhibit their target DNA gyrase and to be transported into the bacteria was assessed by using in vivo and in vitro assays. The data suggest a lack of active uptake through sugar transporters and that although the addition of monosaccharides is compatible with the inhibition of DNA gyrase, the addition of a disaccharide results in a significant decrease in antimicrobial activity.

In der vorliegenden Arbeit wurde die Regulation der Aktivität der vesikulären Monoamintransporter VMAT1 und VMAT2 durch heterotrimere G-Proteine untersucht. In der humanen neuroendokrinen Zellinie BON werden VMAT1 und VMAT2 exprimiert. Sie colokalisieren in diesen Zellen mit der a-Untereinheit des heterotrimeren G-Proteins Go2 vorwiegend auf großen elektronendichten Vesikeln, den LDCVs. Die Aktivität beider Transporter unterliegt einer Regulation durch Gao2. Nach Aktivierung des G-Proteins kommt es zu einer Hemmung der vesikulären Monoaminaufnahme. Die Aktivität von VMAT2 wird dabei empfindlicher reguliert als die Aktivität von VMAT1. In Primärkulturen von Rapheneuronen der Ratte wird VMAT2 als neuronale Variante des Transporters exprimiert. VMAT2 lokalisiert in diesen Neuronen überwiegend auf kleinen synaptischen Vesikeln, den SSVs. Hier kommt es zu einer Colokalisation mit Gao2 auf diesem Vesikeltyp. Auch in Rapheneuronen wird die Aktivität von VMAT2 durch diese G-Protein Untereinheit gehemmt. Elektronenmikroskopische Befunde belegen die Lokalisation von VMAT2 und Gao2 auf SSVs von serotonergen Axonterminalen im präfrontalen Cortex der Ratte. An einer Präparation synaptischer Vesikel aus diesem Gehirnbereich konnte ebenfalls eine Hemmung der Transportaktivität von VMAT2 durch Gao2 nachgewiesen werden. Auch in Thrombozyten der Maus unterliegt die vesikuläre Serotoninaufnahme einer Hemmung durch ein heterotrimeres G-Protein. In chronisch entleerten Vesikeln aus Mäusen, in denen das Gen für die periphere Tryptophanhydroxylase deletionsmutiert vorlag, konnte zunächst keine Hemmung der Serotoninaufnahme durch heterotrimere G-Proteine beobachtet werden. Nach Vorbeladung der Vesikel mit Serotonin war dies jedoch der Fall. Die Aktivierung des G-Proteins wird somit sehr wahrscheinlich über den Füllungszustand der Vesikel gesteuert. / In this study we investigated the regulation of the activity of the vesicular monoamine transporters VMAT1 and VMAT2 by heterotrimeric G-proteins. In the human neuroendocrine cell line BON both transporters are expressed. They colocalize in these cells with the a-subunit of the heterotrimeric G-protein Go2 predominantely on Large Dense Core Vesicles (LDCVs). The activity of both VMAT1 and VMAT2 is regulated by Gao2. G-protein activation results in a down-regulation of vesicular monoamine uptake. VMAT2 appears to be more sensitive towards the observed G-protein regulation than VMAT1. Serotonergic raphe neurons in primary culture express VMAT2 as the neuronal form of the transporter. In these neurons VMAT2 predominantely localizes to Small Synaptic Vesicles (SSVs). Here, VMAT2 colocalizes with Gao2 on SSVs. In these neurons Gao2-dependent down-regulation of VMAT2 activity was observed, too. Immunoelectron microscopic analysis confirmed a localization of VMAT2 and Gao2 on SSVs from serotonergic terminals in the rat prefrontal cortex. In addition, Gao2-dependent regulation of VMAT2 activity could also be demonstrated when using a crude synaptic vesicle preparation of this brain area. Even in platelets obtained from mice the vesicular serotonin uptake is down-regulated by heterotrimeric G-proteins. In serotonin-depleted platelets from peripheral tryptophane-hydroxylase knockout mice no G-protein-dependent down-regulation of monoamine uptake was observed. After preincubation of the platelets with serotonin, the G-protein regulation was restored. Therefore, the vesicular transmitter content appears to be a likely factor of G-protein activation in platelets.

Sterine sind essentielle Komponenten der Zellmembran, deren Konzentration und Lokalisierung genau kontrolliert wird. Die Hefe Saccharomyces cerevisiae ist ein fakultativ anaerober Organismus, der in Abwesenheit von Sauerstoff auxotroph für Sterine wird. Die Proteine Aus1p und Pdr11p gehören zur Familie der ABC Proteine und spielen eine wichtige Rolle in diesem Prozess, da die gleichzeitige Deletion beider Protein die Aufnahme von Sterinen unter anaeroben Wachstumsbedingungen blockiert.In dieser Arbeit wurde das Gen AUS1 in voller Länge kloniert. Methoden für die Extraktion und Reinigung dieses Transporters wurden entwickelt, damit dieser detailliert charakterisiert werden kann. Mit Hilfe von Detergenzien wurde das Protein löslich gemacht und zeigte ATP-Bindung und -Hydrolyse. Die ATP-Hydrolyse konnte durch die Mutation eines konservierten Lysins zu Methionin im Walker A Motif verhindert. Genauso konnte die ATP-Hydrolyse auch durch klassische Inhibitoren von ABC Transportern inhibiert werden. Nach der Rekonstitution von Aus1p in Proteoliposomen wurde die ATPase Aktivität spezifisch durch Phosphatidylserin in einer stereoselektiven Weise stimuliert.Zusätzlich konnte gezeigt werden, dass Änderungen im zellulären PS Spiegel die Aus1p-abhängige Aufnahme von Sterin an die Membran beeinflussen. Diese Ergebnisse schlagen eine für die Aktivität des Transporters wichtige, direkte Interaktion zwischen Aus1p und PS vor.Da es sich bei der Aufnahme von Sterin um einen komplexen Prozess handelt, könnten Komponenten exisitieren, die mit Aus1p interagieren. Der Hefestamm, der die Immunpräzipitation von Aus1p mit seinem Interaktionspartner ermöglicht, wurde erzeugt und der Einfluß von Mannoproteinen auf Sterinaufnahme wurde getestet. Außerdem wurde eine Methode entwickelt, mit der Aus1p in Giant Unilamellar Vesicles rekonstituiert werden kann. Mit diesen Liposomen kann das Verhalten und die Aktivität von Aus1p in Membranen mit einer komplexen Lipidzusammensetzung untersucht werden. / Sterols are essential components of cellular membranes and their concentration and localization are tightly controlled. Saccharomyces cerevisiae is a facultative anaerobic organism which becomes auxotrophic for sterols in the absence of oxygen. However, the precise mechanism of sterol uptake remains to be revealed. Two proteins belonging to ABC protein family, Aus1p and Pdr11p were proposed to play a critical role in this process as simultaneous deletion of both of them blocks sterol uptake under anaerobiosis. In the present work, the full length AUS1 gene was cloned. An extraction and purification procedures were then developed to allow for detailed characterization of the transporter. The detergent solubilized protein was shown to bind and hydrolyse ATP. Mutagenesis of the conserved lysine to methionine in the Walker A motif abolished ATP hydrolysis. Likewise, ATP hydrolysis was inhibited by classical inhibitors of ABC transporters. Upon reconstitution into proteoliposomes, the ATPase activity of Aus1p was specifically stimulated by phosphatidylserine (PS) in a stereoselective manner. Furthermore, it was demonstrated that Aus1p-dependent sterol uptake, but not Aus1p expression and trafficking to the plasma membrane, was affected by changes in cellular PS levels. These results suggest a direct interaction between Aus1p and PS which is critical for the activity of the transporter. Because of the complexity of sterol incorporation process efforts were made to identify additional components of the sterol uptake machinery that interact with Aus1p protein. The yeast strain allowing for immunopercipitation of Aus1p with its interaction partners was generated and previously proposed influence of mannoproteins on the sterol uptake was tested. Additionally, method was developed to reconstitute Aus1p protein into Giant Unilamellar Vesicles. These liposomes can be used further for testing of the behaviour and activity of Aus1p in the membranes with complex lipid composition.

No description available.

South Africans carry high debt levels and many deal with the threat of over-indebtedness. In particular, the debt situation of the youth is of utmost concern. This study was undertaken to gain an understanding of how the liability usage of households fluctuates over the life course. The main objective was to identify and describe how debt is accumulated by young South Africans and how household characteristics and events may be related to the uptake of household liabilities. This study conducted a comprehensive literature review culminating with the development of a heuristic model that identified variables that may affect household debt uptake. Quantitative statistical analysis techniques were employed on secondary data acquired from the South African Audience Research Foundation’s All Media and Products Survey for the years 1999 until 2013. The findings identified that household debt follows a familiar life cycle pattern. A number of independent variables were shown to affect household debt uptake. Furthermore, certain of these variables are related to the trajectories of the life course. / Taxation / M. Acc. Sci.

Partial Nitritation/Anammox process (deammonification process), by which occurs oxidation of ammonium to nitrogen gas by autotrophic bacteria in anaerobic conditions, considered to be cost-effective and environmentally friendly method of nitrogen removal. Present research work focuses on achieving a high nitrogen removal degree, thanks to Anammox bacteria, while providing the best performance of the ongoing process. Integrated fixed-film activated sludge (IFAS) reactor was supplied with the main stream of the wastewater after UASB reactor, characterized by low concentration of nitrogen and organic matter. The bacteria ability to accommodate, were tested in the biofilm and in the activated sludge, depending on the different stages in which the process were being conducted. Batch test, such as Specific Anammox Activity (SAA), Nitrate Uptake Rate (NUR) and Oxygen Uptake Rate (OUR), were used for the evaluation of activity of various groups of bacteria. On the basis of laboratory analysis verified the values obtained from the batch tests. It was determined that a high degree of nitrogen removal (92% of NH4-N) was achieved thanks to the dominant activity of the Anammox bacteria, with low participation of other groups of bacteria. It was also proved, that Anammox bacteria activity were overwhelming in the biofilm. Dominant role of Ammonium Oxidizing Bacteria (AOB) was associated with high activity of Anammox bacteria, which together satisfyingly out-competed Nitrite Oxidizing Bacteria (NOB) and heterotrophic bacteria. It has been shown that Anammox bacteria quickly adapt to the new conditions and they are able to assume a dominant role, even in the case of inoculation of the reactor with the sludge from SBR. This allows conclude, that in the case of operational problems, the reactor can be supplied from another source, in order not to inhibit the process.

Das Ziel der vorliegenden Arbeit war es, zu untersuchen, ob sich mit der Düngung von Stroh, Gehölzhäcksel, frischem Pferdemist und Grüngutkompost die Ertragsleistung und Nährstoffaufnahme von Leguminosen sowie der Folgefrucht Winterweizen steigern lässt. Darüber hinaus sollte die Aufnahme von düngebürtigem Kohlenstoff durch Rotklee mittels 13-C-angereichertem Stroh quantifiziert werden. Die Verfügbarkeit von Kalium, teilweise auch von Schwefel und Molybdän, wurde vor allem nach der Düngung von Grüngutkompost und von Pferdemist für die untersuchten Leguminosen verbessert, während nach der Düngung von Gehölzhäcksel und Stroh keine signifikanten Effekte auf die Nährstoffaufnahme der Pflanzen festgestellt werden konnten. Die scheinbare Ausnutzung des gedüngten Stickstoffs durch die Leguminosen lag in Abhängigkeit von der Düngung zwischen 0 und 9 % (Gehölzhäcksel < Grüngutkompost < Pferdemist < Stroh). Nur nach der Düngung von Pferdemist zu Saatplatterbse und Ackerbohne verringerte sich der Anteil an symbiotisch fixiertem Luftstickstoff am Spross-N gegenüber der Kontrolle, nicht jedoch die symbiotisch fixierte N-Menge. Der Vorrat an mineralischem Stickstoff im Boden in einer Tiefe von 0 bis 30 cm war unter Ackerbohne und Rotklee ca. 40 Tage nach der Düngung und Einarbeitung von Gehölzhäcksel gegenüber den Vergleichsvarianten und der Kontrolle deutlich vermindert. Ein Einfluss der durch die Düngung bedingten temporären N-Festlegung im Boden auf den Anteil an fixiertem Luftstickstoff am Spross-N konnte bei keiner der untersuchten Leguminosen festgestellt werden. Der nachfolgende Winterweizen reagierte in einem Jahr mit signifikant höherem Kornertrag auf die Düngung von Grüngutkompost zu Ackerbohne und erhöhtem Rohproteingehalt im Korn auf die Düngung von Pferdemist zu Saatplatterbse. Mit Hilfe von 13-C-markiertem Stroh wurde im Freiland eine Assimilation von 0,5 % der mit dem Stroh gedüngten Kohlenstoffmenge durch Rotklee ermittelt. / The aim of this study was to examine, whether the application of wheat straw, pruning, horse manure and green compost improves the yield formation and nutrient uptake of legumes and the succeeding crop winter wheat. Furthermore it should be quantified the amount of carbon in red clover derived from the organic fertilizer by carbon dioxid assimilation. An improved uptake of potassium, partially of sulphur and molybdenum for the legumes could be detected after the application of green compost and horse manure. No effects were found on plant nutrient uptake after the application of straw and pruning. The apparent utilization of the applied nitrogen by organic fertilizer was 0 to 9 %, depending on the fertilization (pruning < green compost < horse manure < straw). The proportion of symbiontically fixed nitrogen decrease in faba bean and in chick-pea after manuring with horse manure compared with the control. However the total amount of symbiotically fixed nitrogen did not decreased. The amount of mineral nitrogen in the soil (depth: 0 to 0,3 m) was significant reduced under faba bean and red clover, 40 days after application and incorporation of pruning. No effect on the symbiotic nitrogen fixation of the legumes was found because of the organic fertilizer induced temporary immobilization of nitrogen in this soil layer. An increased grain yield was determined in the succeeding winter wheat after fertilizing green compost to faba bean and a higher content of crude protein in grain of the wheat was determined after fertilizing horse manure to chick-pea. With the help of a 13C-tracer method it could be estimated, that about 0,5 % of the added carbon with straw was assimilated by red clover under field conditions.

The study aimed to assess factors affecting the uptake of community-based Tuberculosis care (CTBC) as experienced by patients in Lobatse in order to make recommendations to enhance the uptake of CBTC in this area. A cross-sectional study was conducted, using structured interviews amongst 101 TB patients in Lobatse who registered for directly observed treatment (DOT) for TB in the GOB’s health facilities from January 2011 to August 2013. The SPSS (version 21) was used to analyse the data. Univariate logistic regression models were used. Participation in CTBC was an outcome. The main predictors for participation in CBTC included, knowledge and attitudes towards CTBC (p=0.0003), perceived barriers and enablers towards this programme (p=0.0279), and patient satisfaction with this programme (p=0.0315). The research findings pertain to TB services in Lobatse, because the study was conducted in government health facilities implementing the Botswana National Tuberculosis Programme (BNTP) CTBC guidelines only in Lobatse / Health Studies / M.A. (Public Health)