The emergent resistance of bacteria against the conventional antibiotics has motivated the search for novel antimicrobial agents. Nature abounds with a number of antimicrobial peptides that are a part of our innate immune system and protect us against a variety of pathogenic bacteria. While they are broad-spectrum in their activity and show less drug-resistance induction, their intrinsic metabolic stability limits their potential therapeutic applications. Herein we describe the development of novel broad-spectrum bioactive antimicrobial peptidomimetics AA-peptides. AApeptides were designed based on chiral PNA backbone. Substitution of nucleobases yields AApeptides that are resistant to proteolysis and capable of mimicking peptides. Two types of AApeptides are discussed in this dissertation "[aacute]-AApeptides" and " ã-AApeptides" The therapeutic potential of these AApeptides were accessed by conducting antibacterial assays against a series of both gram-positive, gram-negative bacteria and fungi. These oligomers were characterized using MALDI-TOF and Circular Dichroism spectroscopy (CD). Their invitro toxicity was evaluated against human erythrocytes .We attempted to study their mechanism of action via membrane depolarization assay. We have successfully identified them as antimicrobial agents, pro-inflammatory immune response suppressing agents and as anti-biofilm agents.

Alors que la base des interactions hôte-pathogène chez les plantes terrestres sont désormais très bien décrites, que ce soit au niveau physiologique, métabolique ou moléculaire, chez les macroalgues marines les investigations n'en sont qu'à leur début. Chez l'algue brune L. digitata, les études sur les réponses de défense suite à l'élicitation par les oligoguluronates, ont permis d'appréhender la transduction du signal conduisant à la production d'un burst oxydant, ainsi que la mise en place tardive de la résistance. Le métabolisme halogéné est particulièrement actif chez cette algue. Il semble être impliqué dans les réponses de défense comme en témoigne l'émission d'iode moléculaire et de composés organiques halogénés volatils, suite à l'élicitation par les oligoguluronates. Dans ce contexte, j'ai étudié la régulation du transcriptome de L. digitata en réponse à l'élicitation par les oligoguluronates, par hybridation soustractive suppressive, macroarray et PCR Quantitative. J'ai ainsi identifié dix gènes codant notamment des haloperoxydases, des thiorédoxines et des enzymes de la voie des pentoses phosphates. L'induction de ces gènes suite à l'élicitation suggère une gestion originale du stress oxydant chez L. digitata. L'implication du métabolisme halogéné dans les réponses de défense a été mise en avant par l'expression différentielle de certains gènes codant des haloperoxydases. La régulation spécifique de certains de ces gènes apporte la première preuve expérimentale que les différents membres d'haloperoxydases ont évolué vers des fonctions biologiques spécifiques. L'identification de ces gènes comme marqueurs moléculaires des réponses de défense a permis d'établir la première modélisation de la signalisation intracellulaire conduisant à l'activation de gènes de défense de L. digitata et de souligner le rôle du peroxyde d'hydrogène dans plusieurs voies distinctes. Les outils développés et les marqueurs moléculaires identifiés permettront d'approfondir la compréhension des réponses de défense des macroalgues dans le contexte des interactions biotiques et de la signalisation à distance.

Apologia is a rhetorical genre of self-defense. As such, apologetic discourse involves a response to an attack upon an individual's character or worth as a human being. Los Angeles Police Detective Mark Fuhrman encountered such a attack during the 0.J. Simpson murder trial. The purpose of this analysis is to determine whether the rhetorical strategies indicative of apologia are used to perform masculine facework in Mark Fuhrman's Murder In Brentwood. By means of critical analysis, apologia can be established as a masculine face-saving strategy. The analysis takes place in two stages. First, the connection between facework and apologia is established. The four strategies of Hodgins, Liebeskind, and Schwartz (1996), Sandra Petronio's examples of defensive strategies, and three strategies in William L. Benoit's (1995) typology of image restoration strategies provide the first two units of analyses. Second, questions gleaned from the research of Noreen Kruse (1977) determined the topics to which Fuhrman devotes the most time in his discourse and the motivational drives that shape that discourse. By integrating the findings of both analyses, the masculine face Fuhrman endeavors to save emerges. / Graduation date: 1999

Thesis (LL. M.)--Judge Advocate General's School, United States Army, 1995. / "April 1995." Typescript. Includes bibliographical references. Also issued in microfiche.

Takeovers are used as a mean for companies to grow and gain entry to new markets. Hos-tile takeovers, apart from the friendly takeovers, is when an acquirer tries to takeover a corporation against the will of management, shareholders and board of directors of the target company. All listed companies run the risk of being a target for a hostile takeover, and to-day many companies are trying to involve actions protecting them from possible threats. Lacking protection can evolve into a costly defense program with actions with fictitious names such as White knight, Pac-man and Poison pills. The purpose with this thesis is to describe and analyze hostile takeovers and hostile takeover attempts in Sweden, and the defense tactics involved in the process. In this thesis a deductive approach will be used, where theories are used as guidance when searching for explanations. On the basis of the defense tactics described in the theory empirical data has been gathered with the purpose of seeing if the expectations reflect reality, and conclusions about their efficiency. The thesis has used a quantitative research method where the focus is on what, where and when. The aim of the thesis is to classify targeted features and count them, with the intent to construct statistical models with an underlying purpose of explaining what is observed. The most frequently used defense tactic used by the target company in a hostile takeover in the Swedish market is to attack the logic of the bid. Around 56 percent of the targets in a hostile takeover or a hostile takeover attempt have used this defense tactic to avoid a hos-tile takeover. The aim with an implemented defense tactic is to avoid a hostile bid or making it impossible for the hostile bidder to proceed with the bid and close the deal. The White knight and White squire defense tactic is the most successful defense tactic when it comes to not finalizing the hostile bid, in 90 percent of the cases the deal is not closed if the target chooses to use a White knight as a defense tactic. A secondary objective with a defense tactic is to force the hostile bidder to increase the bid and pay more for the target company. In the Swedish market, the use of Corporate restructuring as a defense tactic has made the hostile bidder to increase the bid in 67 percent of the cases and the use of Positive public information has forced the hostile bidder to increase the bid in more than half of the cases. The question why these defense tactics are the most frequently used strategies, is explained by two variables. The first one is the cost and simplicity variable, where Attack the logic of the bid and Public information ends up. These defense tactics are cost efficient and can be seen as natural step for the target company when deciding not to approve of the offer made by the acquirer. The second variable is the proven efficiency, where the target knows if implementing this defense tactic the risk of being acquired by the hostile bidder is relatively low, a good example of this is the White knight defense.

The handwriting was on the wall when I arrived at the Defense Information School in December, 1977. I had been brought in after receiving a masters in management to redesign the public affairs supervisors course and the public affairs officer course stressing management rather than a skill orientation. During that process, which lasted two years, I became increasingly involved with the problem addressed in this creative project. The problem plagued Army schools throughout the Training and Doctrine Command.Under the ISD program, initially adopted in 1975, Army educators were forced to ask themselves three questions: What should be the role of the instructor in improving instruction; is there a better way to plan for effective instruction; and can education really be improved, within the limitations of available funds, personnel, and facilities?Working now as the organizational effectiveness consultant at DINFOS, I turned those questions around into positive statements which goal-directed educational administrators could deal with. Essentially I proposed that successful innovation in education requires at least three elements:(1) instructors who are deeply concerned about their teaching effectiveness and who are motivated by a desire for improvement, (2) administrators who willingly encourage and support those instructors, and (3) a carefully designed plan for developing improved instructional practices. Of these three elements, the greatest shortcoming at DINFOS was found in the third.Student evaluations and the North Central accrediting team underscored our instructors' concern. The school administration has demonstrated its willingness to innovate. TRADOC had provided the umbrella plan-ISD. The problem was instructor interface with the ISD process through our Directorate of Training Developments which is tasked to bring ISD to fruition at DIINIFOS.Many suggestions were tried. For example, this seemed to be a textbook case for Management by Objectives (MBO). It did not work because instructor time is at a premium. Finally, my recommendation was adopted and seems headed for success. This remedy is essentially two-fold. It makes, as a matter of policy, ISD the operant mode for all instruction and is outlined in the Operations Manual as such. Secondly, all newly-assigned instructors and other instructors without benefit of this training, must attend a class in ISD where they must demonstrate knowledge of the process and, in fact, write behavioral objectives before being certified to instruct at the school.

A new paradigm for the treatment of infectious disease is through the modulation of innate immune responses. In this capacity, host defense peptides (HDPs) and synthetic Toll-like receptor 9 (TLR9) ligands have the greatest demonstrated potentials. The work presented here considers mechanisms for the improvement of these treatments through optimization, or in the case of HDPs the minimization, of the interactions of these ligands with sensory receptors.<p>Toll-like Receptor 9 activates the innate immune system in response to microbial DNA or immune-modulating oligodeoxynucleotides. While cell stimulation experiments demonstrate the preferential activating ability of CpG-containing nucleic acids, direct binding investigations have reached contradictory conclusions regarding the sequence-specificity of TLR9 ligand binding. To address this discrepancy the characterization of human TLR9 ligand binding properties is reported. TLR9 has a high degree of ligand specificity in being able to discriminate not only CpG dinucleotides, but also higher order six nucleotide motifs that mediate species-specific activation. However, TLR9 ligand binding is also functionally influenced by nucleic acids in a sequence-independent manner both in vitro and in cell proliferation experiments. A model is proposed in which TLR9 activation is mediated specifically by CpG-containing ligands while sensitivity of the receptor is modulated by the absolute concentration of nucleic acids in a sequence-independent fashion. <p>Host defense peptides are among the leading candidates to combat antibiotic resistant bacterial strains. Recently, HDPs have been demonstrated to function as ligands for the bacterial sensory kinase PhoQ resulting in the induction of virulence and adaptive responses. Thus, concerns have been raised regarding therapeutic applications of HDPs. Here a methodology is described that permits discrimination and quantification of the distinct, but related, peptide behaviors of direct antimicrobial activity and PhoQ ligand potential. Utilizing peptide derivatives of the model HDP Bac2A it is demonstrated that antimicrobial efficiency is significantly, and inversely, related to PhoQ ligand efficacy. This provides a rational basis for HDP selection with greater therapeutic potential and minimized potential for initiation of bacterial resistance.

mPIP is a mouse homologue of human PIP/GCDFP-15 which is an established marker of both malignant and benign pathological conditions of the mammary gland. mPIP gene expression has been identified in both lacrimal and salivary glands of healthy mice and the mPIP protein has been detected in saliva. The mPIP protein has been found to bind oral bacteria, showing the highest affinity for streptococci, suggesting a potential function of mPIP in the non-immune host defense in the mouse oral cavity. Since the exact functions of mPIP are still unknown, we examined the roles of mPIP through both in vitro and in vivo studies, specifically to address the possible role of this protein in non-immune host response through modulating the oral flora. The in vitro studies were primarily focused on elucidation of the consequences of interaction between mPIP and oral bacteria, in particular to examine whether mPIP plays a role in bacterial aggregation. The in vivo studies addressed the roles of mPIP through the analysis of an mPIP knockout mouse model generated in our laboratory. Following confirmation of the null mutation, the delineating the phenotype of this model was pursued through morphopathological analysis as well as examination of the impact of the lack of mPIP on the mouse oral flora. The null mutation in the mPIP knockout mice was confirmed by both the gene and protein analysis. Histological analysis revealed lymphocytic proliferation in both the submaxillary and prostate glands of the mPIP knockout mice. In addition, both quantitative and composition differences in the oral flora of mPIP knockout mice were identified when compared with wild-type controls. Specifically, a higher proportion of the oral bacteria of mPIP knockout mice were found to belong to genus Streptococcus and certain genera were found to be absent from the oral cavity of these mice. The effect of knockout mouse saliva, which lacks mPIP, on the aggregation of oral bacteria was compared to wild-type mouse saliva. Our data suggests that mPIP contributes to saliva-induced bacterial aggregation. While oral flora has multiple functions, including protection against infection, mPIP might play a role in the non-innate host defense through modulating the resident oral flora in the mouse. The identification of lymphocytic proliferation in submaxillary and prostate glands of mPIP knockout mice suggests that mPIP might also interfere with lymphocyte activity, playing a possible immunomodulatory role. / May 2008

The emergence of pathogens resistant to available drug therapies is a pressing global health problem. Antimicrobial peptides (AMPs) may potentially form new therapeutics to counter these pathogens. AMPs are key components in the mammalian innate immune system and are responsible for both direct killing and immunomodulatory effects in host defense against pathogenic organisms. This thesis describes computational methods for the identification of novel natural and synthetic AMPs. A bioinformatic resource was constructed for classification and discovery of gene- coded AMPs, consisting of a database of clustered known AMPs and a set of hidden Markov models (HMMs). One set of 146 clusters was based on the mature peptide sequence, and one set of 40 clusters was based on propeptide sequence. The bovine genome was analyzed using the AMPer resources, and 27 of the 34 known bovine AMPs were identified with high confidence and up to 69 AMPs were predicted to be novel peptides. One novel cathelicidin AMP was experimentally verified as up-regulated in response to infection in bovine intestinal tissue. A chemoinformatic analysis was performed to model the antibacterial activity of short synthetic peptides. Using high-throughput screening data for the activities of over 1400 peptides of diverse sequence, quantitative structure-activity relation (QSAR) models were created using artificial neural networks and physical characteristics of the peptide that included three-dimensional atomic structure. The models were used to predict the activity of a set of approximately 100,000 peptide sequence variants. After ranking the predicted activity, the models were shown to be very accurate. When 200 peptides were synthesized and screened using four levels of expected activity, 94% of the top 50 peptides expected to have the highest level of activity were found to be highly active. Several promising candidates were synthesized with high quality and tested against several multi- antibiotic-resistant pathogens including clinical strains of Pseudomonas aeruginosa, Staphylococcus aureus, Enterococcus faecalis and Escherichia coli. These peptides were found to be highly active against these pathogens as determined by minimal inhibitory concentration; this serves as independent confirmation of the effectiveness of high-throughput screening and in silico analysis for identifying peptide antibiotic drug leads.

A new paradigm for the treatment of infectious disease is through the modulation of innate immune responses. In this capacity, host defense peptides (HDPs) and synthetic Toll-like receptor 9 (TLR9) ligands have the greatest demonstrated potentials. The work presented here considers mechanisms for the improvement of these treatments through optimization, or in the case of HDPs the minimization, of the interactions of these ligands with sensory receptors.<p>Toll-like Receptor 9 activates the innate immune system in response to microbial DNA or immune-modulating oligodeoxynucleotides. While cell stimulation experiments demonstrate the preferential activating ability of CpG-containing nucleic acids, direct binding investigations have reached contradictory conclusions regarding the sequence-specificity of TLR9 ligand binding. To address this discrepancy the characterization of human TLR9 ligand binding properties is reported. TLR9 has a high degree of ligand specificity in being able to discriminate not only CpG dinucleotides, but also higher order six nucleotide motifs that mediate species-specific activation. However, TLR9 ligand binding is also functionally influenced by nucleic acids in a sequence-independent manner both in vitro and in cell proliferation experiments. A model is proposed in which TLR9 activation is mediated specifically by CpG-containing ligands while sensitivity of the receptor is modulated by the absolute concentration of nucleic acids in a sequence-independent fashion. <p>Host defense peptides are among the leading candidates to combat antibiotic resistant bacterial strains. Recently, HDPs have been demonstrated to function as ligands for the bacterial sensory kinase PhoQ resulting in the induction of virulence and adaptive responses. Thus, concerns have been raised regarding therapeutic applications of HDPs. Here a methodology is described that permits discrimination and quantification of the distinct, but related, peptide behaviors of direct antimicrobial activity and PhoQ ligand potential. Utilizing peptide derivatives of the model HDP Bac2A it is demonstrated that antimicrobial efficiency is significantly, and inversely, related to PhoQ ligand efficacy. This provides a rational basis for HDP selection with greater therapeutic potential and minimized potential for initiation of bacterial resistance.