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Flow cytometry antibiotic

Nadine Czekalski, Stefanie Imminger, Elisabeth Salhi, Marjan Veljkovic, Karolin Kleffel, David Drissner, Frederik Hammes, Helmut Bürgmann, Urs von Gunten
Ozone, a strong oxidant and disinfectant, seems ideal to cope with future challenges of water treatment, such as micropollutants, multiresistant bacteria (MRB) and even intracellular antibiotic resistance genes (ARG), but information on the latter is scarce. In ozonation experiments we simultaneously determined kinetics and dose-dependent inactivation of Escherichia coli and its plasmid-encoded sulfonamide resistance gene sul1 in different water matrixes. Effects in E. coli were compared to an autochthonous wastewater community...
October 24, 2016: Environmental Science & Technology
Leise Riber, Mette Burmølle, Martin Alm, Stefan M Milani, Peter Thomsen, Lars H Hansen, Søren J Sørensen
The spread of antimicrobial resistance, usually mediated by horizontal transfer of plasmids, limits the options of treating bacterial infections and thereby poses a crucial human health problem. The disturbance of plasmid stability within bacterial species in clinical environments serves as a novel strategy to reduce the development and dissemination of antibiotic resistance. We tested the ability of irgasan to destabilize plasmids from Escherichia coli K-12 cells when added directly into liquid growth medium at concentrations below levels of marked bacterial growth inhibition, or when released into liquid growth medium from irgasan-impregnated Interpenetrating Polymer Network (IPN) silicone hydrogel objects, a novel technology developed as drug-delivery platform...
October 12, 2016: Plasmid
Wanida Phetsang, Ruby Pelingon, Mark S Butler, Sanjaya Kc, Miranda E Pitt, Geraldine Kaeslin, Matthew A Cooper, Mark A T Blaskovich
Reduced susceptibility to antimicrobials in Gram-negative bacteria may result from multiple resistance mechanisms, including increased efflux pump activity or reduced porin protein expression. Up-regulation of the efflux pump system is closely associated with multidrug resistance (MDR). To help investigate the role of efflux pumps on compound accumulation, a fluorescence-based assay was developed using fluorescent derivatives of trimethoprim (TMP), a broad-spectrum synthetic antibiotic that inhibits an intracellular target, dihydrofolate reductase (DHFR)...
October 14, 2016: ACS Infectious Diseases
Walid Mohamed, Eugen Domann, Trinad Chakraborty, Gopala Mannala, Katrin S Lips, Christian Heiss, Reinhard Schnettler, Volker Alt
BACKGROUND: Staphylococcus aureus is the principle causative pathogen of osteomyelitis and implant-associated bone infections. It is able to invade and to proliferate inside osteoblasts thus avoiding antibiotic therapy and the host immune system. Therefore, development of alternative approaches to stimulate host innate immune responses could be beneficial in prophylaxis against S. aureus infection. TLR9 is the intracellular receptor which recognizes unmethylated bacterial CpG-DNA and activates immune cells...
October 3, 2016: BMC Microbiology
Song-Tao Han, Ying Fei, Jin-You Huang, Mei Xu, Li-Chan Chen, D Joshua Liao, Yu-Jie Tan
Extended-spectrum β-lactamase (ESBL) genes that render bacteria resistant to antibiotics are commonly detected using phenotype testing, which is time consuming and not sufficiently accurate. To establish a better method, we used phenotype testing to identify ESBL-positive bacterial strains and conducted PCR to screen for TEM (named after the patient Temoneira who provided the first sample), sulfhydryl reagent variable (SHV), cefotaxime (CTX)-M-1, and CTX-M-9, the 4 most common ESBL types and subtypes. We then performed multiplex PCR with 1 primer containing a biotin and hybridized the PCR products with gene-specific probes that were coupled with microbeads and coated with a specific fluorescence...
September 16, 2016: Journal of Biomolecular Techniques: JBT
Petra Hildebrandt, Kristin Surmann, Manuela Gesell Salazar, Nicole Normann, Uwe Völker, Frank Schmidt
Staphylococcus aureus is a Gram-positive opportunistic pathogen that is able to cause a broad range of infectious diseases in humans. Furthermore, S. aureus is able to survive inside nonprofessional phagocytic host cell which serve as a niche for the pathogen to hide from the immune system and antibiotics therapies. Modern OMICs technologies provide valuable tools to investigate host-pathogen interactions upon internalization. However, these experiments are often hampered by limited capabilities to retrieve bacteria from such an experimental setting...
October 2016: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
Moritz Fritzenwanker, Can Imirzalioglu, Trinad Chakraborty, Florian M Wagenlehner
Urinary tract infections (UTIs) are among the most common bacterial infections and constitute for a considerable number of all antibiotic prescriptions. The surge of multi-resistant bacteria has significantly complicated empirical treatment leading to wide use of broad-spectrum antibiotics, further driving resistance development. New ways to detect and quantify host and bacterial molecules in urine promise opportunities to improve diagnostics of UTIs, which can translate to new approaches in treatment regimens...
November 2016: Expert Review of Anti-infective Therapy
B Martínez-Vallespín, W Vahjen, J Zentek
Medium-chain fatty acids (MCFAs) have been suggested as an alternative to the use of antibiotics in animal nutrition with promising results. First, we studied the sensitivity of Salmonella Enteritidis and an enteropathogenic Escherichia coli strain against caprylic (C8), capric (C10) and lauric (C12) acids. A porcine in vitro model using the porcine cell line IPEC-J2 was used to test the effects of MCFAs on structural and immunological traits without and with a concomitant challenge with E. coli or S. Enteritidis...
October 2016: Cytotechnology
Meital Gury-BenAri, Christoph A Thaiss, Nicolas Serafini, Deborah R Winter, Amir Giladi, David Lara-Astiaso, Maayan Levy, Tomer Meir Salame, Assaf Weiner, Eyal David, Hagit Shapiro, Mally Dori-Bachash, Meirav Pevsner-Fischer, Erika Lorenzo-Vivas, Hadas Keren-Shaul, Franziska Paul, Alon Harmelin, Gérard Eberl, Shalev Itzkovitz, Amos Tanay, James P Di Santo, Eran Elinav, Ido Amit
Innate lymphoid cells (ILCs) are critical modulators of mucosal immunity, inflammation, and tissue homeostasis, but their full spectrum of cellular states and regulatory landscapes remains elusive. Here, we combine genome-wide RNA-seq, ChIP-seq, and ATAC-seq to compare the transcriptional and epigenetic identity of small intestinal ILCs, identifying thousands of distinct gene profiles and regulatory elements. Single-cell RNA-seq and flow and mass cytometry analyses reveal compartmentalization of cytokine expression and metabolic activity within the three classical ILC subtypes and highlight transcriptional states beyond the current canonical classification...
August 25, 2016: Cell
F Ghasemi, R Jalal
Acinetobacter baumannii is a serious concern amongst hospitalised patients worldwide and its resistance to antibiotics has emerged as a threat to public health in recent years. Metal oxide nanoparticles were found to be effective for overcoming bacterial resistance owing to their antibacterial activities. The aim of this study was to investigate the combined effects of zinc oxide nanoparticles (ZnO-NPs) and the conventional antibiotics ciprofloxacin and ceftazidime as well as their mechanisms of action against resistant A...
September 2016: Journal of Global Antimicrobial Resistance
Claude Saint-Ruf, Steve Crussard, Christine Franceschi, Sylvain Orenga, Jasmine Ouattara, Mahendrasingh Ramjeet, Jérémy Surre, Ivan Matic
Rapidly treating infections with adequate antibiotics is of major importance. This requires a fast and accurate determination of the antibiotic susceptibility of bacterial pathogens. The most frequently used methods are slow because they are based on the measurement of growth inhibition. Faster methods, such as PCR-based detection of determinants of antibiotic resistance, do not always provide relevant information on susceptibility, particularly that which is not genetically based. Consequently, new methods, such as the detection of changes in bacterial physiology caused by antibiotics using flow cytometry and fluorescent viability markers, are being explored...
2016: Frontiers in Microbiology
Jeyce K F Andrade, Márcia I F Souza, Manoel A Gomes Filho, Diogo M F Silva, André L S Barros, Maria D Rodrigues, Paulo B N Silva, Silene C Nascimento, Jaciana S Aguiar, Dalci J Brondani, Gardênia C G Militão, Teresinha G Silva
BACKGROUND: Nitrofurantoin is a nitroderivative antibiotic that has bactericidal activity against pathogens causing urinary tract infection. A few studies have reported that nitrofurantoin has cytotoxic activity against cancer cells; however, nitrofurans remain a poorly explored class of compounds with respect to their anticancer potential. The aim of this study was to investigate the anticancer effects of a nitrofurantoin derivative, n-pentyl-nitrofurantoin (NFP), on HL-60 leukemia cells...
October 2016: Pharmacological Reports: PR
Karin Sá Fernandes, Paulo Sérgio da Silva Santos, Nathalie Pepe Medeiros de Rezende, Marina Gallottini
Kostmann syndrome (KS) is an autosomal recessive disorder characterized by a low neutrophil count and recurrent bacterial infections, including periodontal disease. This report describes the case of a 5-year-old female KS patient treated with regular infusions of granulocyte stimulating factor (granulokine), with primary complaint of gingival bleeding and tooth mobility, and followed up for 10 years. Conventional periodontal treatment and some extractions were performed, after neutrophil function tests using flow cytometry...
August 3, 2016: Special Care in Dentistry
Parveen Parasar, Amanda Wilhelm, Heloisa M Rutigliano, Aaron J Thomas, Lihong Teng, Bi Shi, William C Davis, Carlos E Suarez, Daniel D New, Kenneth L White, Christopher J Davies
Major histocompatibility complex class I (MHC-I) proteins can be expressed as cell surface or secreted proteins. To investigate whether bovine non-classical MHC-I proteins are expressed as cell surface or secreted proteins, and to assess the reactivity pattern of monoclonal antibodies with non-classical MHC-I isoforms, we expressed the MHC proteins in murine P815 and human K562 (MHC-I deficient) cells. Following antibiotic selection, stably transfected cell lines were stained with H1A or W6/32 antibodies to detect expression of the MHC-I proteins by flow cytometry...
August 2016: Research in Veterinary Science
Jinxin Zhao, Soon-Ee Cheah, Kade D Roberts, Roger L Nation, Philip E Thompson, Tony Velkov, Zongjun Du, Matthew D Johnson, Jian Li
Polymyxin B and colistin are exclusively active against Gram-negative pathogens and have been used in the clinic as a last-line therapy. In this study, we investigated the antimicrobial activity of a novel polymyxin, FADDI-019, against Staphylococcus aureus. MIC and time-kill assays were employed to measure the activity of FADDI-019 against S. aureus ATCC 700699. Cell morphology was examined with scanning electron microscopy (SEM), and cell membrane polarity was measured using flow cytometry. Transcriptome changes caused by FADDI-019 treatment were investigated using transcriptome sequencing (RNA-Seq)...
July 2016: MSphere
Hong Jiang, Meimei Xiong, Qiuyan Bi, Ying Wang, Chong Li
Considering that some antibacterial agents can identify the outer structure of pathogens like cell wall and/or cell membrane, we explored a self-enhanced targeted delivery strategy by which a small amount of the antibiotic molecules were modified on the surface of carriers as targeting ligands of certain bacteria while more antibiotic molecules were loaded inside the carriers, and thus has the potential to improve the drug concentration at the infection site, enhance efficacy and reduce potential toxicity. In this study, a novel targeted delivery system against methicillin-resistant Staphylococcus aureus (MRSA) pneumonia was constructed with daptomycin, a lipopeptide antibiotic, which can bind to the cell wall of S...
July 2016: Acta Pharmaceutica Sinica. B
G-J M van de Geijn, S Denker, V Meuleman-van Waning, H G M Koeleman, E Birnie, G-J Braunstahl, T L Njo
INTRODUCTIONS: Discriminating bacterial from nonbacterial acute exacerbations of chronic obstructive pulmonary disease (AECOPD) is difficult, causing antibiotics overuse and bacterial resistance. Sputum cultures are of limited use because results take time. In our hospital, only leukocyte concentration and CRP are laboratory parameters evaluated in AECOPD. We evaluated additional tests to discriminate bacterial vs. nonbacterial AECOPD: 5-part leukocyte differentiation (hematology analyzer), leukocyte differentiation using flow cytometry (Leukoflow, Cytodiff), Leuko64 kit, and procalcitonin...
July 27, 2016: International Journal of Laboratory Hematology
Faqing Tian, Chunyan Wang, Meiqin Tang, Juheng Li, Xiaohui Cheng, Sihan Zhang, Delan Ji, Yingcai Huang, Huiqing Li
Chloramphenicol is an old antibiotic that also inhibits mammalian mitochondrial protein synthesis. Our studies demonstrated that chloramphenicol is highly cytotoxic to myeloma cells, acting in a dose- and time-dependent manner. Chloramphenicol sharply suppressed ATP levels in myeloma cells at concentrations ≥ 25 μg/mL. Colorimetric and clonogenic assays indicate that chloramphenicol inhibits growth of myeloma cell lines at concentrations ≥ 50 μg/mL, and inhibits primary myeloma cell growth at concentrations ≥ 25 μg/mL...
July 16, 2016: Oncotarget
Andrew Conway Morris, Deepankar Datta, Manu Shankar-Hari, Christopher J Weir, Jillian Rennie, Jean Antonelli, Adriano G Rossi, Noel Warner, Jim Keenan, Alice Wang, K Alun Brown, Sion Lewis, Tracey Mare, A John Simpson, Gillian Hulme, Ian Dimmick, Timothy S Walsh
INTRODUCTION: Critically ill patients are at high risk of nosocomial infections, with between 20% and 40% of patients admitted to the intensive care unit (ICU) acquiring infections. These infections result in increased antibiotic use, and are associated with morbidity and mortality. Although critical illness is classically associated with hyperinflammation, the high rates of nosocomial infection argue for an importance of effect of impaired immunity. Our group recently demonstrated that a combination of 3 measures of immune cell function (namely neutrophil CD88, monocyte HLA-DR and % regulatory T cells) identified a patient population with a 2...
2016: BMJ Open
Gao Tianqin, Cheng Chunlei, Wang Jingjing
Neocarzinostatin (NCS) is a member of enediyne antibiotics with high anticancer potential. Our study was performed to explore the synergistic anti-glioma effects of NCS and paclitaxel (PTX) in vitro and in vivo. By MTT assay, the cytotoxicities of the drugs to human glioma cells U87MG and rat glioma cells C6 were evaluated. The results showed that the combinations of NCS and PTX can synergistically inhibit glioma cells survival. Cell apoptosis was detected by flow cytometry, and the results showed that the combinations of NCS and PTX synergistically enhanced apoptosis ratio of glioma cells...
July 14, 2016: Biological & Pharmaceutical Bulletin
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