Bacterial infections are a major cause of morbidity and mortality in both the community and nosocomial settings, particularly among the elderly and chronically ill. Sepsis is the body’s response to antigens and toxins released by the invasive pathogenic organisms that cause infection. When infection is not effectively controlled, sepsis may develop and progress to severe sepsis and septic shock. Early diagnosis and treatment is pivotal for survival in severe sepsis and particularly, septic shock. Our research focuses on developing a novel treatment strategy for septic shock by using single, ultra-high doses of aminoglycosides. In this project, the effect of a single, ultra-high dose of gentamicin in clearing bacteria from the blood and reducing the bacterial burden in vital organs was evaluated in a rat model of E. coli (Bort strain) induced peritonitis with severe sepsis/septic shock. Serum cytokine levels and serum lactate levels were serially measured. Further, the potential adverse effects of ultra-high dosing of aminoglycoside antibiotics in a short-term (9 h) invasive study and long term (180 days) non-invasive study were assessed. Neuromuscular paralyses due to ultra-high doses of aminoglycosides were assessed. In addition, renal injury markers such as serum creatinine and urinary Neutrophil Gelatinase Associated Lipocalin (NGAL) were assayed. The auditory and vestibular function was also assessed after ultra-high dosing of aminoglycoside in the long-term study. We conclude that animals can tolerate ultra-high doses of aminoglycosides with appropriate support. Animals were under neuromuscular paralysis for 28 – 50 minutes and were on ventilator support after single ultra-high doses (80 and 160 mg/kg) of aminoglycoside antibiotics (gentamicin and tobramycin). There was no significant acute or delayed renal or ototoxicity associated with the single, ultra-high dose aminoglycoside therapy. Histology studies of the kidneys and the cochlea of single, ultra-high aminoglycoside dosed animals and untreated control animals were performed after 180 days (6 months). Results indicated that there were no morphological differences between the treated and untreated control animals. Terminal deoxy-nucleotidyl transferase dUTP nick end labeling (TUNEL) assay of kidney tissue indicated that there was no apoptosis of endothelial cells in the tubular and glomerular regions with single, ultra- high dose of aminoglycosides consistent with an absence of ultrahigh dose induced nephrotoxicity. In the septic shock model, the E. coli Bort was below the limit of detection from the blood of the animals within minutes after single, ultra-high dose aminoglycoside administration. After necropsy, bacterial load was determined from all the vital organs and peritoneal fluid (site of infection). The bacterial levels were below the detection limit from the kidneys and there was a significant reduction in bacterial counts from all the remaining organs compared to the infected control animals. A decrease in serum cytokine and serum lactate levels compared to baseline was observed after ultra-high doses of aminoglycosides in the septic shock animals. Our studies have indicated that the ultra-high dose gentamicin is well tolerated by rats. It is highly effective in clearing E. coli Bort from the blood and reducing the bacterial burden in the organs in an experimental model of bacterial peritonitis/septic shock. Further studies in larger animals such as rabbits, sheep, pigs or dogs are required to confirm these results. If these findings are replicated in larger animals, this therapy may be developed further from ‘lab to bedside’ to treat septic shock patients in intensive care units (ICUs). / October 2015

I have decided to deal with the issue of "Risk Sources of Ammonia Cooling Systems and Their Eventual Accidental Impacts" as the risk of leakage from the cooling systems using ammonia as a coolant still prevails these days. Eventual accidental leakage simulation models were applied to several winter stadiums using ammonia as a coolant. Based on these results, it has been assessed how dangerous these sources for the citizens living in their surroundings are and measures to increase safety were suggested, too.

Made available in DSpace on 2014-07-29T16:11:52Z (GMT). No. of bitstreams: 1 Dissertacao Flaubertt 1iniciais.pdf: 86457 bytes, checksum: 527b584507a695284399e78f10a55608 (MD5) Previous issue date: 2008-03-12 / O uso de plantas medicinais tem sido muito significativo nos últimos anos, sendo incentivado pela Organização Mundial de Saúde (OMS). Synadenium umbellatum Pax, Euphorbiacea (vulgo cola-nota, cancerola, milagrosa) tem o látex usado empiricamente como anti-tumoral e antiinflamatório. Por existir espécies tóxicas nesta família e visando à segurança no uso de extratos vegetais, tal estudo avaliou a toxicidade pré-clinica do látex e do extrato etanólico das folhas (EEF) de S. umbellatum, por via oral, em ratas Wistar. O estudo seguiu diretrizes da OECD (Organisation for Economic Cooperation and Development) para teste de toxicidade aguda (Guideline 423) e de toxicidade subaguda (Guideline 407). Na toxicidade aguda do látex e do EEF, não se observou, na dose de 2000 mg/kg, alterações fisiológicas e comportamentais, e nem a morte dos animais. No entanto, o látex ocasionou congestão e infiltrado leucocitário nos rins, fígado e pulmões, efeitos não observados com o EEF. Na toxicidade subaguda, doses de 50, 100 e 200 mg/kg de EEF não produziram alterações dose-dependentes significativas nos parâmetros laboratoriais e nem alterações fisiológicas, macroscópicas e histopatológicas dos órgãos. O EEF da S. umbellatum é praticamente atóxico em exposição aguda, já o látex pode ocasionar alterações histopatológicas no fígado e pulmões. O uso crônico da planta S. umbellatium merece mais estudos.

Master of Public Health / Department of Diagnostic Medicine/Pathobiology / John A. Pickrell / Airway inflammation is characterized by the release of pro-inflammatory cytokines (IL-6) and chemokine (IL-8) from airway epithelial cells. To screen for the potential inhalation toxicity as inflammation, we tested exposure of metal oxide nanoparticles (NPs)-Titanium dioxide, Magnesium oxide, FastAct and Titanium Silicon Oxide-Manganese (TSO-Mn)-Aerogel to BEAS 2B human bronchial epithelial cells. A monolayer of cells having 80 – 90% confluence was treated with different concentrations of the NPs and feedlot dust as positive control for inflammatory processes. Releases of IL-6 and IL-8 into the culture supernatant fluid were measured by sandwich enzyme-linked immunoassay (ELISA). Characterization of NPs such as solubility and agglomeration in cell culture media were carried out to predict the effect of these properties in cellular responses. Feedlot dust increased the release of both IL-6 and IL-8 by 3 to >5 fold, suggesting an inflammatory effect while NPs did not show any effect either at increasing the dose or duration of the incubation with cell. The NPs at higher doses reduced the total IL-6 and IL-8 released, suggesting that the NPs may have bound with the cytokine and chemokine or somehow interfered with their function. The inert activity of NPs was further investigated by inspecting cell morphology, counting viable cells and assessing mitochondrial membrane potential. Concentrations at 1000 mg/L of TiO2 and 250 mg/L TSO-Mn-Aerogel could apparently limit lung epithelial cell multiplication by partially occupying the intercellular spaces, qualitatively increasing the number of cell pores and resulting in less recovered cells after 12 hours of incubation. Cells exposed to feedlot dust and titanium NPs were less viable as indicated by propidium iodide staining, but cells exposed to TSO-Mn-aerogel were more apoptotic as indicated JC-1 staining. These changes occurred at projected inhalation exposure levels > 40-100 fold above the nuisance dust level for TiO2 and permissible exposure limit for Manganese. No MgO exposures reduced apparently recovered cells to < 50% as indicated by manual hemocytometer counts (+ 15-25% variability). The lack of toxicity was most likely reflected from the high MgO solubility in the incubating media, and the relative non-toxicity of MgO.

Submitted in fulfilment of the requirements of the Degree of Master of Technology: Biotechnology, Durban University of Technology, 2010. / Photodynamic therapy is a promising treatment for cancer. It involves the combination of a photosensitizer and light of an appropriate wavelength (laser source) to cause the destruction of cancer cells. Phthalocynanines are second–generation photosensitizers with enhanced photophysical and photochemical properties. In this in vitro study the effect of aluminium (AlTSPc) or zinc (ZnTSPc) tetrasulfophthalocyanines in its inactive and active state (laser induced) on melanoma (skin cancer cells), fibroblast (healthy normal skin cells) and keratinocyte (healthy normal skin cells) cells was evaluated. For each of the cell lines approximately 3 x 104 cells/ml were seeded onto 24-well cell culture plates and allowed to attach overnight, after which cells were treated with different concentrations of AlTSPc or ZnTSPc. The photosensitizers were synthesized at Rhodes University. After 2 hrs, cells were irradiated with a diode laser at a wavelength of 672 nm and a beam diameter of 1 cm. The laser power varied between 20-30 mW and the irradiation time was calculated to deliver a light dose of 4.5 J/cm2. Post-irradiated cells were incubated for 24 hrs before cell viability was measured using the CellTiter-BlueTM Viability Assay. Also, the efficacy of the light dose and laser source used for the killing of approximately 50% of the melanoma cancer cells were investigated. AlTSPc and ZnTSPc decreased cell viability of melanoma cancer cells to approximately 50% with photosensitizer concentrations of 40 μg/ml and 50 μg/ml respectively. These photosensitizer concentrations caused a slight decrease in the percentage cell viability of fibroblast and keratinocyte cells. Results for the dark toxicity assay showed that iii both photosensitizers in the presence of high concentrations (60 μg/ml – 100 μg/ml) showed cytotoxicity effects on melanoma cancer cells in their inactive state. This was not observed in fibroblast and keratinocyte cells treated under the same experimental conditions. The optimal AlTSPc and ZnTSPc concentrations in combination with the light dose of 4.5 J/cm2 was the most efficient in killing the melanoma cancer cells with reduced killing effects on healthy normal fibroblast and keratinocyte cells when compared to other light doses (2.5 J/cm2, 7.5 J/cm2 and 10 J/cm2). The irradiation of cells photosensitized with the optimal photosensitizer concentrations with a femtosecond laser using similar laser parameters to continuous wave laser experiments resulted in a reduction in the cell viability of healthy normal fibroblast and keratinocyte cells compared to melanoma cancer cells. The presence of DNA degradation on agarose gel, morphological changes like blebbing and ultrastructural changes like nucleus condensation indicated that photodynamic therapy treated melanoma cancer cells with the optimal concentrations of AlTSPc and ZnTSPc induced cell death via apoptosis. This concludes that low concentrations of AlTSPc and ZnTSPc activated with an appropriate laser source can be used to induce cell death in melanoma cancer cells. Both AlTSPc and ZnTSPc exhibit the potential to be used as a photosensitizer in photodynamic therapy for the treatment of melanoma cancer with the occurrence of minimal damage to surrounding healthy tissue.

PURPOSE:To assess if intensity-modulated radiotherapy (IMRT) can possibly lead to improved local control and lower incidence of vision impairment/blindness in comparison to non-IMRT techniques when treating sinonasal malignancies / what is the most optimal dose constraints for the optic pathway / and the impact of different IMRT strategies on optic pathway sparing in this setting.METHODS AND MATERIALS:A literature search in the PubMed databases was conducted in July, 2012.RESULTS:Clinical studies on IMRT and 2D/3D (2 dimensional/3 dimensional) RT for sinonasal malignancies suggest improved local control and lower incidence of severe vision impairment with IMRT in comparison to non-IMRT techniques. As observed in the non-IMRT studies, blindness due to disease progression may occur despite a lack of severe toxicity possibly due to the difficulty of controlling locally very advanced disease with a dose less than or equal to] 70Gy. Concurrent chemotherapy's influence on the the risk of severe optic toxicity after radiotherapy is unclear. A maximum dose of less than or equal to] 54Gy with conventional fractionation to the optic pathway may decrease the risk of blindness. Increased magnitude of intensity modulation through increasing the number of segments, beams, and using a combination of coplanar and non-coplanar arrangements may help increase dose conformality and optic pathway sparing when IMRT is used.CONCLUSION:IMRT optimized with appropriate strategies may be the treatment of choice for the most optimal local control and optic pathway sparing when treating sinonasal malignancy.

Thesis (MScAgric (Conservation Ecology and Entomology))--Stellenbosch University, 2008. / ENGLISH ABSTRACT: Ant infestations comprising the Argentine ant Linepithema humile (Mayr), common pugnacious ant Anoplolepis custodiens (F. Smith) and cocktail ant Crematogaster peringueyi Emery are a widespread pest problem in South African vineyards. Integrated Pest Management (IPM) programmes aimed at suppressing the problematic honeydew excreting vine mealybug Planococcus ficus (Signoret) (Hemiptera: Pseudococcidae) on grapes must include ant control to optimize the effectiveness and efficacy of mealybug natural enemies. If ants are eliminated, natural enemies are able to contain mealybugs below the Economic Threshold Level (ETL). Current strategies for ant control are limited and generally include the application of long term residual insecticides that are detrimental to the environment, labour intensive to apply and can disrupt natural biological control if applied incorrectly. A more practical method of ant control using low toxicity baits was therefore investigated. Field bait preference and bait acceptance assessments aimed at determining bait repellency and palatability, respectively, were carried out during spring, summer and autumn in three vineyards of the Cape winelands region during 2007/08. Five toxicants comprising gourmet ant bait (0.5%), boric acid (0.5%), fipronil (0.0001%), fenoxycarb (0.5%) and spinosad (0.01%) dissolved in 25% sugar solution were tested against a 25% sucrose solution control. Gourmet ant bait was significantly more preferred and accepted by all ant species than the other baits. Laboratory bait efficacy assessments using four insecticides (gourmet, boric acid & spinosad) at concentrations of 0.25; 0.5; 1; 2 and 4 times the field dose and fipronil at 0.015625; 0.03125; 0.0625; 0.125; 0.25 times the field dose were carried out. Results revealed that boric acid (2%), gourmet ant bait (2%) and fipronil (1.0 X 10-5%) exhibited delayed toxicity for L. humile and C. peringueyi while spinosad (0.01%) showed delayed action on L. humile. Field foraging activity and food preference tests were also carried out for the three ant species during 2007/08. Foraging activity trials revealed that vineyard foraging activity of L. humile is higher relative to A. custodiens and C. peringueyi. This means fewer bait stations are required for effective L. humile control making low toxicity baits a more affordable and practical method of controlling L. humile than the other two ant species. Food preference trials showed that L. humile and C. peringueyi have a high preference for sugar while A. custodiens significantly preferred tuna over other baits. However, all ant species had a preference for wet baits (25% sugar water, 25% honey, tuna & agar) as opposed to dry ones (fish meal, sorghum grit, peanut butter & dog food). This research concludes that low toxicity baits show potential in ant pest management and can offer producers with a more practical, economical and environmentally friendly method of ant control which is compatible with vineyard IPM programmes. / AFRIKAANSE OPSOMMING: Mierbesmetting wat uit die Argentynse mier Linepithema humile (Mayr), die gewone malmier Anoplolepis custodiens (F. Smith) en die wipstertmier Crematogaster peringueyi Emery bestaan, is ’n plaagprobleem wat wydverspreid in Suid-Afrikaanse wingerde voorkom. Programme vir geïntegreerde plaagbeheer (GPB) wat daarop gemik is om die wingerdwitluis Planococcus ficus (Signoret) (Hemiptera: Pseudococcidae) – wat ’n probleem is weens die heuningdou wat dit afskei – op druiwe te beheer, moet mierbeheer insluit om sodoende die uitwerking en doeltreffendheid van die witluis se natuurlike vyande die beste te benut. As miere uitgeskakel kan word, sal dit vir die natuurlike vyande moontlik wees om die witluis sodanig te beheer dat dit onder die ekonomiese drempelvlakke (EDV) bly. Huidige strategieë om miere te beheer, is beperk en sluit gewoonlik die toediening van insekdoders in wat lank neem om in die grond af te breek, wat skadelik vir die omgewing is, waarvan die toediening arbeidsintensief is en wat die natuurlike biologiese beheer kan versteur indien dit verkeerd toegepas word. Daarom is ’n meer praktiese metode ondersoek waar miere deur die gebruik van lae toksisiteit lokase beheer word. Ondersoeke na lokaasvoorkeure en lokaasaanvaarbaarheid in die praktyk, wat daarop gemik is om te bepaal of die lokaas onderskeidelik afstootlik en smaaklik bevind word, is oor lente, somer en herfs in drie verskillende wingerde in die Kaapse wynlandstreek gedurende die 2007/08-seisoen uitgevoer. Vyf gifstowwe, bestaande uit gourmet ant bait (0.5%), boorsuur (0.5%), fiproniel (0.0001%), fenoksiekarb (0.5%) en spinosad (0.01%) wat in ’n 25%-suikeroplossing opgelos is, is getoets teenoor ’n kontrole wat uit ’n 25%-sukrose-oplossing bestaan. Al die mierspesies het gourmet ant bait bo die ander lokase verkies en aanvaar. In die laboratorium is ondersoeke gedoen om die doeltreffendheid van die lokase te bepaal deur vier insekdoders (gourmet ant bait, boorsuur en spinosad) te gebruik in konsentrasies van 0.25; 0.5; 1; 2 en 4 keer die dosis in die praktyk en fiproniel teen 0.015625; 0.03125; 0.0625; 0.125; 0.25 keer die dosis in die praktyk. Resultate het getoon dat boorsuur (2%), gourmet ant bait (2%) en fiproniel (1.0 X 10-5%) vertraagde toksisiteit getoon het vir L. humile en C. peringueyi, terwyl spinosad (0.01%) ’n vertraagde uitwerking getoon het op L. humile. Toetse om kossoekaktiwiteite in die praktyk en die voedselvoorkeure van die drie mierspesies te ondersoek, is ook gedurende die 2007/08-seisoen gedoen. Proewe oor kossoekaktiwiteite het getoon dat hierdie aktiwiteite in die wingerd by L. humile hoër is in verhouding met A. custodiens en C. peringueyi. Dit beteken dat minder lokaasstasies nodig is om L. humile doeltreffend te beheer en lei daartoe dat lae toksisteit lokaas ’n beter manier is om L. humile te beheer as die ander twee mierspesies. Proewe oor voedselvoorkeure het aangedui dat L. humile en C. peringueyi ’n groot voorkeur toon vir suiker, terwyl A. custodiens ’n duidelike voorkeur vir tuna het. Alle mierspesies het egter ’n voorkeur vir nat lokaas (25% suikerwater, 25% heuning, tuna en agar), eerder as droë lokaas (vismeel, sorghumgruis, grondboontjiebotter en hondekos) getoon. Uit hierdie navorsing word afgelei dat lae toksisteit lokaas potensiaal toon in mierbeheer en dat dit produsente ’n meer praktiese, ekonomiese en omgewingsvriendelike metode van mierbeheer kan bied wat met GPB-programme in die wingerd versoenbaar is.

The development of a highly sensitive ruthenium-based fluorescent staining solution isdescribed in this dissertation. The in-house synthesized ruthenium complex (RuMS)containing both sulfonated and non-sulfonated ligand has detection limit of 1 ng ofprotein that is better than colloidal coomassie, silver and ruthenium complex containingall sulfonated ligands (RuBPS). RuMS stain has 100-fold dynamic range and does notinterfere with subsequent mass spectral identification of proteins. The capability of inhousesynthesis of the staining solution makes it a viable cost-effective alternative to theexpensive commercially available fluorescent stain, Sypro Ruby. The low detection limit,broad linear dynamic range and compatibility with mass spectrometry, make thedevelopment of this stain a worthwhile pursuit. The staining solution was utilized insubsequent applications of two-dimensional gel electrophoresis (2-DE) technology.Proteomics methodology utilizing 2-DE and mass spectrometry was applied toinvestigate the effect of malathion on the proteome of human neuroblastoma cells.Results indicated that out of 122 proteins that were identified from the neuroblastomaproteome, sixteen proteins were down-regulated while five proteins were significantlyup-regulated after treatment with malathion. Significant down-regulation of calciummodulators like calmodulin and calgizarrin and other key chaperones makes themalathion-treated cells highly prone to oxidative stress. With increased awareness inpesticide related adverse effects, identification of altered proteins in malathion-treatedhuman neuroblastoma cells is a critical finding.Proteomics is a major area of research in the identification of biomarkers for diseases. Anovel immunoprecipitation method developed in this work allowed for successfulisolation and identification of albumin-interactome in cerebrospinal fluid (CSF) that isusually under-represented in standard CSF analysis using 2-DE. A key finding is thedifferential expression of various isoforms of proteins in CSF albumin-interactome fromAlzheimer's disease (AD) subjects. The data implicate the acidic isoform ofprostaglandin D2 synthase (PGDS2) as a potential biomarker for AD. An understandingof the differential expression of these protein isoforms in AD will provide insight into theetiology of the disease and this can have far-reaching implication on drug developmentleading to the cure or even preventation of the disease.

The increasing urbanization of landscapes significantly alters the surface water hydrology of impacted watersheds. As a side effect, stormwater discharges to receiving water bodies are often of decreased quality due to pollutants deposited on impervious urban surfaces being entrained by runoff. A pertinent example of this problem is the presence of copper and zinc in highway runoff. Both copper and zinc have been shown to exert toxic effects on aquatic micro- and macro-biota. Copper in particular has been shown to harmfully disrupt the olfactory nervous system of fish species at concentrations as low as 3 [mu]g/L. To meet these limits, treatment of highway runoff for the removal of copper and zinc is necessary. However, due to the complexities associated with the behavior of heavy metals in natural systems, the appropriateness of removal techniques will necessarily depend on a variety of system-specific factors and chemical characteristics of highway runoff. Adsorption has been shown to be generally effective in the removal of dissolved heavy metals, but the choice of adsorptive media is again dependent on system-specific parameters. This study developed and evaluated a column testing protocol that can be used to quickly and reliably evaluate adsorptive removal of dissolved heavy metals from highway runoff. The protocol is demonstrated in an evaluation of iron oxide, manganese oxide, crab shell, concrete, and bone meal media for removing dissolved copper and zinc from highway runoff. The performance of these media was assessed as a function of various runoff characteristics including pH, ionic strength, alkalinity, and total organic carbon. The methodology was used to show that iron oxide media in combination with crab shell or concrete media provided the most effective removal of copper and zinc from highway runoff. Through this study, the convenience, flexibility, and robustness of the proposed protocol are compellingly established. / text

<p>Irrigation with saline waters is an agricultural practicethat is becoming increasingly common as competition for freshwater increases. In this thesis the mechanisms behind salinityand drought stress has been studied using data from fieldexperiments in combination with a modelling tool, theCoupModel. Measurements from field experiments on salinity,boron toxicity and drought stressed tomatoes grown during twoclimatically different seasons in the Arava desert, Israel,showed a linear relationship between relative growth andevapotranspiration, for all treatments and seasons. Data fromthe spring was used to concurrently simulate growth andtranspiration, hence accounting for feedback mechanisms betweenthe plant and the environment. Salinity stress was modelled asan osmotic effect (reduction of water uptake at high soilsalinities, W approach) or a toxicity effect (direct reductionof photosynthesis with soil salinity, G approach). Goodagreement between simulated growth and transpiration wasachieved with both salinity stress approaches, with twoexceptions. When growth and transpiration were simulated withthe W approach at different salinity levels, transpiration wasunderestimated at high stress. The G approach resulted in anunderestimation of growth at high water stress under moderatesalinity. A direct decrease of photosynthesis leads to adecreasing water-use efficiency with salinity while water-useefficiency remains constant with salinity when the salinitystress is modelled as a reduction in water uptake. Measurementsshowed decreasing water-use efficiency for the salinitygradient, explaining why the W approach was not applicable. Itwas not possible to detect any considerable differences betweenthree different approaches for water uptake tested in thestudy.</p><p><b>Keywords:</b>Water-use efficiency; osmotic effect; iontoxicity; semi-arid.</p>