Introdução: A Doença do Refluxo Gastroesofágico (DRGE) possui elevada prevalência e morbidade. O tratamento clínico consiste em recomendações quanto ao estilo de vida e, essencialmente, no uso de inibidores da bomba protônica (IBP). A aderência (Ad) dos pacientes com DRGE à prescrição, embora fundamental para o sucesso terapêutico, tem sido pouco estudada. Objetivo: Avaliar a Ad ao tratamento com IBP e possíveis variáveis relacionadas em pacientes com DRGE. Métodos: Estudo transversal e prospectivo com 240 pacientes adultos consecutivos com DRGE erosiva e não-erosiva (ne-DRGE) que haviam recebido dose padrão ou dose dobrada de omeprazol em uso contínuo. Todos os pacientes foram classificados conforme o grau, segundo os achados da endoscopia digestiva alta (EDA) em ne-DRGE (162; 67,5%), classificação endoscópica de Los Angeles (LA) A (48; 20,0%), LA B (21; 8,6%), LA C (1; 0,4%), LA D (1; 0,4%) e Barrett (7; 2,9%). Foi aplicado o questionário de Morisky et al constituído de 4 questões com respostas dicotômicas para avaliar a Ad, classificando-a como baixa (0-2 pontos) e alta ad (3-4 pontos). Foi também aplicado o questionário QS-DRGE para a avaliação dos sintomas, com 10 questões com escores de 0 a 50, conforme a frequência menor ou maior dos sintomas. Os resultados de Ad foram relacionados com os dados sócio-demográficos, polifarmácia (PF) (uso de mais de cinco medicamentos diariamente), comorbidades (CM), tempo de tratamento (TT), escore QS-DRGE, presença de sintomas descrita em prontuário, achados da EDA e conhecimento do paciente sobre a doença. Resultados: (1) 126 pacientes (52,5%) apresentaram alta Ad e 114 (47,5%) baixa Ad; (2) Os pacientes mais jovens (p = 0,002) foram menos aderentes; (3) Pacientes sintomáticos de acordo com relato em prontuário apresentaram maior percentual de baixa Ad e 2 vezes maior probabilidade de ter baixa Ad em relação aos assintomáticos (p = 0,02); (4) Os pacientes casados apresentam probabilidade 2,41 vezes maior de ter baixa Ad do que os viúvos. (5) As demais variáveis estudadas não influenciaram a Ad ao tratamento. Conclusões: Pacientes em uso de IBP em tratamento ambulatorial em hospital terciário em São Paulo apresentaram grande percentual de baixa Ad ao tratamento, sendo esta uma possível causa da falha da terapia com IBP. Idade < 60 anos e estado civil casado podem ser fatores de risco para a baixa Ad / Introduction: The Gastroesophageal Reflux Disease (GERD) is a highly prevalent disease and a major cause of morbidity. Clinical treatment is based on lifestyle recommendations and, essentially, in the use of a proton pump inhibitor (PPI). Adherence (Ad) of GERD patients to the prescribed treatment, although critical for therapeutic success, has been little studied. Objective: Assess adherence to the PPI treatment and potential associated variables in patients with GERD. Case studies and Methods: Transversal and prospective study with 240 consecutive adult patients, diagnosed with erosive GERD (e-GERD) and non-erosive GERD (ne-GERD) for whom continuous use of the standard dose or the double dose of omeprazol had been prescribed. Patients were ranked according to the findings of high digestive endoscopy (HDE) in ne-GERD (162; 67.5%); e-GERD: Los Angeles (LA) endoscopic classification A (48; 20.0%), LA B (21; 8.6%), LA C (1; 0.5%), and LA D (1; 0.5%) and Barretts esophagus (7; 2.9%). The Morisky questionnaire, that includes four questions with dicotomic responses to assess Ad, was applied. Ad was classified as low (0-2 points) and high (3-4 points). In addition, the QS-GERD questionnaire was applied to assess symptoms, using 10 questions with score 0 to 50, according to the greater or lesser symptom frequency. Ad results were correlated with personal data (gender and age), demography, polypharmacy (PF), comorbidities (CM), treatment time (TT), QS-GERD scores, symptoms described in the patients record, HDE findings and patient awareness about the disease. Results: (1) 126 patients (52.5%) exhibited high Ad and 114 (47.5%) low Ad; (2) younger patients (p = 0,002) were less compliant; (3) married patients had a 2.41 greater probability to exhibit low Ad as compared to widowers (p = 0.03); (4) patients with symptoms indicated in the patients record exhibited a lower Ad rate and twice greater probability of exhibiting low Ad as compared to asymptomatic patients (p = 0.02); (5) the other variables studied had no influence on treatment adherence. Conclusion: Patients using PPI as out-patients in third care hospital in São Paulo exhibited high rate of reduced treatment adherence, and this may be a potential cause of PPI therapy failure. Age < 60 years and marital status may be risk factors for low adherence

Proton pump inhibitors (PPIs) are commonly coadministered with clopidogrel, an antiplatelet agent, to patients with acute coronary syndrome (ACS). Mechanistic studies suggest that PPIs have the potential to competitively inhibit the bioactivation of clopidogrel and may attenuate its antiplatelet action in the body. The clinical implications of this drug-drug interaction have been extensively studied; however reported findings are inconsistent. More recently, several studies have questioned whether PPIs are associated with adverse cardiovascular events independent of clopidogrel. Given that PPIs and clopidogrel are widely used, it is critical to better understand the clinical impact of the concomitant treatment with both drugs. This thesis includes four studies that investigate the clinical effects of the drug-drug interaction between clopidogrel and PPIs. Chapter 2, a systematic review and meta-analysis, summarizes findings from 118 studies. Findings do not provide strong evidence for an association between adverse cardiovascular events and the use of PPIs when used alone, in combination with clopidogrel, or in combination with other antiplatelets. Chapters 3, 4, and 5 present analyses of real-world data comprised of electronic medical records. Results of these analyses demonstrate 1) that the concomitant use of clopidogrel and PPIs among inpatients was consistent with clinical guidelines suggested by the FDA (Chapter 3); 2) a lack of association between PPI use vs nonuse and four adverse cardiovascular outcomes among clopidogrel users (Chapter 4); and 3) a lack of association between PPI use vs nonuse and adverse cardiovascular outcomes among prasugrel users or ticagrelor users (Chapter 5). Collectively, our findings do not provide evidence of an elevated risk of adverse cardiovascular outcomes with the combined use of PPIs and clopidogrel. Although pharmacodynamic and pharmacokinetic studies have demonstrated an interaction between these two drugs, our findings support the opinion that the biological interaction does not translate into adverse clinical events among patients with acute coronary syndrome.

Long-term use of proton pump inhibitors (PPIs) is common in patients with muscle wasting-related chronic diseases. We explored the hypothesis that the use of PPIs may contribute to a reduction in muscle mass and function in these patients. Literature indicates that a PPI-induced reduction in acidity of the gastrointestinal tract can decrease the absorption of, amongst others, magnesium. Low levels of magnesium are associated with impaired muscle function. This unwanted side-effect of PPIs on muscle function has been described in different disease backgrounds. Furthermore, magnesium is necessary for activation of vitamin D. Low vitamin D and magnesium levels together can lead to increased inflammation involved in muscle wasting. In addition, PPI use has been described to alter the microbiota’s composition in the gut, which might lead to increased inflammation. However, PPIs are often provided together with nonsteroidal anti-inflammatory drugs (NSAIDs), which are anti-inflammatory. In the presence of obesity, additional mechanisms could further contribute to muscle alterations. In conclusion, use of PPIs has been reported to contribute to muscle function loss. Whether this will add to the risk factor for development of muscle function loss in patients with chronic disease needs further investigation.

Indiana University-Purdue University Indianapolis (IUPUI) / Fatty acid synthase (FASN) is over-expressed in many cancers, and novel inhibitors that target FASN may find use in the treatment of cancers. It has been shown that orlistat, an FDA approved drug for weight loss, inhibits the thioesterase (TE) of FASN, but can be hydrolyzed by TE. To understand the mechanisms of TE action and for designing better FASN inhibitors, I examined the mechanism of orlistat hydrolysis by TE using molecular dynamics simulations. I found that the hexyl tail of orlistat undergoes a conformational transition, destabilizing a hydrogen bond that forms between orlistat and the active site histidine. A water molecule can then hydrogen bond with histidine and become activated to hydrolyze orlistat. These findings suggest that rational design of inhibitors that block hexyl tail transition may lead to a more potent TE inhibitor. To search for novel inhibitors of TE, I performed virtual DOCK screening of FDA approved drugs followed by a fluorogenic assay using recombinant TE protein and found that proton pump inhibitors (PPIs) can competitively inhibit TE. PPIs, which are used for the treatment of gastroesophageal reflux and peptic ulcers, work to decrease gastric acid production by binding irreversibly with gastric hydrogen potassium ATPase in the stomach. Recently, PPIs have been reported to reduce drug resistance in cancer cells when used in combination with chemotherapeutics, although the mechanism of resistance reduction is unknown. Further investigation showed that PPIs are able to decrease FASN activity and cancer cell proliferation in a dose-dependent manner. These findings provide new evidence that FDA approved PPIs may synergistically suppress cancer cells by inhibiting TE of FASN and suggests that the use of PPIs in combinational therapies for the treatment of many types of cancer, including pancreatic cancer, warrants further investigation.

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CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / A busca por estratÃgias de cultivo que possam contribuir para a aclimataÃÃo de plantas à salinidade à de fundamental importÃncia, pois, alÃm de possibilitar a identificaÃÃo de genes potenciais para guiar ensaios de modificaÃÃo genÃtica, permite selecionar cultivares com maior capacidade de crescer em solos com excesso de sais. A fim de testar a hipÃtese que a nutriÃÃo nitrogenada com NH4+ aumenta a tolerÃncia de plantas de Sorghum bicolor à salinidade, atravÃs da ativaÃÃo de mecanismos voltados ao controle da homeostase iÃnica, estabeleceram-se trÃs etapas experimentais. Na primeira delas, que objetivou definir a relaÃÃo entre as fontes de nitrogÃnio (N), NO3- e NH4+, que proporcionasse melhor crescimento das plantas sob salinidade, observou-se claramente que a nutriÃÃo somente com NH4+ (proporÃÃo NO3-/NH4+ de 0:100) foi mais vantajosa para o crescimento de S. bicolor sob salinidade que a nutriÃÃo apenas com NO3- ou com o regime misto desses dois Ãons, dado os maiores Ãndices de Ãrea foliar e massa seca da parte aÃrea. Verificou-se tambÃm que, sob estresse, as plantas nutridas somente com NH4+ acumularam menos Na+ nas folhas e nas raÃzes, influenciando positivamente a relaÃÃo K+/Na+, e apresentaram maiores teores de aminoÃcidos solÃveis, principalmente aqueles ricos em N (glutamina e asparagina), que contribuÃram para evitar a toxicidade do NH4+ e provavelmente para o ajustamento osmÃtico. AlÃm disso, enquanto plantas nutridas com proporÃÃes NO3-/NH4+ de 100:0, 75:25, 50:50 e 25:75 apresentaram taxas de assimilaÃÃo lÃquida de CO2 inalteradas ou reduzidas pela salinidade, plantas cultivadas somente com NH4+ (proporÃÃo 0:100) apresentaram incrementos nessa variÃvel, em reposta ao estresse. A segunda etapa teve como objetivo principal investigar se a tolerÃncia à salinidade mediada pelo NH4+ era resultante da regulaÃÃo efetiva dos processos relacionados à fotossÃntese. Nessa ocasiÃo, esse argumento foi refutado, pois a melhor eficiÃncia do fotossistema II sob estresse salino foi observada nas plantas cultivadas com a mesma proporÃÃo de NO3- e NH4+ (proporÃÃo 50:50). Nesse grupo de plantas, a reduÃÃo no quenching nÃo fotoquÃmico (NPQ) confirmou a maior eficiÃncia fotoquÃmica, dado o aumento na eficiÃncia quÃntica potencial (Fv/Fm) e efetiva (&#934;PSII) do fotossistema II e a elevada taxa de transporte de elÃtrons (ETR). Esse fenÃmeno foi diretamente relacionado com os incrementos nos teores de clorofila b e de antocianinas. Por fim, na terceira etapa, objetivou-se elucidar os mecanismos envolvidos no controle do acÃmulo de Na+, sob salinidade, na cÃlula e na planta inteira, bem como identificar o papel da nutriÃÃo com NH4+ nesses processos. Em estudos com vesÃculas de membrana de raÃzes, verificou-se que plantas estressadas cultivadas somente com NH4+ apresentaram maior ativaÃÃo dos transportadores do tipo antiporte Na+/H+ (SOS1) de membrana plasmÃtica e, em menor proporÃÃo, do antiporte Na+/H+ (NHX) de tonoplasto, ao passo que o oposto foi observado nas plantas nutridas com NO3-. Esses dados sugerem que o cultivo somente com NO3- induziu o mecanismo de compartimentaÃÃo de Na+ no vacÃolo, como evidenciado pela anÃlise dos transcritos da famÃlia NHX, em que a expressÃo do gene SbNHX2 (principal isoforma expressa) nas raÃzes das plantas foi aumentada em quase todos os tempos analisados (24, 48, 120 e 240 horas apÃs exposiÃÃo ao NaCl). Mesmo assim, essa resposta nÃo foi suficiente para o controle do Na+, jà que a entrada contÃnua desse Ãon no xilema radicular afetou o influxo de K+ na seiva e limitou o acÃmulo de K+ nas folhas. Por outro lado, a nutriÃÃo somente com NH4+ ativou potencialmente mecanismos de controle do acÃmulo de Na+, uma vez que houve acionamento efetivo do efluxo de Na+ para o apoplasto via SOS1, que restringiu o carregamento desse Ãon no xilema e, consequentemente, limitou a acumulaÃÃo dele nos tecidos aÃreos. A formaÃÃo do gradiente de potencial eletroquÃmico, essencial para a atividade dos transportadores Na+/H+, foi modulada diferencialmente pela fonte de N. A atividade de bombeamento de prÃtons da H+-ATPase de membrana plasmÃtica (P-ATPase) foi estimulada em maior proporÃÃo pela presenÃa de NH4+, sem haver, contudo, aumento na atividade de hidrÃlise de ATP. Jà o aumento da translocaÃÃo de H+ pela P-ATPase em plantas estressadas cultivadas com NO3- foi diretamente relacionado ao incremento na hidrÃlise de ATP. Esses resultados sugerem que a disponibilidade de NH4+ aumentou a afinidade da P-ATPase por H+, pois houve melhor eficiÃncia de acoplamento H+/ATP, e isso tornou a enzima mais efetiva para transportar H+ com menos gasto de energia. AlÃm disso, esse aumento no bombeamento de prÃtons resultou em um maior potencial eletroquÃmico, e favoreceu diretamente a atividade do antiporte SOS1 de membrana plasmÃtica. Os nÃveis de transcritos dos genes SbPHA3 e SbPHA5 (principais isoformas expressas da famÃlia) foram aumentados nas plantas cultivadas somente com NO3-, nos tempos iniciais de exposiÃÃo ao estresse salino (12 e 24 h), enquanto que, nos cultivos somente com NH4+, essa resposta sà foi detectada apÃs 24 h. No vacÃolo, a principal bomba responsÃvel pela formaÃÃo do gradiente de H+ durante a exposiÃÃo ao estresse salino foi a H+-ATPase (V-ATPase), em comparaÃÃo à H+-PPiase. Nas plantas cultivadas somente com NO3-, observou-se uma melhor regulaÃÃo da V-ATPase, em associaÃÃo à atividade aumentada do antiporte NHX, enquanto que no cultivo com NH4+, a ativaÃÃo do transporte de H+ sob salinidade foi diretamente relacionada a incrementos na atividade de hidrÃlise de ATP da V-ATPase, bem como ao aumento da expressÃo dos transcritos do gene SbVHA2, ao longo de todo o perÃodo experimental. Essas observaÃÃes revelam que o NH4+, como fonte Ãnica de N, ativa mecanismos que envolvem uma regulaÃÃo coordenada, nas raÃzes, da atividade e da expressÃo gÃnica de bombas de H+ e transportadores Na+/H+ de membrana plasmÃtica e de tonoplasto, que culminam no controle do acÃmulo de Na+ na planta inteira e aumentam a tolerÃncia de S. bicolor ao estresse salino. / Over the last decades, several researchers have focused the development of cultivation strategies in order to improve the plantâs ability to withstand salinity. Understanding the plant salt tolerance is one of important trait to enhance productivity of crops in saline soils because it provides molecular basis for plant breeding, as well as allows identify plant species with a greater ability to grown in salinized areas. In order to test the hypothesis that nitrogen nutrition with NH4+ improves the salt tolerance in Sorghum bicolor plants, through the restrict control of ionic homeostasis, three experimental steps were established. In the first one, we investigated what would nitrogen regime, as NO3-:NH4+ ratio, contribute to the better growth of plants under salinity. Our data clearly showed that the nutrition with only NH4+ (NO3-/NH4+ at 0:100) was more advantageous for the growth of S. bicolor under salinity than the supply with solely NO3- or the mixed regimes, as evidenced by the higher leaf area and shoot dry mass. Under salinity, Na+ accumulation was severely limited in presence of NH4+ (0:100), which positively influenced on K+/Na+ homeostasis. In parallel, NH4+-fed plants displayed a substantial accumulation of N-rich amino acids (mainly glutamine and asparagine) in both tissues, which seems to be fundamental in alleviating the NH4+ toxicity. Furthermore, whereas plants treated with NO3-:NH4+ ratio of 100:0, 75:25, 50:50 and 25:75 ratios had their photosynthetic rates (A) unaltered or reduced by salinity, plants supplied with only NH4+ showed an increased CO2 assimilation in response to stress. During the second step, we evaluated if the better salt tolerance in NH4+ cultivated plants was due to an effective regulation of photosynthesis-related processes. This idea was rejected because of the most striking effects of nitrogen regime were observed in plants supplied with equal amounts of NO3-: NH4+ (50:50). Under salt stress, plants from 50:50 NO3-:NH4+ treatments displayed a lower non-photochemical quenching (NPQ) and an improved photosystem II maximum efficiency (Fv/Fm). Their superior performance was also indicated by a higher effective quantum yield of PSII (&#934;PSII) and electron transport rate (ETR), as well as increased chlorophyll b and anthocyanins. Finally, at the third step, we supply S. bicolor plants with NO3- or NH4+ to investigate changes in pathways for control of Na+ accumulation, at cell and whole plant level, in response to 75 mM NaCl-stress. By using root membrane-enriched vesicles, it was found that a more pronounced plasma membrane Na+/H+ antiporter (SOS1) activity and low loading of Na+ in the xylem in the NH4+ treated plants, whereas a largest vacuolar Na+/H+ exchanger (NHX) activity was noticed by NO3- grown plants. These data suggest that the NO3- availability induced the compartmentalization of Na+ into the vacuole, as supported by the upregulation of SbNHX2 gene expression over time of NaCl exposure (12, 24, 48, 120 and 240 h). Nonetheless, it composed an inefficient pathway of Na+ control, since the incessant entrance of Na+ in the xylem sap impaired the K+ loading and limited the K+ accumulation in the shoot. On the other hand, the NH4+ supply potentially activated the mechanisms for control of Na+ accumulation, driving an effective efflux of Na+ out of the cell, via SOS1, restricting its loading in the xylem and thus limiting Na+ reach and accumulation in the aerial tissues. Surprisingly, we found that the generation of electrochemical potential gradient for Na+/H+ exchange activity is differentially modulated by the nitrogen source. The H+-pumping activity driven by plasma membrane H+-ATPase (P-ATPase) was greatly stimulated by the presence of NH4+ in growth medium, however, without an increase in ATP hydrolysis activity. Conversely, the improvement of P-ATPase-generated H+-pumping of NO3- fed stressed plants was directly related to the increase of ATP hydrolysis. These data show that the NH4+ availability enhances the H+/ATP coupling efficiency of P-ATPase, i. e. the enzyme displayed a high capacity of transport H+ across plasma membrane with low ATP consumption. Moreover, the bigger H+ translocation resulted in a greater electrochemical potential which in turn favored the SOS1 activity. The expression of SbPHA3 and SbPHA5 genes was upregulated in NO3- grown stressed plants at the beginning of salt exposure (12 and 24 h), whereas it was enhanced in NH4+ supplied stressed plants only after 24 h. At vacuole level, the H+-ATPase (V-ATPase) was the main proton pump responsive to salinity, as compared do H+-Pyrophosphatase (PPase). A better regulation between V-ATPase and NHX antiporter activities was noticed by plants from NO3- treatments. Under NH4+ supply, the increase of H+ pumping was directly associated to the improvement of ATP hydrolysis by V-ATPase, coupled to upregulation of SbVHA2 gene expression over time of salinity exposure. Taken together, our data reveal that the NH4+, as the only nitrogen source, activates an intricate regulation of Na+ control pathways, involving the existence of a robust regulation and systematic mechanism firstly on root cell and subsequently on whole plant in sorghum upon salinity. In conclusion, the NH4+ stimulated salt tolerance is resulted from a more active SOS1 protein and high efficiency of P- and V-ATPase in the roots, which help to efficient Na+ exclusion and counteract net Na+ accumulation in the cytosol, thus preventing the loading of Na+ in the xylem sap and its reach in the photosynthetic tissues.

Die arbuskuläre Mykorrhiza (AM) ist eine mutualistische Symbiose, die die Phosphataufnahme und Pathogenresistenz von Pflanzen verbessern kann. In der vorliegenden Doktorarbeit wurde die Rolle der Protonen-pumpenden ATPase MtHA1 für die AM Symbiose in Medicago truncatula untersucht. In MtHA1 Mutanten konnten AM Pilze nur noch verkürzte Arbuskel ohne typische Verzweigungen ausbilden. Dies zeigte sich auch in Expressionsmustern von Genen, die für Proteine in verschiedenen Bereichen der periarbuskulären Membran kodieren. Außerdem waren AM Pilzbesiedelung, die verbesserte Nährstoffaufnahme und die Wachstumsförderung in MtHA1 mutierten Pflanzen reduziert. Die Mykorrhiza-induzierte Resistenz (MIR) wurde näher in M. truncatula Pflanzen untersucht, die von Aphanomyces euteiches infiziert waren, dem Erreger einer Wurzelfäule in Leguminosen. In einem geteilten Wurzelsystem, das eine hohe Expression von Verteidigungsgenen aufwies, unterdrückte ein AM Pilz diese Expression und erhöhte in Folge die Empfindlichkeit für das Pathogen. In Wurzeln von Topfkulturen dagegen konnte eine typische MIR beobachtet werden, die wahrscheinlich auf erhöhter Aktivität der Jasmonat/Ethylen-regulierten Verteidigungsantwort beruht, verursacht durch eine Unterdrückung der Salizylsäuresynthese. Im Ergebnis zeigt diese Arbeit die bedeutende Rolle des Gens MtHA1 für die Bildung und Funktion der arbuskelhaltigen Zellen. Die Mutation des Gens führt zur verminderten Arbuskelverzweigung, reduzierter Phosphataufnahme und Wachstumsförderung in der Mykorrhiza und schließlich zu einer geringeren Gesamtbesiedelung durch den AM Pilz. Genexpressionsanalysen weisen darauf hin, dass unterschiedliche Mechanismen den lokalen und systemischen Wechselwirkungen zwischen AM Pilzen und Pathogenen in der Wurzel zu Grunde liegen. Verschieden physiologische Zustände von geteilten Wurzelsystemen und Wurzeln in Topfkulturen erschweren allerdings einen direkten Vergleich der beiden experimentellen Ansätze. / Arbuscular mycorrhiza (AM) is a wide spread mutualistic symbiosis, which can improve phosphate acquisition and pathogen resistance of plants. In the current Ph.D. thesis the role of a proton pumping ATPase (MtHA1) for the AM symbiosis in Medicago truncatula was investigated. In MtHA1 mutant plants, different AM fungi only developed truncated arbuscules without forming typical hyphal branches, and this phenotype was mirrored by expression patterns of genes for proteins located in different areas of the periarbuscular membrane. AM fungal colonization, improved phosphate uptake and plant growth promotion were reduced in MtHA1 mutant plants. Mycorrhiza-induced resistance (MIR) and the nodule symbiosis were, however, not affected. MIR was further analyzed in the M. truncatula infected with Aphanomyces euteiches which causes a root-rot disease in legumes. In a split root system showing high levels of defense-gene expression, colonization of an AM fungus reduced this expression and in consequence increased susceptibility of the roots for the pathogen. In roots of pot cultures, however, a typical MIR was observed and could be based on the higher activity of jasmonate/ethylene-regulated defense responses due to suppression of salicylic acid biosynthesis. In conclusion, this work shows that the gene MtHA1 encoding a proton pumping-ATPase plays a critical role in the formation and function of arbuscule-containing cells. Expression of the mutated gene results in reduced formation of arbuscule branches. This in turn negatively influences mycorrhizal phosphate uptake, plant growth promotion and overall mycorrhizal colonization of the roots. Gene expression analyses indicate that different mechanisms underlay local and systemic interactions between the mycorrhizal fungus and the root pathogen. The different physiological stages of pot culture and split root system make a comparison of the two experimental approaches, however, difficult.

Thesis (M.Pharm. (Pharmacy Practice))--North-West University, Potchefstroom Campus, 2008.