Regulation of Rab5 GTPase activity during Pseudomonas aeruginosa-macrophage interaction

Pseudomonas aeruginosa is a Gram-negative opportunistic pathogen. Several antibiotic resistant strains of P. aeruginosa are commonly found as secondary infection in immune-compromised patients leaving significant mortality and healthcare cost. Pseudomonas aeruginosa successfully avoids the process of phagocytosis, the first line of host defense, by secreting several toxic effectors. Effectors produced from P. aeruginosa Type III secretion system are critical molecules required to disrupt mammalian cell signaling and holds particular interest to the scientists studying host-pathogen interaction. Exoenzyme S (ExoS) is a bi-functional Type III effector that ADP-ribosylates several intracellular Ras (Rat sarcoma) and Rab (Response to abscisic acid) small GTPases in targeted host cells. The Rab5 protein acts as a rate limiting protein during phagocytosis by switching from a GDP- bound inactive form to a GTP-bound active form. Activation and inactivation of Rab5 protein is regulated by several Rab5-GAPs (GTPase Activating Proteins) and Rab5-GEFs (Rab5-Guanine nucleotide Exchange Factors). Some pathogenic bacteria have shown affinity for Rab proteins during infection and make their way inside the cell. This dissertation demonstrated that Rab5 plays a critical role during early steps of P. aeruginosa invasion in J774-Eclone macrophages. It was found that live, but not heat inactivated, P. aeruginosa inhibited phagocytosis that occurred in conjunction with down-regulation of Rab5 activity. Inactivation of Rab5 was dependent on ExoS ADP-ribosyltransferase activity, and more than one arginine sites in Rab5 are possible targets for ADP-ribosylation modification. However, the expression of Rin1, but not other Rab5GEFs (Rabex-5 and Rap6) reversed this down-regulation of Rab5 in vivo. Further studies revealed that the C-terminus of Rin1 carrying Rin1:Vps9 and Rin1:RA domains are required for optimal Rab5 activation in conjunction with active Ras. These observations demonstrate a novel mechanism of Rab5 targeting to phagosome via Rin1 during the phagocytosis of P. aeruginosa. The second part of this dissertation investigated antimicrobial activities of Dehydroleucodine (DhL), a secondary metabolite from Artemisia douglasiana, against P. aeruginosa growth and virulence. Populations of several P. aeruginosa strains were completely susceptible to DhL at a concentration between 0.48~0.96 mg/ml and treatment at a threshold concentration (0.12 mg/ml) inhibited growth and many virulent activities without damaging the integrity of the cell suggesting anti-Pseudomonas activity of DhL.

U.S. Army front line artillery observation during the 1945 Okinawa campaign: An oral history case study

This dissertation explores the role of artillery forward observation teams during the battle of Okinawa (April–June 1945). It addresses a variety of questions associated with this front line artillery support. First, it examines the role of artillery itself in the American victory over the Japanese on Okinawa. Second, it traces the history of the forward observer in the three decades before the end of World War II. Third, it defines the specific role of the forward observation teams during the battle: what they did and how they did it during this three-month duel. Fourth, it deals with the particular problems of the forward observer. These included coordination with the local infantry commander, adjusting to the periodic rotation between the front lines and the artillery battery behind the line of battle, responding to occasional problems with "friendly fire" (American artillery falling on American ground forces), dealing with personnel turnover in the teams (due to death, wounds, and illness), and finally, developing a more informal relationship between officers and enlisted men to accommodate the reality of this recently created combat assignment. Fifth, it explores the experiences of a select group of men who served on (or in proximity to) forward observation teams on Okinawa. Previous scholars and popular historians of the battle have emphasized the role of Marines, infantrymen, and flame-throwing armor. This work offers a different perspective on the battle and it uses new sources as well. A pre-existing archive of interviews with Okinawan campaign forward observer team members conducted in the 1990s forms the core of the oral history component of this research project. The verbal accounts were checked against and supplemented by a review of unit reports obtained from the U.S. National Archives and various secondary sources. The dissertation concludes that an understanding of American artillery observation is critical to a more complete comprehension of the battle of Okinawa. These mid-ranking (and largely middle class) soldiers proved capable of adjusting to the demands of combat conditions. They provide a unique and understudied perspective of the entire battle.

Seabourn Cruise Line: A Case Study in Achieving Quality

Just about everyone who ranks cruise lines puts Seabourn first on the list. The readers of Conde Nast Traveler ranked it the world's top cruise line for three consecutive years and fifth in their survey of the top 100 overall travel experiences. Of special interest to hospitality professionals is Seabourn's 98.5 percent score for service- higher than any other vacation experience in the world.

Off-line and on-line affective recognition of a computer user through a biosignal processing approach

Physiological signals, which are controlled by the autonomic nervous system (ANS), could be used to detect the affective state of computer users and therefore find applications in medicine and engineering. The Pupil Diameter (PD) seems to provide a strong indication of the affective state, as found by previous research, but it has not been investigated fully yet. ^ In this study, new approaches based on monitoring and processing the PD signal for off-line and on-line affective assessment ("relaxation" vs. "stress") are proposed. Wavelet denoising and Kalman filtering methods are first used to remove abrupt changes in the raw Pupil Diameter (PD) signal. Then three features (PDmean, PDmax and PDWalsh) are extracted from the preprocessed PD signal for the affective state classification. In order to select more relevant and reliable physiological data for further analysis, two types of data selection methods are applied, which are based on the paired t-test and subject self-evaluation, respectively. In addition, five different kinds of the classifiers are implemented on the selected data, which achieve average accuracies up to 86.43% and 87.20%, respectively. Finally, the receiver operating characteristic (ROC) curve is utilized to investigate the discriminating potential of each individual feature by evaluation of the area under the ROC curve, which reaches values above 0.90. ^ For the on-line affective assessment, a hard threshold is implemented first in order to remove the eye blinks from the PD signal and then a moving average window is utilized to obtain the representative value PDr for every one-second time interval of PD. There are three main steps for the on-line affective assessment algorithm, which are preparation, feature-based decision voting and affective determination. The final results show that the accuracies are 72.30% and 73.55% for the data subsets, which were respectively chosen using two types of data selection methods (paired t-test and subject self-evaluation). ^ In order to further analyze the efficiency of affective recognition through the PD signal, the Galvanic Skin Response (GSR) was also monitored and processed. The highest affective assessment classification rate obtained from GSR processing is only 63.57% (based on the off-line processing algorithm). The overall results confirm that the PD signal should be considered as one of the most powerful physiological signals to involve in future automated real-time affective recognition systems, especially for detecting the "relaxation" vs. "stress" states.^

On-Line Real-Time Service-Oriented Task Scheduling Using TUF

We present our approach to real-time service-oriented scheduling problems with the objective of maximizing the total system utility. Different from the traditional utility accrual scheduling problems that each task is associated with only a single time utility function (TUF), we associate two different TUFs—a profit TUF and a penalty TUF—with each task, to model the real-time services that not only need to reward the early completions but also need to penalize the abortions or deadline misses. The scheduling heuristics we proposed in this paper judiciously accept, schedule, and abort real-time services when necessary to maximize the accrued utility. Our extensive experimental results show that our proposed algorithms can significantly outperform the traditional scheduling algorithms such as the Earliest Deadline First (EDF), the traditional utility accrual (UA) scheduling algorithms, and an earlier scheduling approach based on a similar model.

Characterization and modeling of multi-conductor transmission line using Finite-difference Time-Domain method

A two-dimensional, 2D, finite-difference time-domain (FDTD) method is used to analyze two different models of multi-conductor transmission lines (MTL). The first model is a two-conductor MTL and the second is a threeconductor MTL. Apart from the MTL's, a three-dimensional, 3D, FDTD method is used to analyze a three-patch microstrip parasitic array. While the MTL analysis is entirely in time-domain, the microstrip parasitic array is a study of scattering parameter Sn in the frequency-domain. The results clearly indicate that FDTD is an efficient and accurate tool to model and analyze multiconductor transmission line as well as microstrip antennas and arrays.