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104. Activity of daptomycin- and vancomycin-loaded poly-epsilon-caprolactone microparticles against mature staphylococcal biofilms

Author

Ferreira,Inês, Bettencourt,Ana, Gonçalves,Lídia, Kasper,Stefanie, Bétrisey,Bertrand, Kikhney,Judith, Moter,Annette, Trampuz,Andrej, Almeida,António J, Ferreira,Inês, Bettencourt,Ana, Gonçalves,Lídia, Kasper,Stefanie, Bétrisey,Bertrand, Kikhney,Judith, Moter,Annette, Trampuz,Andrej, and Almeida,António J

Abstract

Inês Santos Ferreira,1 Ana F Bettencourt,1 Lídia MD Gonçalves,1 Stefanie Kasper,2 Bertrand Bétrisey,3 Judith Kikhney,2 Annette Moter,2 Andrej Trampuz,4 António J Almeida1 1Research Institute for Medicines (iMed.ULisboa), Faculty of Pharmacy, University of Lisbon, Lisbon, Portugal; 2Biofilmcenter, German Heart Institute Berlin, Berlin, Germany; 3Infectious Diseases Service, Department of Medicine, Centre Hospitalier Universitaire Vaudois, Lausanne, Switzerland; 4Center for Musculoskeletal Surgery, Charité – University Medicine Berlin, Berlin, Germany Abstract: The aim of the present study was to develop novel daptomycin-loaded poly-epsilon-caprolactone (PCL) microparticles with enhanced antibiofilm activity against mature biofilms of clinically relevant bacteria, methicillin-resistant Staphylococcus aureus (MRSA) and polysaccharide intercellular adhesin-positive Staphylococcus epidermidis. Daptomycin was encapsulated into PCL microparticles by a double emulsion-solvent evaporation method. For comparison purposes, formulations containing vancomycin were also prepared. Particle morphology, size distribution, encapsulation efficiency, surface charge, thermal behavior, and in vitro release were assessed. All formulations exhibited a spherical morphology, micro­meter size, and negative surface charge. From a very early time stage, the released concentrations of daptomycin and vancomycin were higher than the minimal inhibitory concentration and continued so up to 72 hours. Daptomycin presented a sustained release profile with increasing concentrations of the drug being released up to 72 hours, whereas the release of vancomycin stabilized at 24 hours. The antibacterial activity of the microparticles was assessed by isothermal microcalorimetry against planktonic and sessile MRSA and S. epidermidis. Regarding planktonic bacteria, daptomycin-loaded PCL microparticles presented the highest antiba

Published
2015

105. In situ Verfahren zum molekularbiologischen Erregernachweis und zur Analyse medizinisch relevanter Biofilme

Author

Moter, Annette

Subjects
tropheryma whipplei, molecular diagnostics, sepsis, PCR, FISH, microscopy, endocarditis, 600 Technik, Medizin, angewandte Wissenschaften::610 Medizin und Gesundheit, periodontitis, biofilm
Abstract

Die wichtigste Aufgabe der medizinischen Mikrobiologie ist die Isolierung und Identifizie¬rung humanpathogener Organismen. Für die korrekte Wahl der Antibiotikatherapie und somit für die Prognose des Patienten ist eine schnelle, sensitive und spezifische Erregerdiagnostik von größter Wichtigkeit. Dabei sind die kulturellen Verfahren mit Anzucht der Infektionserreger auf festen oder in flüssigen Nährmedien zur Isolierung und Anreicherung individueller Spezies mit nachfolgender biochemischer Differenzierung ein fester Bestandteil der Routinediagnostik. Bereits kurz nach Probeneingang kann ein gefärbtes Direktpräparat wertvolle Hinweise für die initiale Therapieempfehlung geben. Nach erfolgreicher Anzucht folgt nach ein bis drei Tagen eine präzisere Erregerdifferenzierung. Viele Mikroorganismen sind jedoch schwer oder bisher nicht kultivierbar. Die Nährstoffbedürfnisse dieser anspruchsvollen Spezies sind zu spezifisch und komplex, um sie unter Laborbedingungen zu kultivieren. Andere Spezies wachsen zu langsam, um eine zeitnahe Erregeridentifikation zu ermöglichen. Auch für die Untersuchung von Probenmaterial von antibiotisch vorbehandelten Patienten sind kulturell basierte Techniken oft ungeeignet. Hier werden zunehmend molekularbiologische Verfahren eingesetzt, da sie das genetische Material der Mikroorganismen kulturunabhängig nachweisen. Ein Nachteil der meisten molekularbiologischen Methoden ist es jedoch, dass sie keine Information über räumliche Verteilung der Mikroorganismen zueinander oder im Kontext mit dem Gewebe erlauben. Somit sind insbesondere die hochsensitiven Nukleinsäure-Amplifikationstechniken kontaminationsanfällig und positive Nachweise von Krankheitserregern können nur unter Berücksichtigung klinischer Befunde interpretiert werden. Hier findet die Fluoreszenz in situ Hybridisierung (FISH) ihre Anwendung, bei welcher fluoreszenz-markierte Oligonukleotidsonden spezifisch an komplementäre RNA-Sequenzen im Bakterium binden. Sie erlaubt somit die Visualisierung und gleichzeitig kulturunabhängige Identifikation von Mikroorganismen in ihrem natürlichen Habitat und bildet somit die Brücke zwischen Molekularbiologie, Mikrobiologie und Pathologie. Die FISH ist ein viel versprechendes Verfahren, das sich zu einem wertvollen Instrument für die Analyse mikrobieller Biofilme sowie die mikrobiologische Routinediagnostik entwickelt hat. Sie schlägt eine Brücke zwischen Molekulargenetik und konventioneller Fluoreszenzmikroskopie und eignet sich damit auch für den Einsatz in den Laboratorien, die nicht die räumlichen, personellen und auch finanziellen Voraussetzungen für eine zuverlässige und kosteneffiziente molekulargenetische Diagnostik erfüllen. Der Einsatz der FISH für diagnostische Fragestellungen wurde am Beispiel der Blutkulturdiagnostik, der Endokarditis und der Mykobakteriosen diskutiert. Dabei erwies sich die Technik insbesondere bei Infektionen durch schwer kultivierbare Erreger wie Bartonella quintana, Tropheryma whipplei oder Mycobacterium leprae sowie nach antibiotischer Vorbehandlung als hilfreich. Durch die räumliche Auflösung kann die FISH Fragen beantworten, die derzeit mit keiner anderen Technik bearbeitet werden können. So liefert sie wertvolle Informationen für die Infektionsbiologie, wie die Erforschung komplexer Biofilme, wie am Beispiel der Parodontitis gezeigt. Weiterhin kann sie für Untersuchungen zur Relevanz schwer kultivierbarer und nicht kultivierter Mikroorganismen, aber auch für funktionelle Analysen, wie Bakterium-Wirt Interaktionen eingesetzt werden. Diese Eigenschaft wurde bei den Untersuchungen zur Dermatitis digitalis genutzt. Darüber hinaus gibt die FISH Auskunft zum rRNA Gehalt einer einzelnen bakteriellen Zelle und somit ihrer Aktivität in situ, wie bei den Biofilmen der infektiösen Endokarditis demonstriert. Es ist daher denkbar, dass sogar Vitalitätsmessungen von Bakterien in Biofilmen möglich sind, was neue Perspektiven der in situ Analyse der Biofilm Regulation eröffnet., The ultimate goal of medical microbiology is the fast and specific detection of pathogens. Conventional microbiological diagnosis is mainly based on culture-dependent methods that often fail due to previous antibiotic therapy or the involvement of fastidious or slowly growing microorganisms. During the last years molecular techniques entered the field of routine diagnostics. These mainly amplification based techniques are culture independent and highly sensitive, but they carry the risk of false positive results and do not give information about spatial distribution of the bacteria, their interaction with the host, or bacterial community structures. It is generally accepted that the favoured lifestyle of bacteria in natural environments is the biofilm. Medical biofilms are present on a wide range of interfaces in the human body. Whereas biofilms have been studied extensively in vitro, we are just beginning to learn about their structure and function in vivo because models fail to display the high complexity of the microenvironments in the human body. Direct detection of microorganisms in diagnostic specimens by fluorescence in situ hybridization (FISH) allows identification of the causative agent and simultaneous visualization within the histological context. We used FISH for fast identification of Gram-positive cocci in blood cultures, for detection of slowly growing bacteria, like mycobacteria in tissue sections, and for diagnosis of fastidious organisms, like Bartonella quintana or Tropheryma whipplei in heart valve tissues of endocarditis patients. Also yet uncultured species like Mycobacterium leprae or oral treponemes could be visualized in tissue sections. Furthermore, FISH was used to visualize and quantify amount, composition, and interactions of in vivo grown biofilms. The architecture of multispecies biofilms was analyzed using digital image analysis of oral biofilms showing the in vivo co-localization of two pairs of microorganisms on a single cell level. This approach can also be employed to investigate the abundance and localisation of not yet cultivable microorganisms. The tissue invasiveness of treponemes at the biofilm-host interface was shown for digital dermatitis. On heart valves of endocarditis patients we detected structured biofilms that were highly organized and similar to biofilms described in in vitro systems. The endocarditis biofilms showed stratification with zones of elevated rRNA levels alternating with layers of only DAPI positive cells that were presumably dead. We also found FISH positive bacteria in culture negative samples and samples from patients under antibiotic therapy, indicating recent metabolic activity of the cells. In summary, these findings show the versatility and impact of FISH for biofilm infections and diagnostics in the clinical setting.

Published
2011
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110. Contribution of Veillonella parvula to Pseudomonas aeruginosa-Mediated Pathogenicity in a Murine Tumor Model System

Author

Pustelny, Christian, primary, Komor, Uliana, additional, Pawar, Vinay, additional, Lorenz, Anne, additional, Bielecka, Agata, additional, Moter, Annette, additional, Gocht, Benjamin, additional, Eckweiler, Denitsa, additional, Müsken, Mathias, additional, Grothe, Claudia, additional, Lünsdorf, Heinrich, additional, Weiss, Siegfried, additional, and Häussler, Susanne, additional

Published
2015
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111. Fluorescence in situ hybridization (FISH) in the microbiological diagnostic routine laboratory: a review.

Author

Frickmann, Hagen, Zautner, Andreas Erich, Moter, Annette, Kikhney, Judith, Hagen, Ralf Matthias, Stender, Henrik, and Poppert, Sven

Subjects
INFECTION, FLUORESCENCE in situ hybridization, ANTIBIOTICS, DRUG resistance, MICROSCOPY, DIAGNOSIS
Abstract

Early identification of microbial pathogens is essential for rational and conservative antibiotic use especially in the case of known regional resistance patterns. Here, we describe fluorescencein situhybridization (FISH) as one of the rapid methods for easy identification of microbial pathogens, and its advantages and disadvantages for the diagnosis of pathogens in human infections in the laboratory diagnostic routine. Binding of short fluorescence-labeled DNA or nucleic acid-mimicking PNA probes to ribosomes of infectious agents with consecutive analysis by fluorescence microscopy allows identification of bacterial and eukaryotic pathogens at genus or species level. FISH analysis leads to immediate differentiation of infectious agents without delay due to the need for microbial culture. As a microscopic technique, FISH has the unique potential to provide information about spatial resolution, morphology and identification of key pathogens in mixed species samples. On-going automation and commercialization of the FISH procedure has led to significant shortening of the time-to-result and increased test reliability. FISH is a useful tool for the rapid initial identification of microbial pathogens, even from primary materials. Among the rapidly developing alternative techniques, FISH serves as a bridging technology between microscopy, microbial culture, biochemical identification and molecular diagnostic procedures. [ABSTRACT FROM AUTHOR]

Published
2017
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112. Effects of Diode Laser, Gaseous Ozone, and Medical Dressings on Enterococcus faecalis Biofilms in the Root Canal Ex Vivo.

Author

Bitter, Kerstin, Vlassakidis, Alexander, Niepel, Mediha, Hoedke, Daniela, Schulze, Julia, Neumann, Konrad, Moter, Annette, and Noetzel, Jörn

Subjects
BACTERIAL disease prevention, INFECTION prevention, ANALYSIS of variance, ANTI-infective agents, BIOFILMS, DENTAL care, DENTAL materials, DENTISTRY, ENDODONTICS, GASES, IRRIGATION (Medicine), MEDICAL care, MEDICAL technology, OZONE, PATIENTS, ROOT canal treatment, SALT, DENTAL extraction, CALCIUM compounds, ENTEROCOCCAL infections, CONTROL groups, DATA analysis software, KRUSKAL-Wallis Test, ANTIMICROBIAL bandages, COLONY-forming units assay, DIAGNOSIS
Abstract

The objective was to compare the antibacterial effects of adjunctive disinfection using diode laser and gaseous ozone compared to the medical dressings calcium hydroxide (Ca(OH)2) and chlorhexidine gel (CHX-Gel) on Enterococcus faecalis biofilms in human root canals ex vivo. Root canals of 180 human extracted teeth were infected by E. faecalis and divided into 3 main groups (G): G1, control; G2, instrumentation and irrigation using 0.9% NaCl; G3, instrumentation and irrigation using 1% NaOCl. In each main group, the following treatments were applied: gaseous ozone, diode laser, and medical dressings of Ca(OH)2 or CHX-Gel for 7 days (n=15). Reduction of colony forming units (CFUs) inside the root canal of planktons and frequencies of adherent bacteria after treatment were calculated. Bacterial reduction was significantly affected by the irrigation protocol (p<0.0005) and the disinfection method (p<0.0005), and a significant interaction between both factors could be observed (p<0.0005; ANOVA). In G3 (instrumentation using 1% NaOCl), no significant effect of disinfection methods could be demonstrated on planktonic bacteria (p=0.062; ANOVA) and frequencies of adherent bacteria (p>0.05; chi-square test). Instrumentation and irrigation using NaOCl combined with ozone or laser application resulted in comparable bacterial reduction on E. faecalis to the application of medical dressings. [ABSTRACT FROM AUTHOR]

Published
2017
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114. Towards diagnostic guidelines for biofilm-associated infections

Author

Hall-Stoodley, Luanne, Stoodley, Paul, Kathju, Sandeep, Høiby, Niels, Moser, Claus, William Costerton, J, Moter, Annette, Bjarnsholt, Thomas, Hall-Stoodley, Luanne, Stoodley, Paul, Kathju, Sandeep, Høiby, Niels, Moser, Claus, William Costerton, J, Moter, Annette, and Bjarnsholt, Thomas

Abstract

Biofilms associated with the human body, particularly in typically sterile locations, are difficult to diagnose and treat effectively because of their recalcitrance to conventional antibiotic therapy and host immune responses. The study of biofilms in medicine today requires a translational approach, with examination of clinically relevant biofilms in the context of specific anatomic sites, host tissues, and diseases, focusing on what can be done to mitigate their pathologic consequences. This review, which grew out of a discussion session on clinical biofilms at the 5th ASM Biofilm Conference in Cancun, Mexico, is designed to give an overview of biofilm-associated infections (BAI) and to propose a platform for further discussion that includes clinicians, medical microbiologists, and biofilm researchers who are stakeholders in advancing the scientific pursuit of better diagnosis and treatment of BAI to mitigate their human and healthcare costs. It also highlights the need for better diagnostic markers, which exploit the difference between planktonic and biofilm cells.

Published
2012

117. Immunopathology of Immune Reconstitution Inflammatory Syndrome in Whipple’s Disease

Author

Moos, Verena, primary, Feurle, Gerhard E., additional, Schinnerling, Katina, additional, Geelhaar, Anika, additional, Friebel, Julian, additional, Allers, Kristina, additional, Moter, Annette, additional, Kikhney, Judith, additional, Loddenkemper, Christoph, additional, Kühl, Anja A., additional, Erben, Ulrike, additional, Fenollar, Florence, additional, Raoult, Didier, additional, and Schneider, Thomas, additional

Published
2013
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121. High Frequency of Tropheryma whipplei in Culture-Negative Endocarditis

Author

Geißdörfer, Walter, primary, Moos, Verena, additional, Moter, Annette, additional, Loddenkemper, Christoph, additional, Jansen, Andreas, additional, Tandler, René, additional, Morguet, Andreas J., additional, Fenollar, Florence, additional, Raoult, Didier, additional, Bogdan, Christian, additional, and Schneider, Thomas, additional

Published
2012
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122. A Novel Mouse Model of Staphylococcus aureusVascular Graft Infection

Author

Van de Vyver, Hélène, Bovenkamp, Philipp R., Hoerr, Verena, Schwegmann, Katrin, Tuchscherr, Lorena, Niemann, Silke, Kursawe, Laura, Grosse, Christina, Moter, Annette, Hansen, Uwe, Neugebauer, Ute, Kuhlmann, Michael T., Peters, Georg, Hermann, Sven, and Löffler, Bettina

Abstract

Staphylococcus aureuscauses very serious infections of vascular grafts. Knowledge of the molecular mechanisms of this disease is largely lacking because of the absence of representable models. Therefore, the aim of this study was to set up a mouse model of vascular graft infections that closely mimics the human situation. A catheter was inserted into the right carotid artery of mice, which acted as a vascular graft. Mice were infected i.v. using 8 different S. aureusstrains, and development of the infection was followed up. Although all strains had varying abilities to form biofilm in vitroand different levels of virulence in mice, they all caused biofilm formation on the grafts. This graft infection was monitored using magnetic resonance imaging (MRI) and 18F-fluordeoxyglucose positron emission tomography (FDG-PET). MRI allowed the quantification of blood flow through the arteries, which was decreased in the catheter after infection. FDG-PET revealed high inflammation levels at the site of the catheter after infection. This model closely resembles the situation in patients, which is characterized by a tight interplay between pathogen and host, and can therefore be used for the testing of novel treatment, diagnosis, and prevention strategies. In addition, combining MRI and PET with microscopic techniques provides an appropriate way to characterize the course of these infections and to precisely analyze biofilm development.

Published
2017
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124. Molecular Characterization of Borrelia persica, the Agent of Tick Borne Relapsing Fever in Israel and the Palestinian Authority

Author

Safdie, Gracia, primary, Farrah, Iba Y., additional, Yahia, Reem, additional, Marva, Esther, additional, Wilamowski, Amos, additional, Sawalha, Samer S., additional, Wald, Naama, additional, Schmiedel, Judith, additional, Moter, Annette, additional, Göbel, Ulf B., additional, Bercovier, Herve, additional, Abdeen, Ziad, additional, Assous, Marc V., additional, and Fishman, Yolanta, additional

Published
2010
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125. Filifactor alocis - involvement in periodontal biofilms

Author

Schlafer, Sebastian, primary, Riep, Birgit, additional, Griffen, Ann L, additional, Petrich, Annett, additional, Hübner, Julia, additional, Berning, Moritz, additional, Friedmann, Anton, additional, Göbel, Ulf B, additional, and Moter, Annette, additional

Published
2010
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134. Gram-Negative Bacteria Aggravate Murine Small Intestinal Th1-Type Immunopathology following Oral Infection with Toxoplasma gondii

Author

Heimesaat, Markus M., primary, Bereswill, Stefan, additional, Fischer, André, additional, Fuchs, David, additional, Struck, Daniela, additional, Niebergall, Julia, additional, Jahn, Hannah-Katharina, additional, Dunay, Ildikò R., additional, Moter, Annette, additional, Gescher, Dorothee M., additional, Schumann, Ralf R., additional, Göbel, Ulf B., additional, and Liesenfeld, Oliver, additional

Published
2006
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139. The Genus Treponema.

Author

Dworkin, Martin, Falkow, Stanley, Rosenberg, Eugene, Schleifer, Karl-Heinz, Stackebrandt, Erko, Norris, Steven J., Paster, Bruce J., Moter, Annette, and Göbel, Ulf B.

Published
2006
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146. In VivoEfficacy of Antimicrobials against Biofilm-Producing Pseudomonas aeruginosa

Author

Pawar, Vinay, Komor, Uliana, Kasnitz, Nadine, Bielecki, Piotr, Pils, Marina C., Gocht, Benjamin, Moter, Annette, Rohde, Manfred, Weiss, Siegfried, and Häussler, Susanne

Abstract

ABSTRACTPatients suffering from cystic fibrosis (CF) are commonly affected by chronic Pseudomonas aeruginosabiofilm infections. This is the main cause for the high disease severity. In this study, we demonstrate that P. aeruginosais able to efficiently colonize murine solid tumors after intravenous injection and to form biofilms in this tissue. Biofilm formation was evident by electron microscopy. Such structures could not be observed with transposon mutants, which were defective in biofilm formation. Comparative transcriptional profiling of P. aeruginosaindicated physiological similarity of the bacteria in the murine tumor model and the CF lung. The efficacy of currently available antibiotics for treatment of P. aeruginosa-infected CF lungs, such as ciprofloxacin, colistin, and tobramycin, could be tested in the tumor model. We found that clinically recommended doses of these antibiotics were unable to eliminate wild-type P. aeruginosaPA14 while being effective against biofilm-defective mutants. However, colistin-tobramycin combination therapy significantly reduced the number of P. aeruginosaPA14 cells in tumors at lower concentrations. Hence, we present a versatile experimental system that is providing a platform to test approved and newly developed antibiofilm compounds.

Published
2015
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147. Co-Localized or Randomly Distributed? Pair Cross Correlation of In Vivo Grown Subgingival Biofilm Bacteria Quantified by Digital Image Analysis.

Author

Schillinger, Claudia, Petrich, Annett, Lux, Renate, Riep, Birgit, Kikhney, Judith, Friedmann, Anton, Wolinsky, Lawrence E., Göbel, Ulf B., Daims, Holger, and Moter, Annette

Subjects
DENTAL plaque, PATHOGENIC microorganisms, DIGITAL images, IMAGE analysis, BACTERIA, BIOFILMS
Abstract

The polymicrobial nature of periodontal diseases is reflected by the diversity of phylotypes detected in subgingival plaque and the finding that consortia of suspected pathogens rather than single species are associated with disease development. A number of these microorganisms have been demonstrated in vitro to interact and enhance biofilm integration, survival or even pathogenic features. To examine the in vivo relevance of these proposed interactions, we extended the spatial arrangement analysis tool of the software daime (digital image analysis in microbial ecology). This modification enabled the quantitative analysis of microbial co-localization in images of subgingival biofilm species, where the biomass was confined to fractions of the whole-image area, a situation common for medical samples. Selected representatives of the diseaseassociated red and orange complexes that were previously suggested to interact with each other in vitro (Tannerella forsythia with Fusobacterium nucleatum and Porphyromonas gingivalis with Prevotella intermedia) were chosen for analysis and labeled with specific fluorescent probes via fluorescence in situ hybridization. Pair cross-correlation analysis of in vivo grown biofilms revealed tight clustering of F. nucleatum/periodonticum and T. forsythia at short distances (up to 6 μm) with a pronounced peak at 1.5 μm. While these results confirmed previous in vitro observations for F. nucleatum and T. forsythia, random spatial distribution was detected between P. gingivalis and P. intermedia in the in vivo samples. In conclusion, we successfully employed spatial arrangement analysis on the single cell level in clinically relevant medical samples and demonstrated the utility of this approach for the in vivo validation of in vitro observations by analyzing statistically relevant numbers of different patients. More importantly, the culture-independent nature of this approach enables similar quantitative analyses for "as-yet-uncultured" phylotypes which cannot be characterized in vitro. [ABSTRACT FROM AUTHOR]

Published
2012
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148. Impaired Immune Functions of Monocytes and Macrophages in Whipple's Disease.

Author

Moos, Verena, Schmidt, Carsten, Geelhaar, Anika, Kunkel, Désirée, Allers, Kristina, Schinnerling, Katina, Loddenkemper, Christoph, Fenollar, Florence, Moter, Annette, Raoult, Didier, Ignatius, Ralf, and Schneider, Thomas

Subjects
WHIPPLE'S disease, MACROPHAGE activation, MONOCYTES, FLOW cytometry, INTERLEUKINS, IMMUNOHISTOCHEMISTRY, TUMOR necrosis factors, PATIENTS
Abstract

Background & Aims: Whipple''s disease is a chronic multisystemic infection caused by Tropheryma whipplei. Host factors likely predispose for the establishment of an infection, and macrophages seem to be involved in the pathogenesis of Whipple''s disease. However, macrophage activation in Whipple''s disease has not been studied systematically so far. Methods: Samples from 145 Whipple''s disease patients and 166 control subjects were investigated. We characterized duodenal macrophages and lymphocytes immunohistochemically and peripheral monocytes by flow cytometry and quantified mucosal and systemic cytokines and chemokines indicative for macrophage activation. In addition, we determined duodenal nitrite production and oxidative burst induced by T whipplei and by other bacteria. Results: Reduced numbers of duodenal lymphocytes, increased numbers of CD163+ and stabilin-1+, reduced numbers of inducible nitric synthase+ duodenal macrophages, and increased percentages of CD163+ peripheral monocytes indicated a lack of inflammation and a M2/alternatively activated macrophage phenotype in Whipple''s disease. Incubation with T whipplei in vitro enhanced the expression of CD163 on monocytes from Whipple''s disease patients but not from control subjects. Chemokines and cytokines associated with M2/alternative macrophage activation were elevated in the duodenum and the peripheral blood from Whipple''s disease patients. Functionally, Whipple''s disease patients showed a reduced duodenal nitrite production and reduced oxidative burst upon incubation with T whipplei compared with healthy subjects. Conclusions: The lack of excessive local inflammation and alternative activation of macrophages, triggered in part by the agent T whipplei itself, may explain the hallmark of Whipple''s disease: invasion of the intestinal mucosa with macrophages incompetent to degrade T whipplei. [Copyright &y& Elsevier]

Published
2010
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149. Let them fly or light them up: matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry and fluorescencein situhybridization (FISH).

Author

Schweickert, Birgitta, Moter, Annette, Lefmann, Michael, and Göbel, Ulf B.

Subjects
*DIAGNOSTIC microbiology, *MEDICAL microbiology, *COMMUNICABLE disease diagnosis, *MATRIX-assisted laser desorption-ionization, *MASS spectrometry
Abstract

Schweickert B, Moter A, Lefmann M, Göbel UB. Let them fly or light them up: matrix-assisted laser desorption/Ionization time of flight (MALDI-TOF) mass spectrometry and fluorescencein situhybridization (FISH). APMIS 2004;112:856–85.This review focuses on clinical bacteriology and by and large does not cover the detection of fungi, viruses or parasites. It discusses two completely different but complementary approaches that may either supplement or replace classic culture-based bacteriology. The latter view may appear provocative in the light of the actual market penetration of molecular genetic testing in clinical bacteriology. Despite its elegance, high specificity and sensitivity, molecular genetic diagnostics has not yet reached the majority of clinical laboratories. The reasons for this are manifold: Many microbiologists and medical technologists are more familiar with classical microbiological methods than with molecular biology techniques. Culture-based methods still represent the work horse of everyday routine. The number of available FDA-approved molecular genetic tests is limited and external quality control is still under development. Finally, it appears difficult to incorporate genetic testing in the routine laboratory setting due to the limited number of samples received or the lack of appropriate resources. However, financial and time constraints, particularly in hospitals as a consequence of budget cuts and reduced length of stay, lead to a demand for significantly shorter turnaround times that cannot be met by culture-dependent diagnosis. As a consequence, smaller laboratories that do not have the technical and personal equipment required for molecular genetic amplification techniques may adopt alternative methods such as fluorescencein situhybridization (FISH) that combines easy-to-perform molecular hybridization with microscopy, a technique familiar to every microbiologist. FISH is hence one of the technologies presented here. For large hospital or reference laboratories with a high sample volume requiring massive parallel high-throughput testing we discuss matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF) of nucleic acids, a technology that has evolved from the post-genome sequencing era, for high-throughput sequence variation analysis (1, 2). [ABSTRACT FROM AUTHOR]

Published
2004
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150. Contribution of Veillonella parvulato Pseudomonas aeruginosa-Mediated Pathogenicity in a Murine Tumor Model System

Author

Pustelny, Christian, Komor, Uliana, Pawar, Vinay, Lorenz, Anne, Bielecka, Agata, Moter, Annette, Gocht, Benjamin, Eckweiler, Denitsa, Müsken, Mathias, Grothe, Claudia, Lünsdorf, Heinrich, Weiss, Siegfried, and Häussler, Susanne

Abstract

ABSTRACTThe recent finding that high numbers of strict anaerobes are present in the respiratory tract of cystic fibrosis (CF) patients has drawn attention to the pathogenic contribution of the CF microbiome to airway disease. In this study, we investigated the specific interactions of the most dominant bacterial CF pathogen, Pseudomonas aeruginosa, with the anaerobic bacterium Veillonella parvula, which has been recovered at comparable cell numbers from the respiratory tract of CF patients. In addition to growth competition experiments, transcriptional profiling, and analyses of biofilm formation by in vitrostudies, we used our recently established in vivomurine tumor model to investigate mutual influences of the two pathogens during a biofilm-associated infection process. We found that P. aeruginosaand V. parvulacolonized distinct niches within the tumor. Interestingly, significantly higher cell numbers of P. aeruginosacould be recovered from the tumor tissue when mice were coinfected with both bacterial species than when mice were monoinfected with P. aeruginosa. Concordantly, the results of in vivotranscriptional profiling implied that the presence of V. parvulasupports P. aeruginosagrowth at the site of infection in the host, and the higher P. aeruginosaload correlated with clinical deterioration of the host. Although many challenges must be overcome to dissect the specific interactions of coinfecting bacteria during an infection process, our findings exemplarily demonstrate that the complex interrelations between coinfecting microorganisms and the immune responses determine clinical outcome to a much greater extent than previously anticipated.

Published
2014
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