Therapeutische Erfolge von IVIG sind gut dokumentiert, aber die zu Grunde liegenden molekularen Mechanismen sind noch nicht vollständig erforscht. Molekulare Analysen unseres Labors über die Interaktion von IVIG mit Fabs von Patienten, die an einer autoimmunen Thrombozytopenie (ITP) leiden zeigten, dass die am häufigsten selektierten Fab von den V3-23 und V3-30 VH-Keimbahngenen abstammten. Eine weitere Studie mit IgG und IgM Phagen-Display Bibliotheken von einem gesunden Spender zeigten ebenfalls eine bevorzugte Reaktivierung von IVIG mit Fabs vom Ursprung der V3-23 und V3-30 Gene. Es konnte gefolgert werden, dass diese Interaktion von IVIG mit Fabs von diesen zwei VH-Genen weder alleine auf den Gesundheitsstatus des Spenders zurückzuführen war, noch auf eine zuvor erfolgte Behandlung mit IVIG. Diese Dissertation wurde unter Verwendung der Phagen-Display Technologie unternommen, um die molekulare Interaktion von IVIG mit Antikörpern zu erforschen, die von einem Patienten kloniert wurden, der an einem systemischen Lupus erythematodes und rheumatischem Fieber leidet. Die Resultate waren mit den früheren Studien zu vergleichen, insbesondere mit den Daten eines Patienten, der zu der ITP einen Lupus entwickelte. 23 Fabs, welche 7 unabhängige Klone repräsentierten, wurden isoliert. Im Gegensatz zu von Patienten mit ITP abstammenden Klonen reagierte keines von den in dieser Studie selektierten Fabs mit Thrombozyten. Die über IVIG gebundene Fab-Phagen stammten hierbei ausschließlich von den V3-23 und V3-30 VH-Genen ab. Darüber hinaus wurde beobachtet, dass von diesen Fabs verschiedene CDR3 Regionen einschließlich verschiedenen D- und JH-Gensegmenten benutzt wurden. Die Ergebnisse zeigten weiterhing, dass die Bindung von IVIG an die Fabs unabhängig von der Leichten Kette war. Ihrem Keimbahngen-Ursprung entsprechend hatten die Fabs Aminosäuren an Positionen in den FR1, FR3 und im 3'-Ende von CDR2, die dafür bekannt sind, dass sie für die Bindung des B-Zell-Superantigens Staphylococcus Protein A (SpA) essentiell sind. Es wurde gezeigt, dass sich zwar einige von den Fabs stark an SpA banden, aber keine Korrelation in der Intensität zur Bindung mit IVIG vorlag. Einige Fabs zeigten eine schwache Bindung an HIV gp120, einem anderen B-Zell-Superantigen. Zusammenfassend lässt sich aus der vorliegenden Studie und den vorherigen Ergebnissen schließen, dass ein Anteil von IVIG wie ein B-Zellen Superantigen funktionieren könnte, das für die Bildung und Regulation des normalen B-Zellen Repertoires wichtig ist. Der Bindungsmechanismus scheint ähnlich, aber nicht identisch mit dem der anderen getesteten B-Zellen-Superantigene zu sein. / The beneficial therapeutic effects of IVIG are well documented, but the underlying molecular mechanisms are not fully understood. Recent investigations from our laboratory into the molecular analysis of Fabs bound by IVIG from patients suffering from autoimmune thrombocytopenia revealed that the most frequently selected Fabs originated from the V3-23 and V3-30 VH germline genes. A subsequent study with IgG and IgM phage display libraries from a healthy donor also demonstrated a preferential reactivity of IVIG to Fabs of V3-23 and V3-30 origin. That study revealed that the unique reactivity of IVIG to Fabs of these two VH gene loci was not restricted to the autoimmune nature of the donors, neither to previous treatment with IVIG. One of the thrombocytopenia patients developed lupus. This study was undertaken to study the molecular interaction of IVIG with antibodies selected from a patient suffering from systemic lupus erythematosus and rheumatic fever using phage display technology, and to compare the results with the previous studies. Twenty-three Fabs representing seven independent clones were isolated. In contrast to ITP-derived clones, none of the Fabs selected in this study reacted with platelets. The Fab phages bound by IVIG were sequenced in order to determine their VH gene usage and clonal relatedness. V3-23 and V3-30 VH genes were found to be exclusively utilized by the Fab phages bound by IVIG. Moreover, different CDR3 regions including different D and JH gene segments were observed to be used by these Fabs. The results further showed that the binding of IVIG to the Fabs was independent of the light chain since different light chains were observed to be associated with the VH3 immunoglobulins. Detailed sequence analysis of the Fabs revealed the presence of amino acid residues at positions within FR1, FR3, and the 3' end of CDR2 that are known to be contacted by the B cell superantigen Staphylococcus protein A (SpA). Some of the Fabs were shown to bind strongly to SpA, but there was no correlation with the binding-intensity to IVIG. Some bound very weakly to HIV gp120, another B cell superantigen. This study, together with previous results, suggests that a subset of IVIG may function as a B cell superantigen that may significantly shape the B cell repertoire. The binding mechanism appears to be similar but not identical to the other tested B cell superantigens.

Thesis (MScMedSc (Pathology. Medical Microbiology))--University of Stellenbosch, 2010. / ENGLISH ABSTRACT: S. aureus causes serious infections in the hospital and community settings. The rate of MRSA infections are rapidly increasing worldwide. Currently, at Tygerberg hospital, approximately a third of S. aureus isolates are MRSA. This was the first epidemiological study of S. aureus conducted at Tygerberg Hospital that included prospective clinical data on patients with S. aureus bacteraemia together with spa typing of strains and the detection of the mecA and pvl genes in a multiplex PCR. Clonal cluster groups of S. aureus isolates were obtained by BURP analysis and compared to international important clones. The molecular epidemiology of hospital acquired (HA), health-care associated (HCA) and community acquired (CA) S. aureus bacteraemic strains at this hospital was examined. Lastly, repeat isolates of patients were collected to analyse any possible organism-related factors associated with persistent and recurrent bacteraemia. We investigated a total number of 113 S. aureus strains from 104 patients (70% MSSA, 30% MRSA). Repeat strains consisted of nine isolates (from 5 patients). All isolates were obtained from blood cultures collected during the period March 2008 to May 2009. Phenotypic and genotypic detection of methicillin resistance correlated well. According to the literature, most CA-MRSA strains are distinguishable from HA-MRSA strains based upon the presence of the PVL toxin. However, no CA-MRSA was detected in our study, therefore the association between HA-MRSA versus CA-MRSA strains could not be analysed. In this study, CA-MSSA was identified in 22% of all MSSA isolates versus 0% CA-MRSA. PVL positive strains were found in 22.7% of all MSSA isolates with no detection in MRSA isolates. It was noted that MRSA strains clustered in spa CC-701 and CC-012, whereas CC-002 only contained MSSA strains. Likewise HA-strains representing the majority of MRSA strains also clustered in spa CC-701 and CC-012. Forty nine spa types were identified in 89.3% of all isolates, whereas 9.7% of these strains were non-typeable. Five novel spa types were revealed. We detected a diverse number of spa-types that correlated to international clones. The most predominant spa type found in our setting was t037 (only in MRSA), followed by t891. According to the literature, t037 is associated to the Brazilian/Hungarian clone (SCCmec type III; ST 239). Our findings, as well as other South African studies, indicate that t037 has been identified in clinical strains from numerous provinces in South Africa. Interestingly, all isolates from spa type t891 were PVL positive MSSA. Bacteraemia cases were predominantly related to catheter sepsis, followed by skin and soft tissue infections (SSTI). Only one persistent bacteraemia case was identified related to a HA-SSTI. Recurrent bacteraemia cases were found in patients on dialysis for chronic renal failure and in burns patients related to intravascular catheter infections. The local epidemiology of S. aureus and the prevalence rate of different strains are important to investigate. The information provided contributes to the epidemiology of staphylococcal strains causing bacteraemia in our setting. These insights are useful for optimal diagnostic and therapeutic measures. The techniques developed can be used to identify outbreaks and recurrent infections. / AFRIKAANSE OPSOMMING: S. aureus veroorsaak ernstige infeksies in die hospitaalomgewing en in die gemeenskap. Wêreldwyd, neem metisillien-weerstandige S. aureus (MRSA) infeksies vinnig toe. Huidiglik by Tygerberg hospitaal is ongeveer ‘n derde van S. aureus isolate MRSA. Hierdie is die eerste epidemiologiese studie by Tygerberg hospitaal wat prospektiewe kliniese data van pasiënte met S. aureus bakteremie saam met spa tipering en aantoning van die mecA en pvl gene in ‘n multipleks PKR insluit. Klonale groepe (spa-CC) van MRSA en MSSA isolate is deur BURP analise verkry, en vergelyk met internasionaal belangrike klone. Die molekulêre epidemiologie van hospitaalverworwe (HA), gesondheidsorgverworwe (HCA) en gemeenskapsverworwe (CA) S. aureus bakteremie by hierdie hospitaal is ondersoek. Laastens, oorspronklike en daaropvolgende herhaal isolate is gekollekteer om moontlike organisme- faktore geassosieerd met persisterende en herhalende bakteremiese episodes te analiseer. Ons het in totaal 113 S. aureus isolate van 104 pasiënte ondersoek (70% MSSA, 30% MRSA). Nege isolate (van 5 pasiënte) was herhaal isolate. Alle isolate was afkomstig vanaf bloedkulture wat gedurende die periode Maart 2008 tot Mei 2009 gekollekteer is. Fenotipiese en genotipiese aantoning van metisillien weerstandigheid het goed gekorreleer. Volgens die literatuur kan die meeste CA-MRSA isolate van HA-MRSA isolate onderskei word op grond van die teenwoordigheid van die PVL toksien. Geen CA-MRSA is egter in ons studie gevind nie, dus kon die assosiasie tussen HA-MRSA en CA-MRSA isolate nie ondersoek word nie. CA-MSSA was in 22% van alle MSSA geidentifiseer teenoor 0% CA-MRSA. PVL is in MSSA isolate gevind (22.7% van alle MSSA) maar glad nie in MRSA nie. Dit is opgemerk dat MRSA isolate hoofsaaklik in spa CC 701 en CC-012 kloongroepe voorkom, teenoor kloongroep CC-002 wat slegs MSSA isolate bevat het. Soortgelyk het HA-isolate wat die meerderheid van MRSA isolate verteenwoordig het ook in kloongroepe 1 & 2 gegroepeer. Nege-en-veertig spa tipes is geïdentifiseer in 89.3% of alle isolate en 9.7% was nie-tipeerbaar. Vyf nuwe spa tipes is getoon. Ons het ‘n diverse aantal spa-tipes geïdentifiseer wat met internasionale klone gekorreleer het. Die mees dominante spa tipe in ons omgewing was t037 (slegs in MRSA), gevolg deur t891. Volgens die literatuur word t037 met die Brasiliaanse/Hongaarse kloon geassosieer (SCCmec tipe III; ST 239). Ons bevindings, asook ander Suid Afrikaanse studies, dui aan dat t037 in kliniese isolate vanaf talle provinsies in Suid-Afrika aangetoon is. Van belang is dat al die isolate van spa tipe t891 MSSA en PVL positief was. Bakteremiese gevalle was hoofsaaklik geassosieer met kateter-sepsis, gevolg deur vel en sagteweefsel infeksies (SSTI). Slegs een persisterende bakteremiese geval was geïdentifiseer geassosieer met HA-SSTI. Herhalende bakteremiese episodes is in pasiënte op dialise vir kroniese nierversaking en in brandwonde pasiënte met intra-vaskulêre kateter infeksies aangetoon. Die lokale epidemiologie van S. aureus en die prevalensie koers van verskillende stamme is van belang. Hierdie inligting dra by tot kennis van die epidemiologie van stafilokokkale stamme wat in ons omgewing bakteremie veroorsaak. Hierdie insigte is nuttig vir optimale diagnostiese en terapeutiese riglyne. Die tegnieke wat ontwikkel is, kan gebruik word om uitbrake en herhalende infeksies te identifiseer.

Darbo objektas: kurortų ir kurortinių teritorijų patrauklumo kriterijai. Darbo tikslas: išanalizuoti kokiais kriterijais remiantis klientai renkasi kurortą ar kurortinę teritoriją. Problema: nėra pakankamai informacijos apie kurortų ir kurortinių teritorijų vertinimo kriterijus. Darbo uždaviniai: 1. Atskleisti kurorto ir kurortinės teritorijos sampratą. 2. Išsiaiškinti paslaugų patrauklumo kriterijus klientams kurortuose ir kurortinėse teritorijose. 3. Atlikti kurortuose ir kurortinėse teritorijose nustatytų patrauklumo elementų vertinimą. Darbo rezultatai ir išvados: Straipsnyje analizuojama kurortų samprata ir aptariami kurortų statuso reikalavimai. Nustatyta, kad pagal teikiamų paslaugų specifiką dauguma Lietuvos kurortų (Druskininkai, Birštonas ir Palanga) yra priskiriami mišriam kurortų tipui ir tik vienas - Neringa - poilsio tipui. Straipsnyje aptartos 4 Lietuvos kurortinės teritorijos (Anykščiai, Trakai, Zarasai, Ignalinos miesto Strigailiškio ir Palūšės kaimų dalių teritorijos). Išskirti pagrindiniai kurortų ir kurortinių teritorijų patrauklumo kriterijai – infrastruktūra, kainos – kokybės santykis, pramogos, sveikatingumas. Nustatyta, kad labiausiai kurortų patrauklumas susijęs su infrastruktūra, taip pat įrodyta, kad ir kiti minėti vertinimo kriterijai yra svarbūs renkatis kurortą/kurortinę teritoriją. / Object of the study: resorts and spa areas attractiveness criteria. Purpose of the study: analyze what criteria customers choose a resort or spa areas. Problem: there is not enough information about resorts and spa areas in the evaluation criteria. Main goals of this study: 1. Reveal to the resorts and resort areas concept. 2. To find out the criteria for attractiveness to customers resorts and spa areas. 3. Perform resorts and spa areas attractive elements of the evaluation set. Results and conclusions. In this article is analyzed and discussed about the concept of resort and it‘s status requirements. Also, presented resorts classification. It was found that by it‘s specifics resorts in Lithuania are classified as mixed type resort (Palanga, Druskininkai, Birštonas) and rest resort – Neringa. In this article is discussed about 4 Lithuania resort areas (Anykščiai, Trakai, Zarasai, Strigailiškis and Palūšės countryside parts of Ignalina city). In this article is also distinguished the main resorts and resort areas attractiveness criteria - infrastructure, price - quality ratio, entertainment, wellness. It was found that the main criteria of resort attractiveness is it‘s infrastructure, but in this article is also mentioned that price – quality ratio, entertainment and wellness is also important for choosing resort.

The enzyme 11β-Hydroxysteroid Dehydrogenase 1 (11β-HSD1) catalyses the intracellular biosynthesis of the active glucocorticoid cortisol. Tissue specific dysregulation of the enzyme has been implicated in the development of metabolic syndrome and other associated diseases. Experiments with transgenic mice and prototype inhibitors show that inhibition of 11β-HSD1 in visceral adipose tissue and liver leads to a resistance of diet-induced hyperglycemia and a favourable lipid and lipoprotein profile as compared to controls. 11β-HSD1 inhibition has thus been proposed as an effective strategy to decrease intracellular glucocorticoid levels without affecting circulating glucocorticoid levels that are essential for stress responses. The clinical development of selective and potent drugs has therefore become a priority. In this research, a process of virtual screening employing the novel algorithm UFSRAT (Ultra Fast Shape Recognition with Atom Types) was used to discover compounds which had specific physicochemical and spatial atomic parameters deemed essential for inhibition of 11β-HSD1. The top scoring compounds were assayed for inhibitory activity against recombinant human and mouse enzyme, using a fluorescence spectroscopy approach. In addition, HEK-293 cell based assays with either human, mouse or rat enzymes were carried out using a scintillation proximity assay (SPA). The most potent compound competitively inhibited human 11β-HSD1 with a Kiapp value of 51 nM. Recombinant mouse and human enzyme were expressed, purified and characterised and used in a series of ligand binding assays. Further to this, an X-ray crystal structure of mouse 11β-HSD1 in complex with a tight binding inhibitor – carbenoxolone was solved.

The purpose of this research is to investigate the dispersion of ammonia (NH3) from three Concentrated Animal Feeding Operations (CAFOs) in western Kentucky, as well as to investigate the Weather Research and Forecasting – Chemistry (WRF-Chem) model’s sensitivity response to initial NH3 concentrations under both wet conditions (significant precipitation) and dry conditions (no precipitation). As expected, pollutant concentrations generally were significantly higher near their points of origin and generally declined away from the sources. Contrary to expectations, ammonia tended to rise through the planetary boundary layer (PBL) regardless of atmospheric conditions. Results showed modeled NH3 pollution levels at the surface generally to be higher under wet conditions. A GIS-based analysis method was developed to investigate model sensitivity to initial NH3 concentrations. Using this method, it was found that WRF-Chem exhibits an exponential relation between initial NH3 concentration and the final amount of NH3 produced by the model.

El presente estudio; descriptivo, no experimental y transversal, tiene como objetivo aportar conocimientos en relación a la evaluación del dolor miofascial, a través de la determinación del umbral de dolor a la presión de los músculos Masetero y Temporal en este tipo de pacientes, en sujetos sanos y la posible relación entre dichos umbrales y el género de los sujetos.

El estudio es descriptivo de tipo transversal, no experimental, abarcando una muestra de 1600 fichas clínicas de personas mayores de 15 años obtenidas mediante muestreo aleatorio simple en el CESFAM “Cristo Vive”. Los resultados indicaron que las EMERs más prevalentes, en el centro de salud en referencia, fueron el Lumbago no especificado con una prevalencia global de 10,25%; lesión de hombro no especificada (Hombro doloroso) 4,05% y Gonartrosis (osteoartrosis de rodilla) 2,56%. Las hipótesis planteadas (H1; H2; H3) resultaron rechazadas al no encontrarse una asociación estadística significativa (p>0.05) entre las variables involucradas, aceptando las hipótesis H0 correspondientes a cada una, de no relación entre las variables: EMER más prevalentes y nivel educacional. Los datos de prevalencia hallados en la muestra estudiada, fueron cercanos a la bibliografía revisada, en otros casos, la prevalencia fue menor que la esperada, probablemente a que se consideró solo el año 2004 como criterio de inclusión.

Thèse numérisée par la Direction des bibliothèques de l'Université de Montréal.

A b s t r a c t Title: Payment of Royalties for Performing the Broadcast in the Bedrooms of Spa Facilities The purpose of this thesis is to analyse the issue of payment of royalties for performing broadcast in the bedrooms of spa facilities, namely by the form of case study. The introduction shortly presents chosen subject and highlights the inconguity of jurisprudence in legal disputes relating to payment of royalties by spa facilities. Chapter one is dedicated to historical progression of different versions of the exceptions contained in section 23 of the Czech Copyrigt Act, which is also the cause for spa facilities to conclude that they are not liable for paying royalties to appropriate collecting society. Chapter two examines relavant jurisprudence by Regional Courts, High Courts and the Supreme Court of the Czech Republic. The jurisprudence of the Court of Justice of the European Union is also mentioned. Chapter three is composed of the introduction to the case study which illustrates the circumstances of the origin of hypothetic legal dispute which is to be solved by this thesis. Chapter four is the focal point of the thesis and it is presented by the judgement issued by a Regional Court in the legal dispute between the collecting society as a plaintiff and the spa facility as a respondent. The...

This thesis describes the importance of spa tourism in Greece reflecting on its history as well as its current status in Greek tourism and the health industry. Its content features basic terminology of the spa industry, specifications and development of the European balneology. This thesis also offers a view on Greek spa market and its performance including legislative and legal framework of the spa sector as well as a SWOT analysis of the Greek spa market. Last but not least it focuses on prospects and possibilities of cooperation between Greece and The Czech Republic in this field. It does so by introducing a new product to the travel industry for Czech clients. This new product takes into consideration current state of demand and health service in the Czech Republic as well as the offerings available from the Greek side.