Yes

The type VII secretion system is a protein export pathway linked to diverse phenotypes in both Actinobacteria and Firmicutes. The Actinobacterial subtype of the T7SS, referred to as T7SSa, has been shown to play a critical role in various aspects of Mycobacterial life including virulence, conjugation, and metal homeostasis. The T7SSb of Firmicutes bacteria on the other hand has similarly been shown to influence virulence but by the direct growth inhibition of competitor bacteria. Structure-function analyses of the T7SSa apparatus as well as various effectors and chaperones have begun to build a more mechanistic understanding of how T7SSa functions. In contrast, we know little of how the T7SSb functions despite its noted importance to both pathogens and environmental bacteria such as Bacillus, Staphylococcus, Enterococcus, and Streptococcus. During my thesis work, I have addressed several gaps in our understanding of T7SSb function. The three major questions that I have studied are: (1) how do T7SSb immunity proteins inhibit the toxicity of their cognate toxins, (2) how does the T7SSb export effectors through the thick Gram-positive cell wall, and (3) what is the role of chaperone proteins in facilitating T7SSb effector export? / Thesis / Doctor of Philosophy (PhD) / Bacteria require space and various nutrients to survive and grow and must therefore compete against other bacteria for access to these resources. To gain advantage over their competitors, many bacteria have developed molecular weapons that target and kill other closely related bacteria. Some of these weapons take the form of protein secretion machines that export antibacterial toxins. Gram-positive bacteria use the type VIIb secretion system (T7SSb) to inhibit the growth of other Gram-positive bacteria. In this work, I explore several aspects of T7SSb including: (1) how toxins are inhibited by immunity proteins, (2) how toxins are secreted through the cell envelope, and (3) how toxins are recognized by the secretion apparatus. The goal of this work is to better understand how T7SSb functions at the molecular level.

The purpose of this study was to trace the development of case law regarding sexual harassment in elementary and secondary public schools through a survey of state, circuit, and federal court districts, and United States Supreme Court decisions. To trace these laws, the study examined nationwide sexual harassment court decisions from 1988 to present. Sexual harassment is a critical problem in the school environment. Recent surveys indicate that sexual harassment has reached epidemic proportions in our schools and classrooms. Sexual harassment occurs daily on buses, in hallways, gymnasiums, classrooms, and on the playgrounds. Oftentimes, sexual harassment goes unreported by students. More importantly, when sexual harassment is reported, the school does virtually nothing to assure studetns that their complaints will be taken seriously. Sexual harassment can contribute to serious harm to the victim, their families, other classmates, the school, and the school district. A review of related literature reveals that students who are sexually harassed suffer from behavioral, emotional, academic, and physical consequences. Without the support and intervention of school officials, students may not be able to overcome the impact of sexual harassment. Several therories and models were reviewed in the study to examine possible explanations for incidence of sexual harassment in public schools. It should be noted that no single theory or model offered a precise explanation of the reasons students are harassed in schools. Several landmark and key court cases regarding sexual harassment in public school were reviewed. Also, key federal legislative statutes and guidelines that provide the framework for judicial decisions on sexual harassment in public schools were reviewed. It was determined that four U. S. Supreme Court cases set precedence for lower courts to follow in adjudicating decisions of sexual harassment in public schools. The judicial decrees protect students in the school environment and are specific about the roles and responsiblities of the school district, administrators, and teachers. Additionally, the courts have taken important and significant steps towards making school districts officials liable for not effectively addressing sexual harassment in the schools. Based on the broad scope of the study and the conclusions, several educational, prevention, and intervention strategies were recommended for students, parents, teachers, administrators, school districts, the community, and stakeholders. If these recommendations are implemented in school districts nationwide, the incidence and prevalence of sexual harassment in public school should decrease significantly.

La mucopolysaccharidose de type VII (MPS VII) est une maladie génétique de surcharge lysosomale provoquée par une déficience en β-D-glucuronidase (β-glu). β-glu est impliquée dans la cascade de dégradation et de recyclage des glycosaminoglycannes (GAGs). Sa déficience provoque une accumulation vésiculaire de GAGs non dégradés engendrant in fine la mort cellulaire. Notre but est de développer et de tester la pertinence des vecteurs adénoviraux canins helper-dépendant (HD-CAV-2) pour traiter la neurodégénération provoquée par la MPS VII dans le cadre d'une thérapie génique dans le SNC de chien. Parce que les vecteurs CAV-2 transduisent préférentiellement les neurones et qu'ils sont transportés de manière rétrograde le long des axones, leurs distributions dans tout le SNC permettraient de délivrer largement la β-glu dans tout le parenchyme cérébral. Nous avons alors étudié la sureté, la durée d'expression, l'efficacité et la possible réversion du phénotype après injections dans le SNC de chiens MPS VII d'un HD-CAV-2 exprimant le gène humain de la GUSB : HD-RIGIE. Des études préliminaires ont montré la faisabilité du transfert des vecteurs HD-CAV-2 dans le SNC, qu'ils induisaient une réponse immunitaire minimale et qu'ils transduisaient préférentiellement, efficacement et largement les neurones.Nous avons produit un HD-RIGIE de qualité pour les injections intracérébrales et nous avons analysé son efficacité sur l'accumulation de GAGs non dégradés. Les injections de HD-RIGIE montrent une augmentation générale de l'activité β-glu dans tout le SNC des chiens MPS VII (sites d'injections et structures éloignées comme le cortex) et ce 1 mois ou 4 mois après les injections. L'analyse de la GFP confirme une distribution globale de HD-RIGIE dans le SNC d'animaux de grande taille. De plus, grâce aux propriétés intrinsèques de la β-glu (transport rétrograde et phénomène de libération/recapture), nous avons observé une diminution générale de l'accumulation vésiculaire neuronale des GAGs non dégradés dans tout le parenchyme cérébral. D'autre part grâce à la stratégie d'isolement et de non vaccination des chiens MPS VII, nous n'observons ni de réponse immunitaire humorale, ni d'aggravation de l'inflammation du parenchyme. / Mucopolysaccharidosis type VII (MPS VII) is a rare inherited lysosomal storage disease caused by β-D-glucuronidase (β-glu) deficiency. β-glu is involved in the physiological turnover of glycosaminoglycans (GAGs). Its deficiency causes accumulation of undegraded GAGs inside vesicles leading to cell death. Our goals are to develop and test the clinical relevance of helper-dependant (HD) canine adenovirus type 2 (CAV-2) vectors to treat neural degeneration caused by MPS VII in a dog model. Because CAV-2 targets preferentially neurons and traffics via axons, the distribution of the transgene throughout the CNS will allow widespread delivery of the missing lysosomal enzyme in these disorders with a minimum number of injections. We tested a HD-CAV-2 vector expressing the human GUS gene in the canine model of MPS VII for their safety, efficacy, duration of expression and possible reversion of the MPS VII induced symptoms.A previous study based on HD-CAV-EGFP vector injections in MPS VII-/- and healthy dogs showed that we are now able to inject HD-CAV-2 in the dog brain, have a minimal induction of the immune response, an efficient transduction of the neurons and an efficient biodistribution of transduced cells. After the production of a suitable vector (HD-RIGIE) for injections in the CNS of MPS VII dogs we analysed its efficiency on GAGS storage in neurons.Injections of HD-RIGIE showed after 1 month or 4 months post injections a widespread increase in general level of β-glu activity, in the sites of injections and in distant areas such as cortex. Analysis of GFP, also permit to observe a widespread biodistribution of the vector. Because of β-glu property of cross-correction we observed a global decreased in GAGs storage in the entire MPS VII brains. Finally, the dogs did not present humoral immune response and no aggravation of inflammation

Ve své diplomové práci se jsem se pokusil popsat politickou situaci v Itálii v první třetině 14. století se zaměřením na působení významných osobností Lucemburského rodu v této oblasti. Pro zpracování jsem zvolil léta 1308-1313, 1330-1333 a rok 1337. Zaměřil jsem se hlavně na osobu Jindřicha IV. Lucemburského, jeho nástup k moci, volbu říšským králem a následnou cestu za císařskou korunou. V práci rozebírám reformy, které Jindřich použil při správě Italských měst během cesty na Řím. Dále se zaměřuji na odezvu, kterou jeho působení vyvolávalo u příslušníků dvou hlavních politických směrů v Itálii, tedy u guelfů a ghibelinů, i u francouzského krále Filipa IV. a u papeže Klimenta V. pobývajícího tou dobou v Avignonu. Dále se zaobírám pokusem českého krále Jana Lucemburského a jeho syna Karla o založení Lucemburské signorie na území Lombardie, Toskánska a Emilia Romagna a jejich neúspěšné snahy o udržení tohoto území pod Lucemburskou nadvládou. A Karlovo krátkodobým pokusem o získání území v severní Itálii v roce 1337. Opět se věnuji vlivu jejich působení na politiku měst zmíněných oblastí. Také se okrajově zaměřuji na vliv Jindřicha Lucemburského na spisovatele Danta Alighieriho a jeho dílo "De Monarchia" s Jindřichovo tažením do Říma jako předlohou pro vznik tohoto díla.

The United Nations Security Council is primarily responsible for maintenance of international peace and security according to the Charter of the United Nations. To achieve this goal, it is authorized to adopt resolutions binding on member states under Chapter VII of the Charter. Considering the character of situations to which the Security council responds through its Chapter VII resolutions, this legal instrument often stirs emotions among the general public. But what does the law itself say about the Chapter VII resolutions? What is their legal nature and value in the field of public international law? This thesis provides an analysis of the Chapter VII resolutions, offers a definition of a Chapter VII resolution and analyzes some specific resolutions by which the Security Council acted more as a quasi-judicial or a quasi-legislative body. Through analysis of content limits of the Security Council powers, the thesis endeavors to confirm that binding nature of the Chapter VII resolutions as well as an obligation of member states to give effect to those resolutions do have its boundaries; and despite the special role of the Security Council in the field of public international law, its Chapter VII resolutions cannot have unlimited content. First of all, the resolutions cannot be in conflict with...

[Truncated abstract] Intracellular communication between osteoclasts and osteoblasts is imperative to maintain bone integrity. A myriad of molecules are responsible for regulating osteoblast and osteoclast activity. In particular, it is well documented that osteoblast-derived factors are crucial in directly controlling osteoclast formation and function. Since bone formation is coupled to bone resorption, it would be expected that osteoclasts also have some role in regulating the growth and function of osteoblast cells. However, despite extensive research upon osteoclast and osteoblast biology, the mechanisms by which osteoclasts regulate osteoblast growth and function is not well understood. In an attempt to further elucidate the mechanisms by which osteoclasts and osteoblasts communicate, the technique of subtractive hybridisation was used to identify a novel osteoclastderived factor identical to that of mouse Seminal Vesicle Secretion VII (SVS VII). Previous characterisation of the gene in bone demonstrated that SVS VII was abundantly and specifically expressed by mature osteoclasts (Phan, 2004). Additional research hinted that SVS VII acted as a novel osteoclast-derived factor, that by paracrine mechanisms, targeted osteoblast function (Phan, 2004). However, it remained open as to whether the SVS VII molecule did uniquely target the osteoblast, and whether this interaction influenced bone formation in vivo. Therefore, this thesis endeavoured to functionally characterise the role of the SVS VII molecule in the bone environment. ... Further work is needed to identigy a clear consensus binding sequence, to determine the specificity of the interaction between SVS VII protein and each phage clone, and to isolate a specific binding partner for SVS VII. In conclusion, the studies of this thesis sought to characterise the significance of SVS VII expression by mature osteoclasts, relative to its effects on osteoblast behaviour, but failed to conclusively determine a role for SVS VII in bone. Given that the effects of SVS VII on in vitro osteoblast activity and function are minimal, it is doubtful that SVS VII primarily acts as a paracrine factor integral to osteoblast function. Therefore, these findings conflict with those presented previously (Phan, 2004). However, it was demonstrated that SVS VII treatment was associated with in vivo effect on the skeleton, suggesting that SVS VII may target other elements of the bone microenvironment. Via mechanisms not yet understood, which possibly involves additional factors of the bone 11 extracellular matrix, SVS VII may target a subset of osteoprogenitor cells within the bone environment and act to regulate their proliferation. Therefore, SVS VII may enhance osteogenic precursor cell number at sites of bone formation which would increase the pool of cells that can differentiate down the osteoblast linage and contribute to bone formation. In this regard, SVS VII might function in a manner homologous to the Ly-6 molecule Sca-1 and act as an important factor that maintains a balance between the bone formation and resorption process. Clearly, more work focusing on alternative facets of bone biology is needed to identify whether there is a significant role for SVS VII in skeletal tissue.

Mycobacterium abscessus est une mycobactérie à croissance rapide, et un pathogène opportuniste responsable d’infections pulmonaires notamment chez les patients atteints de la mucoviscidose, et d’infections cutanéomuqueuses. La source de contamination pourrait être environnementale mais des contaminations interhumaines ne sont pas exclues. Les amibes environnementales pourraient jouer un rôle de réservoir. M. abscessus est capable de résister aux mécanismes de défense bactéricides des phagocytes environnementaux et humains. Le génome complet de M. abscessus a été séquencé mettant en évidence de nombreux facteurs de virulence non mycobactériens. Certains sont des facteurs de virulence connus dans le monde bactérien, comme la phospholipase C ou le facteur de captation du magnésium MgtC. Ces facteurs ont été montré induits en présence d’amibes, mais ne peuvent à eux seuls expliquer la survie intracellulaire et la virulence de M. abscessus. Nous avons donc, au cours de ce projet, criblé une banque de mutants générée par transposition chez M. abscessus, à la recherche de mutants dénués de croissance intracellulaire en amibes et macrophages. Cette approche a permis d’identifier, de façon majeure, 5 gènes du locus ESX-4 de M. abscessus codant un système de sécrétion de type VII avec tous ces composants cœur conservés. Pour mieux comprendre la contribution d’ESX-4 dans la survie intracellulaire de M. abscessus, un mutant obtenu par double recombinaison au sein du gène eccB4 dans la souche type de M. abscessus (ΔeccB4) a été construit. EccB4 est un élément structurel central du système de sécrétion codé par ESX-4. ΔeccB4 présente un défaut de survie au sein des cellules, lié à déficit de blocage de l’acidification phagosomale ainsi qu’un défaut de dégradation de la membrane phagosomale, empêchant un contact phagosome-cytosol. Ce travail a permis de révéler pour la première fois dans le monde mycobactérien le rôle d’un locus ancestral de sécrétion ESX-4 au sein d’une mycobactérie. L’étude des protéines secrétées par ce locus est actuellement en cours au laboratoire, afin d’envisager des approches thérapeutiques et vaccinales pour contrer cette mycobactérie multirésistante aux antibiotiques. / Mycobacterium abscessus is a fast growing mycobacterium, and an opportunistic pathogen responsible for lung infections particularly in patients with cystic fibrosis, and for mucocutaneous infections. The source of contamination could be environmental but human-to-human contaminations are not excluded. Environmental amoeba could play a role as a reservoir. M. abscessus is able to resist to the bactericidal defense mechanisms of environmental and human phagocytes. The complete genome of M. abscessus has been sequenced and presents numerous non-mycobacterial virulence factors. Some are known virulence factors in the bacterial world, such as phospholipase C or the magnesium uptake factor MgtC. These factors have been shown to be induced in the presence of amoeba, but cannot alone explain the intracellular survival and virulence of M. abscessus. We thus, in the course of this project, screened a library of mutants generated by transposition in M. abscessus, in search of mutants lacking intracellular growth in amoeba and macrophages. This approach made it possible to identify, in a major way, 5 genes of the ESX-4 locus of M. abscessus encoding a type VII secretion system with all these conserved core components. In order to better understand the contribution of ESX-4 to the intracellular survival of M. abscessus, a mutant obtained by double recombination within the eccB4 gene in the M. abscessus type strain (ΔeccB4) was constructed. EccB4 is a central structural element of the secretion system encoded by ESX-4. ΔeccB4 has a defect of survival within the cells, linked to deficiency of blockage of the phagosomal acidification as well as a defect of degradation of the phagosomal membrane, preventing phagosome-cytosol contact. This work made it possible to reveal for the first time in the mycobacterial world the role of an ancestral locus ESX-4 secretion within a mycobacterium. The study of the proteins secreted by this locus is currently underway in the laboratory, in order to consider therapeutic and vaccine approaches to counter this multiresistant antibiotic mycobacterium.

Thesis (DTh)--Stellenbosch University, 2001 / ENGLISH ABSTRACT: This study aims to interpret Romans 7 with special reference to the law. Both Romans 7 and the law in Paul are very difficult to understand. However, both are important for an understanding of Pauline theology and the gospel. In the past historical critical analyses were usually done in order to solve problematic passages like Romans 7 in Paul's letters. In this study a socio-rhetorical analysis is utilized. To start with, previous research is briefly dealt with in order obtain an overall picture of the understanding of the law in the past. From this overview more than ten problem areas are identified. Then, socio-rhetorical analysis is briefly explained. This is a multi-dimensional and multi-disciplinary method developed by Vernon Robbins, which sees the text as having various textures. In this research the method is utilized with some modifications. In chapter 4 the macrostructure of Romans is established by means of epistolary analysis and rhetorical analysis. Next, Rom. 7 is established as a rhetorical unit within Rom. 5-8 as the broader co-text of Rom. 7. After that the rhetorical situation of Romans is discussed, as well as some of Paul's rhetorical devices and styles. Finally, the rhetorical species of Romans is determined as deliberative rhetoric. From chapters 5 to 7, Rom. 7 is analyzed, using different textual analyses. In an analysis of inner texture repetitive-progressive texture, opening-middle-closing texture, and argumentative texture are discussed. Here an enthymemic analysis is used in order to chart Paul's argumentative flow of thought. From this it is concluded that Rom. 7: 1-6 is an analogy, which is an important tool for argumentation, and that the present tense in Rom. 7:14-25 functions as part of a combination of autobiographical-typical-rhetorical features for the purpose of argumentation. In the analysis of intertexture the scriptural intertexture is investigated: recitation with omission and thematic elaboration. In the cultural intertextual analysis some Jewish cultural intertextures are noted, namely, Rom. 7:8-10 as an allusion to both Gen. 3 and Exod. 3, Paul's usage of the "1," the law, slavery image, and the evil inclination. It is also interesting that Rom. 7: 15 & 19 and the "I" are allusions to Greek tragedy, sin as power, and slavery as Greco-Roman cultural intertexture. In analyzing the social intertexture it can be concluded that the marriage analogy is closer to Jewish marriage than to Greco-Roman marriage. The final analysis is an investigation of the theological texture. Here salvation history and the covenant of God are first dealt with in order to get to grips with Paul's theological world. Then, Pauline hamartiology, anthropology, and finally, nomism are investigated. The conclusion is that VOl-lOS' in Rom. 7 mostly denotes the universal moral law of God, both written and unwritten, not just the Mosaic law; though in some cases it denotes "principle" or "rule" as in vv. 21-25. Rom. 7 as a whole is a refutation of the objection or misunderstanding that might be raised regarding Paul's statements of the law in previous chapters. In Rom. 7 Paul elaborates the relationship between believers and the law, and the function of the law in relation to sin in an unregenerate person. In so doing, he vehemently denies that the law is sin, and vividly indicates the function of the law using his own experience. / AFRIKAANSE OPSOMMING: Hierdie navorsing fokus op Rom. 7 met spesifieke verwysing na die wet. Hoewel sowel die wet by Paulus en Rom. 7 moeilike onderwerpe is om te verstaan, is beide van besondere belang vir In verstaan van die teologie van Paulus en die evangelie. In die verlede is gewoonlik van histories-kritiese studies gebruik gemaak in die bestudering van sulke probleme. Hier word van In sosio-retoriese benadering gebruik gemaak. Daar word eers aandag gegee aan vorige navorsing om In beeld te kry van die wyse waarop die wet in die verlede verstaan is. Na aanleiding van hierdie oorsig word meer as tien probleemareas geidentifiseer. Vervolgens word In kort uiteensetting van sosio-retoriese analise gegee. Dit is In multidimensionele en multidissiplinere benadering wat deur Vernon Robbins ontwikkel is en wat In teks as In hegte eenheid van verskillende teksture sien. In hierdie studie word hierdie metode gebruik, ofskoon met In paar aanpassings. Die makrostruktuur van Romeine word in hoofstuk 4 met behulp van In epistolere en retoriese analise nagegaan. Daarna word Rom. 7 as In retoriese eenheid binne Rom. 5-8 as die breer ko-teks van Rom.7 aangedui. Vervolgens word die retoriese situasie van Romeine asook bepaalde retoriese tegnieke en strategiee van Paulus bespreek. Die standpunt word ingeneem dat Romeine as deliberatiewe retoriek beskou moet word. In hoofstukke 5 tot 7 word die verskillende teksture van Rom. 7 aan die orde gestel. Die herhalend-progressiewe tekstuur, begin-middel-slot tekstuur en die argumentatiewe tekstuur word ondersoek om die interne tekstuur vas te stel. In Analise van die ethumeme in die teks lewer ook In bydrae om die vloei van die argument te kan volg. Hieruit volg dat Rom. 7:1-6 as In analogie beskou moet word, wat In belangrike rniddel in argumentasie was. Verder kan afgelei word dat die teenswoordige tydsvorm in Rom. 7: 14-25 In onderdeel is van die kombinasie van outobiografies-tipies-retoriese kenmerke wat in argumentasie gebruik word. In die bestudering van die intertekstuur word aandag aan die volgende voorbeelde van skriftelike intertekstuur gegee: resitasie (met weglatings) en die uitbou van In tema. By die bestudering van die kulturele intertekstuele analise kom voorbeelde van Joodse intertekstuur aan die orde: Rom. 7:8-10 as toespeling op Gen. 3 en Eks. 3, Paulus se gebruik van die "ek", die wet, die beeld van slawe en die bose begeerte. Verder kan Rom.7:15, 19 en die "ek" as toespelings op Griekse tragedies, die sonde as mag, en slawemy as Grieks-Romeinse kulturele intertekstuur gesien kan word. Wat die sosiale intertekstuur betref word bevind dat die analogie van die huwelik meer verwantskap met die Joodse huwelik as met die Grieks- Romeinse huwelik vertoon. Ten slotte word die teologiese tekstuur ondersoek. Eers word aandag gegee aan die heilsgeskiedenis en die verbond van God om In begrip van die teologiese wereld van Paulus te verkry. Daama word die hamartologie, antropologie en ten slotte die wet by Paulus ondersoek. Daar word bevind dat VOl-lOS" in Rom. 7 meestal die universele morele wet van God, geskrewe en ongeskrewe, en nie net die wet van Moses nie, aandui. In bepaalde gevalle, soos in Rom.7:21-25, beteken dit "beginsel" of reel." Rom.7 is in sy geheel'n verwerping van die beswaar of misverstand wat na aanleiding van Paulus se uitsprake oor die wet in die vorige hoofstukke kon ontstaan het. Paulus stel in Rom.7 die verhouding tussen gelowiges en die wet aan die orde asook die funksie van die wet met betrekking tot sonde by die onbekeerde. Op hierdie wyse ontken hy ten sterkste dat die wet sonde is terwyl hy ook die funksie van die wet met behulp van sy eie ervaring uitbeeld.

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