The study of benthic community structure and protein expression pattern of four benthic species in effluent areas of sewage treatment plants in Kaohsiung, Taiwan Dwi Candra Pratiwi Advisor: Dr. Li-Lian Liu Institute of Marine Biology, National Sun Yat-sen University, Kaohsiung, Taiwan, R.O.C. Abstract In order to gain better understanding on the impacts of sewage treatment plant (STP) effluent on marine environment, the present study was undertaken to investigate the structure of benthic community and protein expression patterns of 4 benthic species in STP effluent areas in Kaohsiung, Taiwan. Six sites were selected, 3 near Jhong Jhou STP, 2 near NSYSU STP and one reference site. Samples were collected in December 2009 (winter) and July 2010 (summer) by bottom trawling. In winter, temperature was 26.8oC; salinity and pH were 32. 2‰ and 8.1. In summer, environmental data were 33.7oC, 31.5‰ and 8.0, respectively. In all, 5 phyla, 31 families and 48 different species were captured in two seasons. Proteomic approach by one-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis (1D SDS-PAGE) was conducted on 3 fishes, Leiognathus splendens, Apogon fasciatus, Engyprosopon multisquama, and one crab Portunus hastatoides. Environment characteristics and structure of benthic community had no significant difference among the 6 sampling sites. Protein expression patterns based on the four examined species also indicated no significant difference among sites. The complexity of pollutants in Kaohsiung Harbor is probably the main reason affecting all sampling sites. In other words, contaminated area in Kaohsiung Harbor may cover a wide range, at least III over the present sampling distance, i.e. 8km which causes no difference in community structure and protein expression pattern among control, Jhong Jhou STP and NSYSU STP sites. Key words: 1D SDS-PAGE, Leiognathus splendens, Apogon fasciatus, Engyprosopon multisquama, Portunus hastatoides

The main objective of the study was to understand the mode of interaction of protamine (Ptm), a cationic antibacterial peptide from fish milt on the Gram negative bacterial envelope. The present study was designed to resolve the question of Ptm translocation across the seemingly impermeable Gram negative cell envelope. The Gram negative pathogen Pseudomonas aeruginosa was studied as an example of a microorganism that is Ptm-sensitive but doesn’t lyse even at bactericidal concentrations. Acquired resistance to Ptm was induced in P. aeruginosa by continuous sub-culturing in nutrient rich media containing increasing concentrations of Ptm. Alterations in bacterial surface charge, LPS composition, cell morphology and Ptm localisation on acquiring resistance were also examined. Expression of outer membrane proteins significantly decreased as P. aeruginosa acquired resistance to Ptm. OprF, the major porin in P. aeruginosa was found to be stably expressed in control, revertant (Ptm-Rev) and resistant (Ptm-Res) groups. No change in expression of efflux proteins was observed as a result of induced Ptm resistance, indicating that efflux is not among the Ptm resistance mechanisms at least in P. aeruginosa. OprM, which is part of the major efflux system (MexAB-OprM) in P. aeruginosa, was found to be down-regulated in Ptm-resistant P. aeruginosa. Another outer membrane protein down-regulated in Ptm-resistant P. aeruginosa was found to be petidyl-prolyl cis trans isomerase (PPIase) which plays a major role in proper folding and maturation of channel proteins in the outer membrane. Among the sarcosinate soluble proteins, DNA dependent RNA polymerase ? and ?’ subunits were found to be down-regulated in Ptm-resistant group indicating lower transcription levels in them. Lipopolysaccharide (LPS) from the three groups of P. aeruginosa under study was isolated and separated by SDS-PAGE. LPS composition of Ptm-Res P. aeruginosa was found to be significantly different from that of the control and Ptm-Rev but was found to be similar with that of LPS from O-antigenic mutant (A+B-, which possessed only A band structures). Comparison of the zetapotential of control, Ptm-Rev and Ptm-Res P. aeruginosa, proved that electrostatic shielding was coincidental in acquired resistance to Ptm in P. aeruginosa. The MIC of the parent strain of P. aeruginosa (A+B+) and the O-antigenic mutants (A+B-, A-B+ and A-B-) were found to be the same which may be indicating that alterations in O-antigenic components alone cannot contribute to Ptm resistance. Effects of Ptm treatment on morphologies of E. coli, S. typhimurium and P. aeruginosa whole cells and spheroplasts were also studied using transmission immuno-electron microscopy. Condensation of cytoplasmic contents was observed when whole cells and spheroplasts were treated with Ptm. Also, Ptm-treated cells and spheroplasts were stained with colloidal gold-labelled antibodies against Ptm to determine distribution within the target cells. It was quite evident that Ptm internalised in whole cells and spheroplasts without lysis and was found to be concentrated in the cytoplasm. Morphological changes observed in Ptm-Rev P. aeruginosa when exposed to Ptm were comparable with that of the control. Condensation of cytoplasmic contents was not observed in Ptm-Res P. aeruginosa when challenged with Ptm. Most of the Ptm was localized at or near the outer membrane of Ptm-treated Ptm-Res P. aeruginosa, indicating decreased outer membrane permeability. Results obtained from these experiments confirm that the resistance to Ptm observed in P. aeruginosa is at the very least, coincidental with the pleiotropic mutations involving change in outer surface including change in LPS composition, loss of porins and or alterations of porin size in OprF.

Fusarium virguliforme is a soil-borne pathogen that causes sudden death syndrome (SDS) disease in soybean. SDS is one of the most significant diseases of soybean in the United States. Fungal infection results in root and crown rot as well as SDS typical foliar symptoms including chlorosis, necrosis and complete defoliation. The use of soybean cultivars tolerant to SDS is still the most effective way to overcome the disease. On the other hand, the fungal isolates are known to have varied levels of aggressiveness on soybean indicated by the field and greenhouse experiments. Understanding the pathogen and its defense mechanism is the first step in exploring the pathogen-plant interaction. Therefore, the primary aim of this research was to elucidate the mechanism behind F. virguliforme response to soybean defense mechanisms. We further attempted to identify chromosome length polymorphism among F. virguliforme isolates and characterize the possible relationship to their level of aggressiveness. In order to fulfill the first objective, a series of differentially expressed genes were identified in F. virguliforme in the presence of soybean phytoalexin, glyceollin. The Fvgrx2 gene, a Saccharomyces cerevisiae grx2homologue, was selected for further analysis. This study demonstrates for the first time the identification and characterization of dithiol glutaredoxin gene in F. virguliforme . The role of FvGRX2 in the fungal defense to phytoalexin, glyceollin and induced oxidative burst was also investigated by generating anFvΔgrx2 knockout. In order to establish a link between the fungal karyotype and the level of fungal aggressiveness, the chromosome length polymorphism (CLP) was assessed for twenty-two F. virguliforme isolates exhibiting different levels of aggressiveness on soybean. The findings are instrumental in identifying novel pathogenicity such as the ones involved in phytotoxin production, fungicide resistance and aggressiveness.

AN ABSTRACT OF THE THESIS OF JESSE MILLER, for the Master of Science degree in PLANT, SOIL, and AGRICULTURAL SYSTEMS, presented on May 13, 2016 at Southern Illinois University Carbondale. TITLE: THE EFFECTS OF STARTER FERTILIZER ON SOYBEAN INFESTED WITH FUSARIUM VIRULIFORME OR RHIZOCTONIA SOLANI MAJOR PROFESSOR: Dr. Jason Bond Fusarium virguliforme (Aoki), the fungus that causes sudden death syndrome of soybeans (SDS), is prevalent in most of the soybean (Glycine max L. Merr.) production regions throughout the United States. Sudden death syndrome management has been limited to cultural practices and host resistance. Rhizoctonia solani (Kühn) is a fungus responsible for pre-emergence and post emergence damping off. Control methods include seed treatments and cultural practices. Several companies have advocated the use of in-furrow starter fertilizers in soybean production. Promoting root growth and emergence are a couple of the alleged benefits. It is unknown if the increased fertility in the root zone may actually increase or decrease the severity of root or seedling diseases. An objective of this study is to determine if the starter fertilizers (2-6-16), (7-12-11), (3-10-13) Nachurs Alpine Solutions™ impacts seedling disease caused by Rhizoctonia solani and soybean yield. A second objective is to determine if starter-fertilizer influences the incidence and severity of SDS and soybean yield. One trial was infested with R. solani at the rate of 0.9 g of inoculum/30.5 centimeters of row. A second trial was infested with F. virguliforme at the rate of 2.25 g/30.5 centimeters of row. Inoculum consisted of sterilized white sorghum inoculated with either pathogen. Plots were 3.04 meters wide by 6.1 meters in length with row spacing of 0.76 meters. Trials took place during the growing season of 2014 and 2015. In 2014, a randomized complete block design consisted of 4 treatments that were replicated 6 times and planted into 4 row plots. Treatments consisted of treated (Metalaxl™, Fluxapyroxad™, Pyraclostrobin™, and Imidacloprid™) or non-treated seed (‘Asgrow 4730’) combined with either fertilizer (2-6-16) or non-fertilizer. Across both trials, there were no seed treatment and fertilizer rate interactions. In the R. solani trial, stand counts were similar between the fertilizer and non-fertilizer treatments. Stand counts were higher when the seed treatment was used. There was no significant difference in soybean yield regardless of treatment. In the F. virguliforme trial, stand counts were reduced in the fertilizer treatment when compared to the non-fertilizer treatment. Foliar symptoms of SDS and soybean yield were not affected by treatment. In 2015, there were changes in treatment structure due to additions of fertilizer treatments 7-12-11 and 3-10-13. Seed treatments and randomized complete block design remained for 2015. Stand counts were higher in plots that received fertilizer treatments in the R. solani trial. Stand counts were lower in R. solani plots with treated seed. Yield was not influenced by seed treatment but was increased by 3-10-13 and 7-12-11 fertilizer treatments. For the F. virguliforme trial, reduced stand counts were found in the plots with seed treatments. Seed treatments did not influence yield. Fertilizer did not impact stand or yield. Foliar symptoms of SDS were not influenced by seed treatment or fertilizer.

Most histories of the New Left emphasize that some variant of Marxism ultimately influenced activists in their pursuit of social change. Through careful examination of Students for a Democratic Society (SDS), I argue that New Left thought was not always anti-liberal. Founding SDS members hardly rejected liberal political theory during the early years of the movement (1960-1963). New Left thought was profoundly indebted to John Dewey's political and philosophical method. Deweyan liberalism suggested theory should be directly applicable in the world of social action and truth should always be regarded as contingent. The connection between Dewey and SDS becomes apparent when one considers the role of Arnold S. Kaufman in the movement. Kaufman, a University of Michigan philosopher, applied Dewey's political and philosophical method in his work as an activist and academic. He coined the term "participatory democracy" for the New Left. Consequently, this key New Left concept was itself grounded in Dewey's philosophy.

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>Magister Scientiae - MSc / Chromosome doubling has become an important tool in breeding programmes as it offers the ability of introducing novel traits into existing plants. Doubled haploid plants are highly valued by both consumers and breeders as these plants usually show larger flower, leaves and fruit, thus making them more marketable. Marianna open pollinated plum rootstocks’ adaptability to different soil types and moisture conditions has been favoured in polyploidy studies as parental material in breeding programmes. The potential of the microtubule depolymerizing herbicide (oryzalin) for in vitro chromosome doubling were investigated by optimizing the concentration and incubation time of plant shoots to the antimitotic agent. Meristem tissues were treated for two time intervals (24 and 48 h) with five different concentrations of oryzalin (50, 75, 100, 150 or 200 μM) in liquid Murashige and Skoog (MS) medium. After treatment, plants were allowed to grow under a 16/8 h light/dark photoperiod at 24±2˚C for 4 weeks. One and two-dimensional gel electrophoresis (2-DE) coupled with mass spectrometry (MS) was used to separate, visualise and identify differently expressed proteins. Furthermore, changes in ROS accumulation, photosynthetic pigmentation, lipid peroxidation and antioxidant enzyme activity (SOD, APX and GR) were investigated. Flow cytometry results revealed that treatment of plants with oryzalin concentrations ranging from 75 to 150 μM induced ploidy after 24 h exposure whereas, 200 μM produced mixoploids containing both tetraploid and octoploids plants after 24 h exposure. Longer incubations of 48 h were detrimental to plant tissues as complete mortality was observed in the higher concentration (100 to 200 μM) treatments. Mass spectrometry analysis identified 14 differentially expressed protein spots that were characterized into different functional categories. ROS accumulation, the extent of lipid peroxidation and antioxidant capacity were differentially regulated in response to oryzalin treatment whereas photosynthetic pigments were significantly enhanced. The results suggests that oryzalin-induced proteins may act as potential biomarkers to improve fruit characteristics in future breeding programs whereas antioxidant enzymes play an important role in scavenging ROS in plants to enhance their adaptability to different environmental conditions.

Edible insects have been part of the diet of many countries across the world for several decades. It is becoming increasingly popular in the europian developed countries, mainly because of the content of nutritionally important substances. The theoretical part of the thesis deals with the characterization of edible insects in terms of nutritional benefits, explains the reasons and method of breeding edible insects. For the analysis of nutritionally active substances, flour of house cricket (Acheta domesticus) was used. A. domesticus belongs to the representatives of insects legalized from 1.1. 2018 on the EU market. Of the significantly nutritional substances, proteins, lipids, fatty acids, fiber and selected minerals were analyzed. In this study, optimalization of vertical electrophoresis SDS-PAGE was performed to determine protein fragment sizes. In the second part the influence of acricket flour addition protein bars was investigated by sensory analysis. In the experimental part that was found that cricket flour contains protein, lipids and fatty acids in significant amounts. It also contains important minerals for human health such as magnesium, potassium, iron or zinc. However, the sensory properties of protein bars for consumers were not very attractive.

This thesis describes basic principles of IP Multimedia Subsystem, known as the IMS, and shows the possibility for design applications using IMS. Firstly, a generic architecture IMS as layer model divided into four logic layer with definition the interfaces and the main entities is described. The IMS is based on SIP and IP protocols. Then is the architecture of SIP with the main entities explained. Structure of packet, request and response method in the part of SIP is also described. After that, three network services, namely presence services, instant messaging services and push over cellular services are presented. For each services the figuration of their architecture and the basic definition of their main entities are shown. Moreover, the graphical diagrams of SIP signalization are depicted for these services. In the next chapter, the self implementation of services was designed, namely presence services and services for speech transmission with next secondary function as sending email with registration information of user. The project was designed as client - server application in the development studio SDS Ericsson 4.1. The client application was simulated as java application and also as an application for Symbian emulator. The process of client registration was performed with the help of ICP platform. Finally, the diagram of analysis communication client - server is displayed with the captured packets explanation by the help of network analyzer WireShark. The video files, which were specially created for this thesis showing presentations for presence services and sending email. The most important results are summarized in the end of this article.

Submitted work describes IMS (IP Multimedia Subsystem) and focuses on services offered by this technology. In the theoretical part of the thesis is a detailed description of the IMS architecture, in the terms of its components and interfaces. Also a SIP protocol is described as the main signalling protocol, which represents the basis of the IMS architecture and RTP protocol that allows the transmission of multimedia data in real-time. Further, there are provided services that the IMS system brings. The practical part of the work describes the design and implementation of services within the IMS. It is a client-server service. Server part of the system is implemented as a SIP Servlet application. It has access to the MySQL database, which stores records of multimedia files. Client registered in the IMS network can communicate with the SIP servlet sending a request for information on such files. Following this communication, the client can play the file in his online ICP application, or download it into his PC and then play. To play and view multimedia fines is used JMF. The service is implemented in a development environment SDS Ericsson 4.1, which allows simulation of the IMS network.