Zugl.: Bielefeld, Univ., Diss., 2005/2006

Universiẗat, Diss.--Bielefeld, 2006. / Literaturverz. S. XIII - XXVII.

Der cGMP-Signalweg ist als einer der Hauptregulatoren von diversen Funktionen in Eukaryoten bekannt; allerdings ist seine Funktionsweise in Protozoen wenig verstanden. Im Rahmen dieser Arbeit wurde eine Guanylatcyclase, gekoppelt mit N-terminalen P4-ATPase, in intrazellulären Parasiten Toxoplasma gondii gemeldet. Eine in silico-Analyse wies auf eine Aktivierung der Guanylatcyclase durch Heterodimerisierung ihrer Cyclasedomänen hin und ermöglichte wertvolle Einsichten in mögliche Funktionen ihrer ATPase-Domäne. Dieses Protein (477-kDa) bezeichnet als TgATPaseP-GC in dieser Studie, lokalisiert in der Plasmamembran am apikalen Pol des Parasiten. TgATPaseP-GC ist unempfänglich gegenüber genetischer Deletion und seine CRISPR/Cas9 unterstützte Spaltung beendet den lytischen Zyklus von T. gondii vorzeitig. Darüber hinaus reduzierte ein Cre/loxP-vermittelter Knockdown von TgATPaseP-GC die Synthese von cGMP im Tachyzoiten und inhibierte das Parasitenwachstum aufgrund von Beeinträchtigungen Motilitäts-abhängiger Prozesse des Austretens und Eindringens. Trotz seiner zeitlich beschränkten Funktion ist TgATPaseP-GC konstitutiv während des ganzen lytischen Zyklus exprimiert, welches eine post-translationale Regulierung des cGMP-Signalweges bedingt. Nicht zuletzt impliziert das Vorhandensein von TgATPaseP-GC-Orthologen in anderen Alveolata eine divergente Umfunktionierung der cGMP-Signalwege in Protozoen. Darüber hinaus wurde ein optogenetischer Ansatz verwendet, um den cGMP-Weg durch eine photo-aktivierte Rhodopsin-Guanylat-Cyclase (RhoGC) in T. gondii zu exprimiert. Dieses System erlaubte eine kontrollierte Erhöhung von cGMP durch Licht in einer schnellen und reversiblen Weise. Die Anregung von RhoGC stimulierte signifikant die Parasitenmotilität, deren Auswirkung auch mit erhöhten Eindringen und Austreten überwacht wurde; im Gegensatz zum genetischen Knockdown von TgATPaseP-GC. Das System ermöglicht die Vermittler des cGMP-Signalwegs durch Phosphoproteomics zu identifizieren. / cGMP signaling is known as one of the master regulators of diverse functions in eukaryotes; however, its architecture and functioning in protozoans remain poorly understood. In the scope of this thesis, an exclusive guanylate cyclase coupled with N-terminal P4-ATPase was reported in an obligate intracellular parasite Toxoplasma gondii. In silico analysis indicated an activation of the guanylate cyclase by heterodimerization of its two cyclase domains and offered valuable insights into possible functions of its ATPase domain. This bulky protein (477-kDa), termed in this study as TgATPaseP-GC to reflect its envisaged multifunctionality, localizes in the plasma membrane at the apical pole of the parasite. TgATPaseP-GC is refractory to genetic deletion, and its CRISPR/Cas9-assisted disruption aborts the lytic cycle of T. gondii. Besides, Cre/loxP-mediated knockdown of TgATPaseP-GC reduced the synthesis of cGMP in tachyzoites and inhibited the parasite growth due to impairments of motility-dependent egress and invasion events. Notably, despite its temporally restricted function, TgATPaseP-GC is expressed constitutively throughout the lytic cycle, entailing a post-translational regulation of cGMP signaling. Not least, the occurrence of TgATPaseP-GC orthologs in several other alveolates implies a divergent functional repurposing of cGMP signaling in protozoans. Furthermore, an optogenetic approach was utilized to induce cGMP pathway by a photo-activated rhodopsin-guanylate cyclase (RhoGC) in T. gondii. The system enabled a light-control of cGMP elevation on crucial steps of lytic cycle in a fast, spatial and reversible manner. Excitation of RhoGC significantly stimulated the parasite motility of which impact was also monitored with an increased host-cell invasion and egress; as opposed to the genetic knockdown of TgATPaseP-GC. Having an established optogenetic system in the parasite allows to identify downstream targets of cGMP signaling via phosphoproteomic analysis.

Salinity intrusion in a river may have an adverse effect on the quality of life and can be perceived as a modern-day curse. Therefore, it is important to find technical ways to monitor and forecast salinity intrusion. In this paper, we designed a forecasting model using Simple Exponential Smoothing method (SES) which performs weekly salinity intrusion forecast in Ham Luong river (HLR), Ben Tre province based on historical data obtained from the Center for Hydro-meteorological forecasting of Ben Tre province. The results showed that the SES method provides an adequate predictive model for forecast of salinity intrusion in An Thuan, Son Doc, and Phu Khanh. However, the SES in My Hoa, An Hiep, and Vam Mon could be improved upon by another forecasting technique. This study suggests that the SES model is an easy-to-use modeling tool for water resource managers to obtain a quick preliminary assessment of salinity intrusion. / Xâm nhập mặn có thể gây tác động xấu đến đời sống con người, tuy nhiên nó hoàn toàn có thể dự báo được. Cho nên, một điều quan trọng là tìm được phương pháp kỹ thuật phù hợp để dự báo và giám sát xâm nhập mặn trên sông. Trong bài báo này, chúng tôi sử dụng phương pháp Simple Exponential Smoothing để dự báo xâm nhập mặn trên sông Hàm Luông, tỉnh Bến Tre. Kết quả cho thấy mô hình dự báo phù hợp cho các vị trí An Thuận, Sơn Đốc, và Phú Khánh. Tuy nhiên, các vị trí Mỹ Hóa, An Hiệp, và Vàm Mơn có thể tìm các phương pháp khác phù hợp hơn. Phương pháp Simple Exponential Smoothing rất dễ ứng dụng trong quản lý nguồn nước dựa vào việc cảnh báo xâm nhập mặn.