Wetlands within South Africa are an important source of water and nutrients necessary for biological productivity and often the survival of the local people. In a country where the rural communities depend on wetlands for their day to day provision of water, food and materials it has become necessary to understand the functions within wetland systems, so that proper conservation measures can be applied in order to protect and ensure the sustainable use of wetlands. Due to the fact that South Africa has a semi arid climate, thereby affecting the availability of water it is sensible that studies are under-taken in which, more is explored about the water resources, the protection as well as the sustainable use of the wetlands within the region. Despite the fact that the total area which wetlands cover in South Africa is relatively small, the functions which they provide is of fundamental magnitude not only to wildlife but also as an essential part of the human life support system. Wetlands have the ability to regulate regional flow regimes and are often situated in areas of impeded drainage, which may contribute to the regulation of water. It is thus plausible that if headwater/palustrine wetlands are destroyed, many of the streams and rivers which under normal circumstance are perennial, would not only become non- perennial but the consequence of a drought would be far more severe, as well as increasing the risk in flood damage further down the river. In Giants Castle Game Reserve, situated in the KwaZulu-Natal Drakensberg, several palustrine wetlands where studied to identify the geomorphic attributes which contribute to the origin and maintenance of these wetlands. An added motive for the study was the fact that, according to Ramsar, one of the reasons for conserving the Drakensberg wetlands is to ensure the maintenance and production of water quality to KwaZulu-Natal. Seven wetlands within the Boesmans river upper catchment were identified and studied. Soil investigations were undertaken in an effort to determine the driving forces behind the origin and maintenance, as well as to improve the understanding relating to the functioning of the wetlands. The geomorphic attributes which were identified as being important to the genesis and maintenance of wetlands were found to be the following: low relief, soil piping within wetlands, sediment trapping ability of wetlands, the surface roughness of wetlands, channeling within wetlands, organic matter accumulation as well as geological barriers within wetland system. The adaptability of Longmore’s (2001) Hydro-Geomorphic classification to different catchment areas was also tested and was found suitable for these wetlands, although the influence of piping on wetlands evidently requires further investigation and incorporation into classifications. / Dissertation (MA)--University of Pretoria, 2006. / Geography, Geoinformatics and Meteorology / MA / Unrestricted

<p>Mass spectrometry has become the premium chemical identification method. The next advancement for mass spectrometry is the widespread use of mass spectrometers for on-site chemical/biological identification. Ion trap mass spectrometers have emerged as powerful on-site analytical platforms, in spite of limited mass resolution, due to their compatibility with ambient ionization methods and ready implementation of tandem mass spectrometry (MS/MS). However, conventionally operated ion traps are inefficient in accessing the entire tandem mass spectrometry dataspace. By operating the ion trap at a constant trapping voltage, more efficient tandem mass spectrometry scan modes are accessible. The most efficient is to acquire the entire tandem mass spectrometry data space and this work demonstrates three different methods of acquiring this data domain. These methods acquire the data in under a second and the best performing method was implemented in a miniature mass spectrometer without performance decrease. The impact of this device is most powerful when analysis requires the entire ionized sample be considered to determine the identity of the sample. This was shown to be useful for monitoring the lipid metabolism in a model microorganism. </p>

Hemostasis, thrombosis, and inflammation are tightly interconnected processes which may give rise to thrombo-inflammation, involved in infectious and non-infectious acute and chronic diseases, including cardiovascular diseases (CVD). Traditionally, due to its hemostatic role, blood coagulation is isolated from the inflammation, and its critical contribution in the progressing CVD is underrated, until the full occlusion of a critical vessel occurs. Underlying vascular injury exposes extracellular matrix to deposit platelets and inflammatory cells. Platelets being key effector cells, bridge all the three key processes (hemostasis, thrombosis, and inflammation) associated with thrombo-inflammation. Under physiological conditions, platelets remain in an inert state despite the proximity to the endothelium and other cells which are decorated with glycosaminoglycan (GAG)-rich glycocalyx (GAGs). A pathological insult to the endothelium results in an imbalanced blood coagulation system hallmarked by increased thrombin generation due to losses of anticoagulant and cytoprotective mechanisms, i.e., the endothelial GAGs enhancing antithrombin, tissue factor pathwayinhibitor (TFPI) and thrombomodulin-protein C system. Moreover, the loss of GAGs promotes the release of mediators, such as von Willebrand factor (VWF), platelet factor 4 (PF4), and P-selectin, both locally on vascular surfaces and to circulation, further enhancing the adhesion of platelets to the affected sites. Platelet-neutrophil interaction and formation of neutrophil extracellular traps foster thrombo-inflammatory mechanisms exacerbating the cardiovascular disease course. Therefore, therapies which not only target the clotting mechanisms but simultaneously or independently convey potent cytoprotective effects hemming the inflammatory mechanisms are expected to provide clinical benefits. In this regard, we review the cytoprotective protease activated protein C (aPC) and its strong anti-inflammatory effects thereby preventing the ensuing thrombotic complications in CVD. Furthermore, restoring GAGlike vasculo-protection, such as providing heparin-proteoglycan mimetics to improve regulation of platelet and coagulation activity and to suppress of endothelial perturbance and leukocyte-derived pro-inflammatory cytokines, may provide a path to alleviate thrombo-inflammatory disorders in the future. The vascular tissue-modeled heparin proteoglycan mimic, antiplatelet and anticoagulant compound (APAC), dual antiplatelet and anticoagulant, is an injury-targeting and locally acting arterial antithrombotic which downplays collagen- and thrombin-induced and complement-induced activation and protects from organ injury.

It is important for the ecological management of wildlife populations in the Czech Republic to know the composition of species in individual habitats. This will allow predictions on the reactions of wildlife based on evolving changes that will occur sooner or later in these habitats due to human activity and natural change of our environment. Camera traps are a powerful tool of the 21st century with which we are able to observe wildlife without major interventions in their lives. With the right experimental design, camera traps allow us to find out details about wildlife life such as their distribution and habitat preferences, population structure, and their behavior. In this work, I used camera traps to determine the species composition of two groups; ungulates and carnivores in different habitats. This thesis analyzes the habitat preferences and hepls to find out to what extent the spatial patterns of occurrence on these two groups are in the monitored habitats over the course of a couple years. Data collection took place from June 2015 to May 2017 in the Central Bohemian Region northeast of Prague. Despite the fact that the area is densely populated and agriculturally exploited, I was able to capture four species of ungulates and nine species of carnivores on a total of 73 camera traps in this...

The ability to rapidly detect and identify hazardous analytes in the field has become increasingly important. One of the most important analytical detection methods in the field is gas chromatography-mass spectrometry (GC-MS). In this work, a hand-portable GC-MS system is described that contains a miniature toroidal ion trap mass analyzer and a low thermal mass GC. The system is self-contained within the dimensions of 47 x 36 x 18 cm and weighs less than 13 kg. Because the instrument has a small footprint, it was used as the detector for an automated near-real-time permeation testing system. In permeation testing, materials that are used to make individual protective equipment such as gloves, masks, boots, and suits are exposed to hazardous analytes to determine how long the equipment can be worn safely. The system described herein could test five samples simultaneously. A multi-position valve rotated among the various sample streams and delivered time aliquots into the MS for quantitation. Current field air sampling techniques suffer from long desorption times, high pressure drops, artifact formation and water retention. These disadvantages can be avoided by concentrating the analytes in short open tubular traps containing thick films. There are several advantages to using polymer coated capillaries as traps, including fast desorption, inertness and low flow restriction. An air sampling trap was constructed utilizing open tubular traps for the concentration of semi-volatile organic compounds. The system consisted of multiple capillary traps bundled together, providing high sample flow rates. The analytes were desorbed from the multi-capillary bundle and refocused in a secondary trap. The simultaneous focusing and separation effect of a trap subjected to a negative temperature gradient was also explored. In this configuration, analytes were focused because the front of the peak was at a lower temperature than the rear of the peak and, hence, moved slower. In addition to the focusing effect, analytes with different volatilities focused at different temperatures within the gradient, allowing for separation.

Malaise traps are a common tool for collecting insects used by many researchers. Although there have been variations in the models and materials used for Malaise traps, the potential for sampling bias due to mesh gague has been explored inadequately. This study compared coarse and fine mesh Townes model Malaise traps in two habitats on the Grand Staircase-Escalante National Monument. The two habitats next to the Lick Wash trailhead were defined by dominant vegetation type – sagebrush and grasses or Piñon-Juniper. We collected from three sites per habitat type, over three consecutive days in June in both 2006 and 2007. A pair of Malaise traps consisting of one coarse mesh and one fine mesh trap were used at each site in order to compare differences in the diversity and in the average size of individuals collected by each type of Malaise trap. We measured diversity using both presence-absence data such as richness scores and Jaccard's Index of Similarity, and abundance-based measures of comparison, including Simpson's Index of Diversity and non-metric multidimensional scaling. We identified all individuals according to Order, and because of our interest in flies and their abundance, we further identified the Dipter ta to the Family level. Average insect size was determined by categorizing individuals according to one of 14 distinct size-classes. In sum, 71 samples totaling approximately 62,500 insects were identified and sized. Because we sampled from two adjacent habitats, we also discuss beta diversity across the sample sites. Although mesh-size appears to have a significant effect on the diversity of the catch according to some tests, not all of our analysis agrees. In addition, the gain in the amount of diversity collected by incorporating both mesh-sizes may not be worth the costs of that kind of sampling. Other means of collection may adequately make up that difference. Habitat on the other hand was a clear marker for difference in diversity. Size was not found to be significant overall, but there still may be reasons to examine the effect of mesh-size with respect to the Hymenoptera.

This study examined two different aspects of black bear (Ursus americanus) ecology in Utah. First, we determined the post-denning behaviors of female black bears in order to help management agencies protect bears from human disturbances as well as set spring hunts that minimize the taking of females with dependent young. We looked at the timing of den emergence (X = 25 March), the number of days at the den site post emergence (X = 11 days), and departure (X = 8 April) for female black bears in Utah from 2011—2013. We also analyzed the effects of cohort (lone female, female with cubs, and female with yearlings), region of Utah, year, elevation, and weather on emergence, departure, and total number of days at den. Lastly, we describe behaviors observed at the den site. We found that first emergence was significantly correlated with cohort and spring temperature. Departure date was significantly correlated with geographic region, spring temperature during emergence and departure, and temperature the spring and summer before denning. Total number of days at den was significantly correlated with cohort and last frost date from the year before. Bears spent little of the post-denning period outside of the dens (X = 9.8% of total observation time). When outside of dens, bears were often observed walking, lying down, sitting and standing. We also observed unique behaviors, including gathering nest materials, nursing, and ingesting. Dens were frequently visited by other wildlife as well. Second, we analyzed conflict between humans and black bears in Utah. The Utah Division of Wildlife Resources initiated a black bear sightings and encounters database in 2003. We upgraded this database by gathering available records and organizing them into a new database for analysis using Microsoft Access®. From 2003—2013 there were 943 records, with 499 bear-human encounters, 33 incidents, 10 attacks, 208 property damages, 187 sightings, and 6 vehicle collisions. Utah county had the highest number of events (n = 115). The majority of events took place at campsites (n = 363). Summer was the most common season for events (n = 715). Time of day was frequently not reported, but when it was, most events occurred at night (n = 173). We found no significant increase in the number of events over the last ten years. We also found no significant relationship between the number of events per year and drought data. The highest number of events involved single bears (n = 843), and over half of events had food or garbage available for the bear (n = 475).

Diabetes mellitus is a metabolic disease largely due to lifestyle and nutritional imbalance, resulting in insulin resistance, hyperglycemia and vascular complications. Diabetic kidney disease (DKD) is a major cause of end-stage renal failure contributing to morbidity and mortality worldwide. Therapeutic options to prevent or reverse DKD progression are limited. Endothelial and glomerular filtration barrier (GFB) dysfunction and sterile inflammation are associated with DKD. Neutrophil extracellular traps (NETs), originally identified as an innate immune mechanism to combat infection, have been implicated in sterile inflammatory responses in non-communicable diseases. However, the contribution of NETs in DKD remains unknown. Here, we show that biomarkers of NETs are increased in diabetic mice and diabetic patients and that these changes correlate with DKD severity. Mechanistically, NETs promote NLRP3 inflammasome activation and glomerular endothelial dysfunction under high glucose stress in vitro and in vivo. Inhibition of NETs (PAD4 inhibitor) ameliorate endothelial dysfunction and renal injury in DKD. Taken together, NET-induced sterile inflammation promotes diabetes-associated endothelial dysfunction, identifying a new pathomechanism contributing to DKD. Inhibition of NETs may be a promising therapeutic strategy in DKD.

No description available.

Sepsis is a clinical syndrome characterized by the systemic activation of inflammatory and coagulation pathways in response to microbial infection of normally sterile parts of the body. Despite considerable advances in our understanding of sepsis pathophysiology, sepsis remains the leading cause of death in non-coronary intensive care units (ICU) with a global disease burden between 15 and 19 million cases per year (Dellinger et al., 2008). Severe sepsis, defined as sepsis associated with organ dysfunction is associated with mortality rates of 33% to 45%. The incidence of severe sepsis continues to increase by 1.5% per annum due to the aging population, a rise in the prevalence of comorbidities, and the wider use of immunosuppressive agents and invasive procedures (Angus et al., 2001). Over the past several decades, many potential treatments for sepsis have shown early promise, yet have failed to improve survival in over 100 Phase II and Phase III clinical trials (Marshall, 2014) suggesting that some fundamental knowledge is lacking in our understanding of sepsis pathophysiology. Emerging studies on cell-free DNA (cfDNA), DNA released extracellularly into the circulation, demonstrate that cfDNA is a crucial link between inflammation and coagulation . In various conditions characterized by excessive inflammatory responses or aberrant prothrombotic responses, cfDNA has been implicated in exacerbating disease pathology (Atamaniuk, Kopecky, Skoupy, Säemann, & Weichhart, 2012; Fuchs, Brill, & Wagner, 2012; Swystun, Mukherjee, & Liaw, 2011). In clinical sepsis, levels of cfDNA upon admission into the ICU have strong prognostic value in predicting mortality (Dwivedi et al., 2012; Saukkonen et al., 2008). However, it is unclear whether these increases in cfDNA are an epiphenomenon during sepsis progression, or whether cfDNA actively plays a role in sepsis pathophysiology. In this work, in vivo studies were conducted to characterize the role of cfDNA in sepsis, the effects of DNase administration, and the potential mechanism by which cfDNA is released during experimental sepsis. In addition, mortality studies were conducted to identify surrogate markers of death to promote the design of humane and ethical animal studies in conducting sepsis research. Polymicrobial sepsis was induced via a surgical procedure whereby the cecum is exteriorized, ligated and punctured twice to introduce a continuous source of microorganisms, a model termed cecal ligation and puncture (CLP). In our CLP sepsis model, levels of cfDNA increased in a time-dependent manner. These increases accompanied an early pro-inflammatory response marked by increased pro-inflammatory IL-6, a transient increase in anti-inflammatory IL-10, and elevated lung myeloperoxidase (MPO) activity. Septic mice with elevated cfDNA levels also had high bacterial loads in the lungs, blood, and peritoneal cavity fluid. Organ damage was also observed in mice following CLP surgery versus mice subjected to the non-septic sham control surgery marked by increased levels of creatinine and alanine aminotransferase (ALT) indicative of kidney and liver injury, respectively. Histological analyses further confirmed lung and kidney damage following CLP surgery. Changes in coagulation were also observed in septic mice as mice subjected to CLP had sustained increases in thrombin-antithrombin (TAT) complexes. In addition, plasma from CLP-operated mice had increased thrombin generation (i.e. increased endogenous thromin potential, increased peak thrombin, decreased time to peak, and decreased lag time) mediated by FXIIa and enhanced by platelets. Following CLP-induced sepsis, elevations in cfDNA levels accompanied pro-inflammatory and pro-coagulant responses. The effects of in vivo DNase treatment in septic mice were time-dependent. Early DNase treatment when cfDNA levels were low resulted in an exaggerated pro-inflammatory response marked by increased plasma IL-6 levels and increased lung damage. In contrast, delayed DNase treatment at time-points when cfDNA levels were elevated suppressed inflammation characterized by an increase in anti-inflammatory IL-10 and reductions in cfDNA, IL-6, lung MPO, and ALT activity. Furthermore, delayed DNase administration resulted in decreased bacterial load in the lungs, blood, and peritoneal cavity fluid. Delayed DNase treatment also resulted in blunted pro-coagulant responses as levels of TAT complexes were suppressed and thrombin generation from septic mouse plasma was normalized. Moreover, DNase treatment when cfDNA levels were elevated increased survival in CLP-operated mice by 80% and reduced lung and liver damage. These findings suggest that administration of DNase when cfDNA levels are elevated may reduce pro-inflammatory and pro-coagulant responses and that delayed DNase treatment may infer protection in the CLP model of sepsis. One mechanism by which cfDNA is released is via the formation of neutrophil extracellular traps (NETs). Upon inflammatory stimulation, some neutrophils release chromatin material and antimicrobial proteins (i.e. neutrophil elastase, MPO, and histones) in an active process termed NETosis. Although NETs ensnare bacteria and exert antimicrobial properties, NETs may also exert harmful effects on the host by activating inflammation and coagulation. While some in vitro evidence suggest that neutrophils are the main source of cfDNA released following inflammatory stimulation, others have reported that neutrophils are not the main source of circulating cfDNA following septic challenge. To determine whether NETs contribute to cfDNA released during CLP sepsis, genetically modified mice that are incapable of forming NETs, PAD4-/- mice, were used. Levels of cfDNA in PAD-/- mice were significantly lower than cfDNA levels in C57Bl/6 mice following CLP surgery, suggesting that NETs were a source of cfDNA in our model. Levels of IL-6, MPO, and bacterial load in the lungs, blood, and peritoneal cavity were significantly reduced, indicating that NETs exert pro-inflammatory effects in CLP sepsis. Thrombin generation was also suppressed in PAD4-/- mice which suggests that NETs contribute to thrombin generation following CLP sepsis. NETs contribute to increases in circulating cfDNA and may exacerbate pathology by driving pro-inflammatory and pro-coagulant responses in CLP-induced sepsis. Appreciating the implications of conducting research using animals, it is pertinent that researchers ensure the highest ethical standards and design animal studies in the most humane, yet scientifically rigorous manner. Using mortality studies, we validated the utility of physiological and phenotypic markers to assess disease severity and predict death in murine sepsis. Temperature via a rectal probe monitor and sepsis scoring systems which assess components such as orbital tightening, level of consciousness, and activity were effective surrogate markers of death. These tools offer a non-invasive assessment of disease progression which do not artificially exacerbate sepsis pathology and immediate information regarding any changes in the health status. Surrogate markers of death also provide reliable monitoring to meet increasing standards of ethical, humane animal research and a feasible and cost-efficient means to obtain vital signs in small rodents. We have proposed a scoring system which can be used for assessing disease severity, endpoint monitoring, and predicting death to obviate inhumane methods of using death as an endpoint in sepsis studies. In summary, cfDNA levels are elevated in CLP-induced sepsis and these elevations accompany pro-inflammatory and pro-coagulant responses. NETosis may be a mechanism by which cfDNA is released and NETs may drive inflammation and coagulation in CLP sepsis. Delayed DNase administration may suppress inflammation and coagulation and may be protective in polymicrobial sepsis. In future animal sepsis studies, surrogate markers of death and a sepsis scoring system can be used in place of death as an endpoint to raise the standards in conducting ethical, humane sepsis research. / Thesis / Doctor of Philosophy (PhD)