Staphylococcus aureus (S. aureus) är en av många bakterier som har utvecklat resistens mot olika typer av antibiotika. På mitten av 1940-talet detekterades penicillinresistent S. aureus. Meticillinresistent S. aureus (MRSA) identifierades sedan år 1961. Vankomycin, en tricyklisk hydrofil glykopeptid, används fortfarande som ett förstahandsläkemedel mot allvarliga MRSA-infektioner trots förekomsten av vankomycinresistens och tillgången till nya MRSA-läkemedel. Syftet med detta arbete är att jämföra effektiviteten av vankomycin med andra läkemedel vid behandlingen av MRSA. Detta görs för att kunna bedöma om användningen av vankomycin som ett förstahandsläkemedel mot MRSA bör omprövas. Detta arbete är en litteraturstudie som är gjord med hjälp av artiklar från databasen PubMed. Det hittades fem studier om behandlingen av olika MRSA-infektioner. Linezolid visade sig vara effektivare än vankomycin vid behandlingen av hud- och mjukdelsinfektioner samt nosokomial pneumoni. Daptomycin var inte underlägset vankomycin mot bakteriemi och endokardit. Arbekacin visade sig vara mindre effektiv än vankomycin mot kronisk mediaotit. Kombinationen trimetoprim/ sulfametoxazol (TMP/SMX) verkade vara överlägset vankomycin vid behandlingen av hälsovårdsförvärvad pneumoni och ventilatorförvärvad pneumoni. Den slutsats som kan dras i detta arbete är att vid en jämförelse av vankomycin med linezolid, daptomycin, TMP/SMX och arbekacin vid behandling av olika MRSA-infektioner var vankomycin överlag mindre effektiv. Det kan därför vara lämpligt att ompröva ifall vankomycin bör användas som ett förstahandsläkemedel mot MRSA.

Charles University, Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Study program: Bioanalytical laboratory diagnostics in health care Candidate: Bc. Martina Malíková Thesis supervisor: PharmDr. Ondřej Janďourek, Ph.D. Consultant: RNDr. Irena Hanovcová, CSc. Title of diploma thesis: Effectiveness of disinfectants for hygienic hand disinfection against clinically important strains of enterococci Alcohol-based hand disinfectants for hygienic hand disinfection are a key tool for the control of nosocomial infections worldwide. The aim of this study was to verify the effectiveness of these products, which are used in healthcare facilities, against vancomycin-sensitive and vancomycin-resistant enterococci isolated from clinical materials. Testing of the efficacy of disinfectants was performed according to the Czech technical standard ČSN EN 1040 - Chemical disinfectants and antiseptics - Quantitative test using suspension to determine the basic bactericidal effect of chemical disinfectants and antiseptics - Test method and requirements (phase 1). We used the dilution method with a neutraliser for our testing. There were used a total of 35 strains of enterococci to determine efficacy, 12 of which were vancomycin sensitive (four E. faecium strains and eight E. faecalis...

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V průběhu projektu byly vyvinuty unikátní kolagenní pěny z kolagenu získaného z kůže sladkovodní ryby (kapr obecný, Cyprinus carpio). Pomocí síťování karbodiimidem byl překonán problém s nestabilitou kolagenní matrix z kolagenu získávaného z chladnokrevných živočichů při tělesné teplotě savců. Následně byly pěny impregnovány antibiotiky (gentamicin a vankomycin) a opětovně lyofilizovány, což je postup, který zajišťuje požadovanou koncentraci antibiotika bez rizika následného vymytí při dalších technologických krocích. Uvedený produkt je, na rozdíl od přípravků z nesíťovanéhokolagenu, stabilní i při sterilizaci gamma zářením. Finální sterilizovaný produkt byl testován in vivo na potkaním modelu infikované rány. Byla prokázána efektivita v léčbě potenciálně letální infekce Pseudomonas aeruginosa a kmene Stafylococcus aureus rezistentní k meticilinu (MRSA). Vzhledem k vysoké potřebě profylaxe a terapie infekcí pooperačních a jiných ran právě výše uvedenými polyrezistentními původci se jedná o slibný prostředek k budoucímu klinickému využití. Zkušenosti, které jsme získali v průběhu uvolnování ATB z kolagenních pěn budou v dalším vyvoji použity pro impregnaci zevní kolagenní vrstvy cévní protézy, čímž bychom mohli eliminovat jednu z největších nevýhod a rizik spojených s použitím umělých materiálu a tím je...

Glycopeptides are the so-called last-resort antibiotics in clinical practice used to treat heavier, predominantly nosocomial infections caused by multi-resistant coagulase-negative staphylococci. The origin and genetic basis of resistance to glycopeptide antibiotics has not yet been elucidated within coagulase-negative staphylococci. Research on Staphylococcus aureus has shown, that intermediate resistance to glycopeptide antibiotics is associated with the presence of one or more mutations, rather than being conditioned by the support of a particular genetic element, such as in enterococci. By using various types of in vitro resistant mutant selection, we were able to obtain isogenic pairs of glycopeptide sensitive and resistant strains of Staphylococcus epidermidis and Staphylococcus haemolyticus. By sequencing the genomes of these pairs, one nucleotide polymorphisms were identified and predominantly found in metabolic and cell wall control systems. Phenotypic analysis did not reveal a direct association of glycopeptide resistance with increased biofilm formation. In clinical practice, the cross-resistance of glycopeptides and other antibiotics is problematic. For the non-glycopeptide antibiotics imipenem and rifampicin, the incidence of cross-resistance with glycopeptide antibiotics in S. aureus...

Panther Fusion® system är ett fullständigt automatiserat in vitro diagnostiksystem som har högt flöde av prov-till-resultat och tillåter utförandet av in vitro diagnostiska tester genom användning av realtids Polymerase Chain Reaction (realtids-PCR). Syftet med detta projekt är att validera om det finns möjlighet att använda ett kit från Amplidiag på Panther fusion open access för att diagnostisera vankomycin resistenta enterokocker (VRE) samt att utvärdera analys direkt på E-swab prover istället för anrikningsbuljong. Det är viktigt att kunna detektera vanA-och vanB-generna i prover för att VRE hyser dessa gener. Sensitivitet och effektivitet av Panther fusion open access undersöktes genom att analysera proverna parallellt med Bio-Rad CFX384/96-PCR som är rutinmetoden för VRE-diagnostik på Klinisk mikrobiologi i Lund. Specificitet av en tidigare publicerad PCR-mix (PCR-mix) som kan detektera vanA och vanB i prover jämfördes med Amplidiag assay mix 2 som används i rutinmetoden. För att kunna utvärdera skillnaden av specificiteten och känsligheten mellan E-swab och anrikningsbuljong utfördes analysen på både E-swab och anrikningsbulong med både Panther fusion open access och Bio-Rad CFX384/96-PCR. MgCl2- koncentration optimerades till 3,0 mM. Sensitivitet och effektivitet av Panther fusion open access var högre med PCR-mix än Amplidiag assay mix 2 och uppvisade optimalt linjäritet. Specificitetstest av PCR-mixen visade att den bara kunde detektera vanA och vanB. Resultaten av spikade VRE-negativa patientprover (E-swab) visade sig vara orimliga då flera av proverna och kontrollerna gav resultat för båda generna. För att ersätta VRE-diagnostik på Bio-Rad CFX384/96 med Panther fusion open access behöver ytterligare experiment genomföras direkt på E-swab och vankomycine variabla enterokocker (VVE)-prover på Panther fusion open access. / Panther Fusion® system is a fully automated in vitro diagnostic system that has a high flow of sample-to-results and allows the performance of in vitro diagnostics tests using real-time Polymerase Chain Reaction (real-time PCR). The purpose of this project is to validate whether it is possible to use a kit from Amplidiag on Panther fusion open access to diagnose vancomycin resistant enterococcus (VRE). In addition, this project aims to evaluate and analyze E-swab samples directly instead of enrichment broth. It is important to be able to detect VanA and VanB genes in samples because VRE harbors these genes. Sensitivity and efficiency of Panther fusion open access were tested by analyzing the samples simultaneously with Bio-Rad CFX384/96 which is the routine method for VRE diagnostic at Clinical Microbiology in Lund. Specificity of a previously published PCR-mix (PCR-mix) that can detect VanA and VanB genes in samples was compared with Amplidiag assay mix 2 used in the routine method. In order to evaluate the difference in specificity and sensitivity between E-swab and enrichment broth, the analysis was performed on both E-swab and enrichment broth with both Panther fusion open access and Bio-Rad CFX384/96-PCR. MgCl2 concentration was optimized to 3,0 mM. Both sensitivity and efficiency of Panther fusion open access were higher with PCR-mix than those with Amplidiag assay mix 2 and it showed optimal linearity. Specificity test of PCR-mix detected only VanA and VanB genes. The results of inoculated the negative VRE-patient samples (E-swab) indicated that they were unreasonable because some of the samples and controls gave results of both genes. Replacement of the VRE-diagnostic method on Bio-Rad CFX384/96 with Panther fusion open access requires performing additional experiments directly on E-swab and vancomycin variable enterococcus (VVE) samples using Panther fusion open access.

A membrane protein VanZTei which is encoded by the gene vanZ from the vanA glycopeptide resistance gene cluster is a part of the large family of VanZ proteins. VanZTei confers resistance to teicoplanin in Enterococcus faecalis without the presence of other proteins encoded by the cluster. The aim of my work was to compare the ability of two orthologous proteins VanZTei and VanZg (from the genome of Enterococcus faecium) to confer resistance to glycopeptides in Staphylococcus aureus RN4220 and Enterococcus faecium. We have shown that VanZg increases resistance to teicoplanin (Tei) 8 to 16 times the and also to dalbavancin (Dalb) 8 times. VanZTei also confers resistance to Tei and Dalb, but the increase is only twofold. Conversely VanZTei confers resistance to newly synthetized glycopeptides more effectively than VanZg (fourfold increase of resistance confered by VanZTei and two to fourfold increase of resistance confered by VanZg). It suggests that both proteins have different specificity to antibiotics. In despite the mutants of S. aureus RN4220 VanZTei pRMC2 with increased resistance to teicoplanin (MICTei> 8 µg/ml) in which the resistance is dependent on vanZTei expression were selected. These resistant mutants do not carry mutation in a gene vanZTei or in its ribosomal binding site. Neither of the...

In this diploma work is check possibility application indirect photometric detection for enantiomers separation lactic acids and beta-hydroxybutyric acid with capillary electrophoresis. As chiral selector is used vankomycin. Verifyed his ability partial separation lactic acid on antipode with using picric acid as vizualition anion in background electrolyte. Used was coated capillary. Application indirect photometric detection succeed. Chiral separation beta-hydroxybutyric acid wasn't successful. Hip roll by selection fit vizualization anions for purposes indirect photometric detection. Are measured their electrophoretic and spectrophotometric characteristics, for inquest compatibility use in capillary electrophoresis in relation to indirect photometric detection. From timed datums were to be you - nap optimal vizualizatin anions for others experimentation. According to ascertained and measured feature reflect be optimal eosine soluble in alcohol and bromophenol blue.

The thesis is focused on the research of stability of controlled drug release systems based on a blend of plasticized starch/polycaprolactone (TPS/PCL) that served as a carrier. Antibiotic vancomycin was used as a model drug, and its release from TPS/PCL pellets into aqueous environment was followed by UV-spectroscopy and the obtained time dependences were treated by a simple kinetic model. Moreover, the simultaneous release of starch particles to the surrounding liquid phase was studied by static and dynamic light scattering as well as transmission electron microscopy (TEM) in order to obtain information on the stability of biodegradable matrix and on the structure of the products of the pellet decomposition on a nanoscale level. Key words: vancomycin, starch, drug delivery system, polycaprolactone (PCL), particle release, dynamic light scattering (DLS), static light scattering (SLS)