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Sample records for obligate intracellular bacterial

  1. Proteomic Profiling of the Outer Membrane Fraction of the Obligate Intracellular Bacterial Pathogen Ehrlichia ruminantium

    PubMed Central

    Moumène, Amal; Marcelino, Isabel; Ventosa, Miguel; Gros, Olivier; Lefrançois, Thierry; Vachiéry, Nathalie

    2015-01-01

    The outer membrane proteins (OMPs) of Gram-negative bacteria play a crucial role in virulence and pathogenesis. Identification of these proteins represents an important goal for bacterial proteomics, because it aids in vaccine development. Here, we have developed such an approach for Ehrlichia ruminantium, the obligate intracellular bacterium that causes heartwater. A preliminary whole proteome analysis of elementary bodies, the extracellular infectious form of the bacterium, had been performed previously, but information is limited about OMPs in this organism and about their role in the protective immune response. Identification of OMPs is also essential for understanding Ehrlichia’s OM architecture, and how the bacterium interacts with the host cell environment. First, we developed an OMP extraction method using the ionic detergent sarkosyl, which enriched the OM fraction. Second, proteins were separated via one-dimensional electrophoresis, and digested peptides were analyzed via nano-liquid chromatographic separation coupled with mass spectrometry (LC-MALDI-TOF/TOF). Of 46 unique proteins identified in the OM fraction, 18 (39%) were OMPs, including 8 proteins involved in cell structure and biogenesis, 4 in transport/virulence, 1 porin, and 5 proteins of unknown function. These experimental data were compared to the predicted subcellular localization of the entire E. ruminantium proteome, using three different algorithms. This work represents the most complete proteome characterization of the OM fraction in Ehrlichia spp. The study indicates that suitable subcellular fractionation experiments combined with straightforward computational analysis approaches are powerful for determining the predominant subcellular localization of the experimentally observed proteins. We identified proteins potentially involved in E. ruminantium pathogenesis, which are good novel targets for candidate vaccines. Thus, combining bioinformatics and proteomics, we discovered new OMPs for E. ruminantium that are valuable data for those investigating new vaccines against this organism. In summary, we provide both pioneering data and novel insights into the pathogenesis of this obligate intracellular bacterium. PMID:25710494

  2. Adaptive immunity to the obligate intracellular pathogen Coxiella burnetii.

    PubMed

    Shannon, Jeffrey G; Heinzen, Robert A

    2009-01-01

    Coxiella burnetii is an obligate intracellular bacterial pathogen that causes the zoonosis Q fever. While an effective whole-cell vaccine (WCV) against Q fever exists, the vaccine has limitations in being highly reactogenic in sensitized individuals. Thus, a safe and effective vaccine based on recombinant protein antigen (Ag) is desirable. To achieve this goal, a better understanding of the host response to primary infection and the precise mechanisms involved in protective immunity to C. burnetii are needed. This review summarizes our current understanding of adaptive immunity to C. burnetii with a focus on recent developments in the field. PMID:18813881

  3. Microsporidian genome analysis reveals evolutionary strategies for obligate intracellular growth

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Microsporidia comprise a large phylum of obligate intracellular eukaryotes that are fungalrelated parasites responsible for widespread disease, and here we address questions about microsporidia biology and evolution. We sequenced three microsporidian genomes from two species, Nematocida parisii and...

  4. Evolutionary Genomics of a Temperate Bacteriophage in an Obligate Intracellular Bacteria (Wolbachia)

    E-print Network

    Bordenstein, Seth

    Evolutionary Genomics of a Temperate Bacteriophage in an Obligate Intracellular Bacteria (Wolbachia, the temperate double-stranded DNA bacteriophage WO in Wolbachia persistently transfers between bacterial considered the paradigm of temperate bacteriophage evolution in free-living bacteria, it appears irrelevant

  5. Metabolic Interdependence of Obligate Intracellular Bacteria and Their Insect Hosts†

    PubMed Central

    Zientz, Evelyn; Dandekar, Thomas; Gross, Roy

    2004-01-01

    Mutualistic associations of obligate intracellular bacteria and insects have attracted much interest in the past few years due to the evolutionary consequences for their genome structure. However, much less attention has been paid to the metabolic ramifications for these endosymbiotic microorganisms, which have to compete with but also to adapt to another metabolism—that of the host cell. This review attempts to provide insights into the complex physiological interactions and the evolution of metabolic pathways of several mutualistic bacteria of aphids, ants, and tsetse flies and their insect hosts. PMID:15590782

  6. Improved Quantification, Propagation, Purification and Storage of the Obligate Intracellular Human Pathogen Orientia tsutsugamushi

    PubMed Central

    Giengkam, Suparat; Blakes, Alex; Utsahajit, Peemdej; Chaemchuen, Suwittra; Atwal, Sharanjeet; Blacksell, Stuart D.; Paris, Daniel H.; Day, Nicholas P. J.; Salje, Jeanne

    2015-01-01

    Background Scrub typhus is a leading cause of serious febrile illness in rural Southeast Asia. The causative agent, Orientia tsutsugamushi, is an obligate intracellular bacterium that is transmitted to humans by the bite of a Leptotrombidium mite. Research into the basic mechanisms of cell biology and pathogenicity of O. tsutsugamushi has lagged behind that of other important human pathogens. One reason for this is that O. tsutsugamushi is an obligate intracellular bacterium that can only be cultured in mammalian cells and that requires specific methodologies for propagation and analysis. Here, we have performed a body of work designed to improve methods for quantification, propagation, purification and long-term storage of this important but neglected human pathogen. These results will be useful to other researchers working on O. tsutsugamushi and also other obligate intracellular pathogens such as those in the Rickettsiales and Chlamydiales families. Methodology A clinical isolate of O. tsutsugamushi was grown in cultured mouse embryonic fibroblast (L929) cells. Bacterial growth was measured using an O. tsutsugamushi-specific qPCR assay. Conditions leading to improvements in viability and growth were monitored in terms of the effect on bacterial cell number after growth in cultured mammalian cells. Key results Development of a standardised growth assay to quantify bacterial replication and viability in vitro. Quantitative comparison of different DNA extraction methods. Quantification of the effect on growth of FBS concentration, daunorubicin supplementation, media composition, host cell confluence at infection and frequency of media replacement. Optimisation of bacterial purification including a comparison of host cell lysis methods, purification temperature, bacterial yield calculations and bacterial pelleting at different centrifugation speeds. Quantification of bacterial viability loss after long term storage and freezing under a range of conditions including different freezing buffers and different rates of freezing. Conclusions Here we present a standardised method for comparing the viability of O. tsutsugamushi after purification, treatment and propagation under various conditions. Taken together, we present a body of data to support improved techniques for propagation, purification and storage of this organism. This data will be useful both for improving clinical isolation rates as well as performing in vitro cell biology experiments. PMID:26317517

  7. Disrupting Protein Expression with Peptide Nucleic Acids Reduces Infection by Obligate Intracellular Rickettsia

    PubMed Central

    Pelc, Rebecca S.; McClure, Jennifer C.; Kaur, Simran J.; Sears, Khandra T.; Rahman, M. Sayeedur; Ceraul, Shane M.

    2015-01-01

    Peptide Nucleic Acids (PNAs) are single-stranded synthetic nucleic acids with a pseudopeptide backbone in lieu of the phosphodiester linked sugar and phosphate found in traditional oligos. PNA designed complementary to the bacterial Shine-Dalgarno or start codon regions of mRNA disrupts translation resulting in the transient reduction in protein expression. This study examines the use of PNA technology to interrupt protein expression in obligate intracellular Rickettsia sp. Their historically intractable genetic system limits characterization of protein function. We designed PNA targeting mRNA for rOmpB from Rickettsia typhi and rickA from Rickettsia montanensis, ubiquitous factors important for infection. Using an in vitro translation system and competitive binding assays, we determined that our PNAs bind target regions. Electroporation of R. typhi and R. montanensis with PNA specific to rOmpB and rickA, respectively, reduced the bacteria’s ability to infect host cells. These studies open the possibility of using PNA to suppress protein synthesis in obligate intracellular bacteria. PMID:25781160

  8. Rickettsia Phylogenomics: Unwinding the Intricacies of Obligate Intracellular Life

    PubMed Central

    Gillespie, Joseph J.; Williams, Kelly; Shukla, Maulik; Snyder, Eric E.; Nordberg, Eric K.; Ceraul, Shane M.; Dharmanolla, Chitti; Rainey, Daphne; Soneja, Jeetendra; Shallom, Joshua M.; Vishnubhat, Nataraj Dongre; Wattam, Rebecca; Purkayastha, Anjan; Czar, Michael; Crasta, Oswald; Setubal, Joao C.; Azad, Abdu F.; Sobral, Bruno S.

    2008-01-01

    Background Completed genome sequences are rapidly increasing for Rickettsia, obligate intracellular ?-proteobacteria responsible for various human diseases, including epidemic typhus and Rocky Mountain spotted fever. In light of phylogeny, the establishment of orthologous groups (OGs) of open reading frames (ORFs) will distinguish the core rickettsial genes and other group specific genes (class 1 OGs or C1OGs) from those distributed indiscriminately throughout the rickettsial tree (class 2 OG or C2OGs). Methodology/Principal Findings We present 1823 representative (no gene duplications) and 259 non-representative (at least one gene duplication) rickettsial OGs. While the highly reductive (?1.2 MB) Rickettsia genomes range in predicted ORFs from 872 to 1512, a core of 752 OGs was identified, depicting the essential Rickettsia genes. Unsurprisingly, this core lacks many metabolic genes, reflecting the dependence on host resources for growth and survival. Additionally, we bolster our recent reclassification of Rickettsia by identifying OGs that define the AG (ancestral group), TG (typhus group), TRG (transitional group), and SFG (spotted fever group) rickettsiae. OGs for insect-associated species, tick-associated species and species that harbor plasmids were also predicted. Through superimposition of all OGs over robust phylogeny estimation, we discern between C1OGs and C2OGs, the latter depicting genes either decaying from the conserved C1OGs or acquired laterally. Finally, scrutiny of non-representative OGs revealed high levels of split genes versus gene duplications, with both phenomena confounding gene orthology assignment. Interestingly, non-representative OGs, as well as OGs comprised of several gene families typically involved in microbial pathogenicity and/or the acquisition of virulence factors, fall predominantly within C2OG distributions. Conclusion/Significance Collectively, we determined the relative conservation and distribution of 14354 predicted ORFs from 10 rickettsial genomes across robust phylogeny estimation. The data, available at PATRIC (PathoSystems Resource Integration Center), provide novel information for unwinding the intricacies associated with Rickettsia pathogenesis, expanding the range of potential diagnostic, vaccine and therapeutic targets. PMID:19194535

  9. Targeting of a Chlamydial Protease Impedes Intracellular Bacterial Growth

    PubMed Central

    Paschen, Stefan A.; Vier, Juliane; Schauenburg, Linda; Rupp, Jan; Meyer, Thomas F.; Häcker, Georg; Heuer, Dagmar

    2011-01-01

    Chlamydiae are obligate intracellular bacteria that propagate in a cytosolic vacuole. Recent work has shown that growth of Chlamydia induces the fragmentation of the Golgi apparatus (GA) into ministacks, which facilitates the acquisition of host lipids into the growing inclusion. GA fragmentation results from infection-associated cleavage of the integral GA protein, golgin-84. Golgin-84-cleavage, GA fragmentation and growth of Chlamydia trachomatis can be blocked by the peptide inhibitor WEHD-fmk. Here we identify the bacterial protease chlamydial protease-like activity factor (CPAF) as the factor mediating cleavage of golgin-84 and as the target of WEHD-fmk-inhibition. WEHD-fmk blocked cleavage of golgin-84 as well as cleavage of known CPAF targets during infection with C. trachomatis and C. pneumoniae. The same effect was seen when active CPAF was expressed in non-infected cells and in a cell-free system. Ectopic expression of active CPAF in non-infected cells was sufficient for GA fragmentation. GA fragmentation required the small GTPases Rab6 and Rab11 downstream of CPAF-activity. These results define CPAF as the first protein that is essential for replication of Chlamydia. We suggest that this role makes CPAF a potential anti-infective therapeutic target. PMID:21990969

  10. Bacterial pathogens commandeer Rab GTPases to establish intracellular niches.

    PubMed

    Stein, Mary-Pat; Müller, Matthias P; Wandinger-Ness, Angela

    2012-12-01

    Intracellular bacterial pathogens deploy virulence factors termed effectors to inhibit degradation by host cells and to establish intracellular niches where growth and differentiation take place. Here, we describe mechanisms by which human bacterial pathogens (including Chlamydiae; Coxiella burnetii; Helicobacter pylori; Legionella pneumophila; Listeria monocytogenes; Mycobacteria; Pseudomonas aeruginosa, Salmonella enterica) modulate endocytic and exocytic Rab GTPases in order to thrive in host cells. Host cell Rab GTPases are critical for intracellular transport following pathogen phagocytosis or endocytosis. At the molecular level bacterial effectors hijack Rab protein function to: evade degradation, direct transport to particular intracellular locations and monopolize host vesicles carrying molecules that are needed for a stable niche and/or bacterial growth and differentiation. Bacterial effectors may serve as specific receptors for Rab GTPases or as enzymes that post-translationally modify Rab proteins or endosomal membrane lipids required for Rab function. Emerging data indicate that bacterial effector expression is temporally and spatially regulated and multiple virulence factors may act concertedly to usurp Rab GTPase function, alter signaling and ensure niche establishment and intracellular bacterial growth, making this field an exciting area for further study. PMID:22901006

  11. Exploring anti-bacterial compounds against intracellular Legionella.

    PubMed

    Harrison, Christopher F; Kicka, Sébastien; Trofimov, Valentin; Berschl, Kathrin; Ouertatani-Sakouhi, Hajer; Ackermann, Nikolaus; Hedberg, Christian; Cosson, Pierre; Soldati, Thierry; Hilbi, Hubert

    2013-01-01

    Legionella pneumophila is a ubiquitous fresh-water bacterium which reproduces within its erstwhile predators, environmental amoeba, by subverting the normal pathway of phagocytosis and degradation. The molecular mechanisms which confer resistance to amoeba are apparently conserved and also allow replication within macrophages. Thus, L. pneumophila can act as an 'accidental' human pathogen and cause a severe pneumonia known as Legionnaires' disease. The intracellular localisation of L. pneumophila protects it from some antibiotics, and this fact must be taken into account to develop new anti-bacterial compounds. In addition, the intracellular lifestyle of L. pneumophila may render the bacteria susceptible to compounds diminishing bacterial virulence and decreasing intracellular survival and replication of this pathogen. The development of a single infection cycle intracellular replication assay using GFP-producing L. pneumophila and Acanthamoebacastellanii amoeba is reported here. This fluorescence-based assay allows for continuous monitoring of intracellular replication rates, revealing the effect of bacterial gene deletions or drug treatment. To examine how perturbations of the host cell affect L. pneumophila replication, several known host-targeting compounds were tested, including modulators of cytoskeletal dynamics, vesicle scission and Ras GTPase localisation. Our results reveal a hitherto unrealized potential antibiotic property of the ?-lactone-based Ras depalmitoylation inhibitor palmostatin M, but not the closely related inhibitor palmostatin B. Further characterisation indicated that this compound caused specific growth inhibition of Legionella and Mycobacterium species, suggesting that it may act on a common bacterial target. PMID:24058631

  12. Autophagic clearance of bacterial pathogens: molecular recognition of intracellular microorganisms

    PubMed Central

    Mansilla Pareja, Maria Eugenia; Colombo, Maria I.

    2013-01-01

    Autophagy is involved in several physiological and pathological processes. One of the key roles of the autophagic pathway is to participate in the first line of defense against the invasion of pathogens, as part of the innate immune response. Targeting of intracellular bacteria by the autophagic machinery, either in the cytoplasm or within vacuolar compartments, helps to control bacterial proliferation in the host cell, controlling also the spreading of the infection. In this review we will describe the means used by diverse bacterial pathogens to survive intracellularly and how they are recognized by the autophagic molecular machinery, as well as the mechanisms used to avoid autophagic clearance. PMID:24137567

  13. Transient Transfection and Expression in the Obligate Intracellular Parasite Toxoplasma gondii

    NASA Astrophysics Data System (ADS)

    Soldati, Dominique; Boothroyd, John C.

    1993-04-01

    Toxoplasma gondii is a protozoan pathogen that produces severe disease in humans and animals. This obligate intracellular parasite provides an excellent model for the study of how such pathogens are able to invade, survive, and replicate intracellularly. DNA encoding chloramphenicol acetyltransferase was introduced into T. gondii and transiently expressed with the use of three vectors based on different Toxoplasma genes. The ability to introduce genes and have them efficiently and faithfully expressed is an essential tool for understanding the structure-function relation of genes and their products.

  14. Scaling of immune responses against intracellular bacterial infection

    PubMed Central

    Abdullah, Zeinab; Knolle, Percy A

    2014-01-01

    Macrophages detect bacterial infection through pattern recognition receptors (PRRs) localized at the cell surface, in intracellular vesicles or in the cytosol. Discrimination of viable and virulent bacteria from non-virulent bacteria (dead or viable) is necessary to appropriately scale the anti-bacterial immune response. Such scaling of anti-bacterial immunity is necessary to control the infection, but also to avoid immunopathology or bacterial persistence. PRR-mediated detection of bacterial constituents in the cytosol rather than at the cell surface along with cytosolic recognition of secreted bacterial nucleic acids indicates viability and virulence of infecting bacteria. The effector responses triggered by activation of cytosolic PRRs, in particular the RIG-I-induced simultaneous rapid type I IFN induction and inflammasome activation, are crucial for timely control of bacterial infection by innate and adaptive immunity. The knowledge on the PRRs and the effector responses relevant for control of infection with intracellular bacteria will help to develop strategies to overcome chronic infection. PMID:25225613

  15. Chromerid genomes reveal the evolutionary path from photosynthetic algae to obligate intracellular parasites.

    PubMed

    Woo, Yong H; Ansari, Hifzur; Otto, Thomas D; Klinger, Christen M; Kolisko, Martin; Michálek, Jan; Saxena, Alka; Shanmugam, Dhanasekaran; Tayyrov, Annageldi; Veluchamy, Alaguraj; Ali, Shahjahan; Bernal, Axel; del Campo, Javier; Cihlá?, Jaromír; Flegontov, Pavel; Gornik, Sebastian G; Hajdušková, Eva; Horák, Aleš; Janouškovec, Jan; Katris, Nicholas J; Mast, Fred D; Miranda-Saavedra, Diego; Mourier, Tobias; Naeem, Raeece; Nair, Mridul; Panigrahi, Aswini K; Rawlings, Neil D; Padron-Regalado, Eriko; Ramaprasad, Abhinay; Samad, Nadira; Tom?ala, Aleš; Wilkes, Jon; Neafsey, Daniel E; Doerig, Christian; Bowler, Chris; Keeling, Patrick J; Roos, David S; Dacks, Joel B; Templeton, Thomas J; Waller, Ross F; Lukeš, Julius; Oborník, Miroslav; Pain, Arnab

    2015-01-01

    The eukaryotic phylum Apicomplexa encompasses thousands of obligate intracellular parasites of humans and animals with immense socio-economic and health impacts. We sequenced nuclear genomes of Chromera velia and Vitrella brassicaformis, free-living non-parasitic photosynthetic algae closely related to apicomplexans. Proteins from key metabolic pathways and from the endomembrane trafficking systems associated with a free-living lifestyle have been progressively and non-randomly lost during adaptation to parasitism. The free-living ancestor contained a broad repertoire of genes many of which were repurposed for parasitic processes, such as extracellular proteins, components of a motility apparatus, and DNA- and RNA-binding protein families. Based on transcriptome analyses across 36 environmental conditions, Chromera orthologs of apicomplexan invasion-related motility genes were co-regulated with genes encoding the flagellar apparatus, supporting the functional contribution of flagella to the evolution of invasion machinery. This study provides insights into how obligate parasites with diverse life strategies arose from a once free-living phototrophic marine alga. PMID:26175406

  16. The Obligate Intracellular Parasite Toxoplasma gondii Secretes a Soluble Phosphatidylserine Decarboxylase*

    PubMed Central

    Gupta, Nishith; Hartmann, Anne; Lucius, Richard; Voelker, Dennis R.

    2012-01-01

    Toxoplasma gondii is an obligate intracellular parasite capable of causing fatal infections in immunocompromised individuals and neonates. Examination of the phosphatidylserine (PtdSer) metabolism of T. gondii reveals that the parasite secretes a soluble form of PtdSer decarboxylase (TgPSD1), which preferentially decarboxylates liposomal PtdSer with an apparent Km of 67 ?m. The specific enzyme activity increases by 3-fold during the replication of T. gondii, and soluble phosphatidylserine decarboxylase (PSD) accounts for ?20% of the total PSD, prior to the parasite egress from the host cells. Extracellular T. gondii secreted ?20% of its total PSD activity at 37 °C, and the intracellular Ca2+ chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N?,N?-tetraacetic acid tetrakis (acetoxymethyl ester) inhibited the process by 50%. Cycloheximide, brefeldin A, ionic composition of the medium, and exogenous PtdSer did not modulate the enzyme secretion, which suggests a constitutive discharge of a presynthesized pool of PSD in axenic T. gondii. TgPSD1 consists of 968 amino acids with a 26-amino acid hydrophobic peptide at the N terminus and no predicted membrane domains. Parasites overexpressing TgPSD1-HA secreted 10-fold more activity compared with the parental strain. Exposure of apoptotic Jurkat cells to transgenic parasites demonstrated interfacial catalysis by secreted TgPSD1 that reduced host cell surface exposure of PtdSer. Immunolocalization experiments revealed that TgPSD1 resides in the dense granules of T. gondii and is also found in the parasitophorous vacuole of replicating parasites. Together, these findings demonstrate novel features of the parasite enzyme because a secreted, soluble, and interfacially active form of PSD has not been previously described for any organism. PMID:22563079

  17. The genome of obligately intracellular Ehrlichia canis revealsthemes of complex membrane structure and immune evasion strategies

    SciTech Connect

    Mavromatis, K.; Kuyler Doyle, C.; Lykidis, A.; Ivanova, N.; Francino, P.; Chain, P.; Shin, M.; Malfatti, S.; Larimer, F.; Copeland,A.; Detter, J.C.; Land, M.; Richardson, P.M.; Yu, X.J.; Walker, D.H.; McBride, J.W.; Kyrpides, N.C.

    2005-09-01

    Ehrlichia canis, a small obligately intracellular, tick-transmitted, gram-negative, a-proteobacterium is the primary etiologic agent of globally distributed canine monocytic ehrlichiosis. Complete genome sequencing revealed that the E. canis genome consists of a single circular chromosome of 1,315,030 bp predicted to encode 925 proteins, 40 stable RNA species, and 17 putative pseudogenes, and a substantial proportion of non-coding sequence (27 percent). Interesting genome features include a large set of proteins with transmembrane helices and/or signal sequences, and a unique serine-threonine bias associated with the potential for O-glycosylation that was prominent in proteins associated with pathogen-host interactions. Furthermore, two paralogous protein families associated with immune evasion were identified, one of which contains poly G:C tracts, suggesting that they may play a role in phase variation and facilitation of persistent infections. Proteins associated with pathogen-host interactions were identified including a small group of proteins (12) with tandem repeats and another with eukaryotic-like ankyrin domains (7).

  18. The Genome Sequence of Rickettsia felis Identifies the First Putative Conjugative Plasmid in an Obligate Intracellular Parasite

    PubMed Central

    2005-01-01

    We sequenced the genome of Rickettsia felis, a flea-associated obligate intracellular ?-proteobacterium causing spotted fever in humans. Besides a circular chromosome of 1,485,148 bp, R. felis exhibits the first putative conjugative plasmid identified among obligate intracellular bacteria. This plasmid is found in a short (39,263 bp) and a long (62,829 bp) form. R. felis contrasts with previously sequenced Rickettsia in terms of many other features, including a number of transposases, several chromosomal toxin–antitoxin genes, many more spoT genes, and a very large number of ankyrin- and tetratricopeptide-motif-containing genes. Host-invasion-related genes for patatin and RickA were found. Several phenotypes predicted from genome analysis were experimentally tested: conjugative pili and mating were observed, as well as ?-lactamase activity, actin-polymerization-driven mobility, and hemolytic properties. Our study demonstrates that complete genome sequencing is the fastest approach to reveal phenotypic characters of recently cultured obligate intracellular bacteria. PMID:15984913

  19. Motor-driven intracellular transport powers bacterial gliding motility

    PubMed Central

    Sun, Mingzhai; Wartel, Morgane; Cascales, Eric; Shaevitz, Joshua W.; Mignot, Tâm

    2011-01-01

    Protein-directed intracellular transport has not been observed in bacteria despite the existence of dynamic protein localization and a complex cytoskeleton. However, protein trafficking has clear potential uses for important cellular processes such as growth, development, chromosome segregation, and motility. Conflicting models have been proposed to explain Myxococcus xanthus motility on solid surfaces, some favoring secretion engines at the rear of cells and others evoking an unknown class of molecular motors distributed along the cell body. Through a combination of fluorescence imaging, force microscopy, and genetic manipulation, we show that membrane-bound cytoplasmic complexes consisting of motor and regulatory proteins are directionally transported down the axis of a cell at constant velocity. This intracellular motion is transmitted to the exterior of the cell and converted to traction forces on the substrate. Thus, this study demonstrates the existence of a conserved class of processive intracellular motors in bacteria and shows how these motors have been adapted to produce cell motility. PMID:21482768

  20. Draft genome sequences for the obligate bacterial predators Bacteriovorax spp. of four phylogenetic clusters

    PubMed Central

    2015-01-01

    Bacteriovorax is the halophilic genus of the obligate bacterial predators, Bdellovibrio and like organisms. The predators are known for their unique biphasic life style in which they search for and attack their prey in the free living phase; penetrate, grow, multiply and lyse the prey in the intraperiplasmic phase. Bacteriovorax isolates representing four phylogenetic clusters were selected for genomic sequencing. Only one type strain genome has been published so far from the genus Bacteriovorax. We report the genomes from non-type strains isolated from aquatic environments. Here we describe and compare the genomic features of the four strains, together with the classification and annotation. PMID:26203326

  1. Intracellular Bacterial Communities: A Potential Etiology for Chronic Lower Urinary Tract Symptoms.

    PubMed

    Scott, Victoria C S; Haake, David A; Churchill, Bernard M; Justice, Sheryl S; Kim, Ja-Hong

    2015-09-01

    Patients with persistent lower urinary tract symptoms and negative urine cultures are often difficult to treat. Infection may go undetected in these patients because the concentrations of bacteria in their urine are beneath the threshold of standard urine culture techniques. Empiric treatment may result in temporary relief, followed by recurrent symptoms. Occult and recurrent urinary tract infection may be due to both invasion of the bladder wall by uropathogenic Escherichia coli and the formation of biofilm-like intracellular bacterial communities. This review examines emerging evidence for a role of intracellular bacterial communities in human infection. PMID:26189137

  2. Directed antigen delivery as a vaccine strategy for an intracellular bacterial pathogen

    NASA Astrophysics Data System (ADS)

    Bouwer, H. G. Archie; Alberti-Segui, Christine; Montfort, Megan J.; Berkowitz, Nathan D.; Higgins, Darren E.

    2006-03-01

    We have developed a vaccine strategy for generating an attenuated strain of an intracellular bacterial pathogen that, after uptake by professional antigen-presenting cells, does not replicate intracellularly and is readily killed. However, after degradation of the vaccine strain within the phagolysosome, target antigens are released into the cytosol for endogenous processing and presentation for stimulation of CD8+ effector T cells. Applying this strategy to the model intracellular pathogen Listeria monocytogenes, we show that an intracellular replication-deficient vaccine strain is cleared rapidly in normal and immunocompromised animals, yet antigen-specific CD8+ effector T cells are stimulated after immunization. Furthermore, animals immunized with the intracellular replication-deficient vaccine strain are resistant to lethal challenge with a virulent WT strain of L. monocytogenes. These studies suggest a general strategy for developing safe and effective, attenuated intracellular replication-deficient vaccine strains for stimulation of protective immune responses against intracellular bacterial pathogens. CD8+ T cell | replication-deficient | Listeria monocytogenes

  3. Host-Directed Antimicrobial Drugs with Broad-Spectrum Efficacy against Intracellular Bacterial Pathogens

    PubMed Central

    Czy?, Daniel M.; Potluri, Lakshmi-Prasad; Jain-Gupta, Neeta; Riley, Sean P.; Martinez, Juan J.; Steck, Theodore L.; Crosson, Sean; Gabay, Joëlle E.

    2014-01-01

    ABSTRACT We sought a new approach to treating infections by intracellular bacteria, namely, by altering host cell functions that support their growth. We screened a library of 640 Food and Drug Administration (FDA)-approved compounds for agents that render THP-1 cells resistant to infection by four intracellular pathogens. We identified numerous drugs that are not antibiotics but were highly effective in inhibiting intracellular bacterial growth with limited toxicity to host cells. These compounds are likely to target three kinds of host functions: (i) G protein-coupled receptors, (ii) intracellular calcium signals, and (iii) membrane cholesterol distribution. The compounds that targeted G protein receptor signaling and calcium fluxes broadly inhibited Coxiella burnetii, Legionella pneumophila, Brucella abortus, and Rickettsia conorii, while those directed against cholesterol traffic strongly attenuated the intracellular growth of C. burnetii and L. pneumophila. These pathways probably support intracellular pathogen growth so that drugs that perturb them may be therapeutic candidates. Combining host- and pathogen-directed treatments is a strategy to decrease the emergence of drug-resistant intracellular bacterial pathogens. PMID:25073644

  4. Host-derived glucose and its transporter in the obligate intracellular pathogen Toxoplasma gondii are dispensable by glutaminolysis

    PubMed Central

    Blume, Martin; Rodriguez-Contreras, Dayana; Landfear, Scott; Fleige, Tobias; Soldati-Favre, Dominique; Lucius, Richard; Gupta, Nishith

    2009-01-01

    Toxoplasma gondii, as an obligate intracellular and promiscuous pathogen of mammalian cells, utilizes host sugars for energy and to generate glycoconjugates that are important to its survival and virulence. Here, we report that T. gondii glucose transporter (TgGT1) is proficient in transporting mannose, galactose, and fructose besides glucose, and serves as a major hexose transporter at its plasma membrane. Toxoplasma harbors 3 additional putative sugar transporters (TgST1–3), of which TgST2 is expressed at its surface, whereas TgST1 and TgST3 are intracellular. Surprisingly, TgGT1 and TgST2 are nonessential to the parasite as their ablations inflict only a 30% or no defect in its intracellular growth, respectively. Indeed, Toxoplasma can also tolerate the deletion of both genes while incurring no further growth phenotype. Unlike ?tgst2, the modest impairment in ?tggt1 and ?tggt1/?tgst2 mutants is because of a minor delay in their intracellular replication, which is a direct consequence of the abolished import of glucose. The ?tggt1 displays an attenuated motility in defined minimal media that is rescued by glutamine. TgGT1-complemented parasites show an entirely restored growth, motility, and sugar import. The lack of exogenous glucose in ?tggt1 culture fails to accentuate its intrinsic growth defect and prompts it to procure glutamine to sustain its metabolism. Unexpectedly, in vivo virulence of ?tggt1 in mice remains unaffected. Taken together, our data demonstrate that glucose is nonessential for T. gondii tachyzoites, underscore glutamine is a complement substrate, and provide a basis for understanding the adaptation of T. gondii to diverse host cells. PMID:19617561

  5. Directed antigen delivery as a vaccine strategy for an intracellular bacterial pathogen

    E-print Network

    Higgins, Darren

    Directed antigen delivery as a vaccine strategy for an intracellular bacterial pathogen H. G for review October 27, 2005) We have developed a vaccine strategy for generating an attenu- ated strain intra- cellularly and is readily killed. However, after degradation of the vaccine strain within

  6. An efficient system for intracellular delivery of beta-lactam antibiotics to overcome bacterial resistance

    PubMed Central

    Abed, Nadia; Saïd-Hassane, Fatouma; Zouhiri, Fatima; Mougin, Julie; Nicolas, Valérie; Desmaële, Didier; Gref, Ruxandra; Couvreur, Patrick

    2015-01-01

    The “Golden era” of antibiotics is definitely an old story and this is especially true for intracellular bacterial infections. The poor intracellular bioavailability of antibiotics reduces the efficency of many treatments and thereby promotes resistances. Therefore, the development of nanodevices coupled with antibiotics that are capable of targeting and releasing the drug into the infected-cells appears to be a promising solution to circumvent these complications. Here, we took advantage of two natural terpenes (farnesyl and geranyl) to design nanodevices for an efficient intracellular delivery of penicillin G. The covalent linkage between the terpene moieties and the antibiotic leads to formation of prodrugs that self-assemble to form nanoparticles with a high drug payload between 55–63%. Futhermore, the addition of an environmentally-sensitive bond between the antibiotic and the terpene led to an efficient antibacterial activity against the intracellular pathogen Staphylococcus aureus with reduced intracellular replication of about 99.9% compared to untreated infected cells. Using HPLC analysis, we demonstrated and quantified the intracellular release of PenG when this sensitive-bond (SB) was present on the prodrug, showing the success of this technology to deliver antibiotics directly into cells. PMID:26311631

  7. Search for microRNAs expressed by intracellular bacterial pathogens in infected mammalian cells.

    PubMed

    Furuse, Yuki; Finethy, Ryan; Saka, Hector A; Xet-Mull, Ana M; Sisk, Dana M; Smith, Kristen L Jurcic; Lee, Sunhee; Coers, Jörn; Valdivia, Raphael H; Tobin, David M; Cullen, Bryan R

    2014-01-01

    MicroRNAs are expressed by all multicellular organisms and play a critical role as post-transcriptional regulators of gene expression. Moreover, different microRNA species are known to influence the progression of a range of different diseases, including cancer and microbial infections. A number of different human viruses also encode microRNAs that can attenuate cellular innate immune responses and promote viral replication, and a fungal pathogen that infects plants has recently been shown to express microRNAs in infected cells that repress host cell immune responses and promote fungal pathogenesis. Here, we have used deep sequencing of total expressed small RNAs, as well as small RNAs associated with the cellular RNA-induced silencing complex RISC, to search for microRNAs that are potentially expressed by intracellular bacterial pathogens and translocated into infected animal cells. In the case of Legionella and Chlamydia and the two mycobacterial species M. smegmatis and M. tuberculosis, we failed to detect any bacterial small RNAs that had the characteristics expected for authentic microRNAs, although large numbers of small RNAs of bacterial origin could be recovered. However, a third mycobacterial species, M. marinum, did express an ? 23-nt small RNA that was bound by RISC and derived from an RNA stem-loop with the characteristics expected for a pre-microRNA. While intracellular expression of this candidate bacterial microRNA was too low to effectively repress target mRNA species in infected cultured cells in vitro, artificial overexpression of this potential bacterial pre-microRNA did result in the efficient repression of a target mRNA. This bacterial small RNA therefore represents the first candidate microRNA of bacterial origin. PMID:25184567

  8. Characterization of an obligate intracellular bacterium in the midgut epithelium of the bulrush bug Chilacis typhae (Heteroptera, Lygaeidae, Artheneinae).

    PubMed

    Kuechler, Stefan Martin; Dettner, Konrad; Kehl, Siegfried

    2011-05-01

    Many members of the suborder Heteroptera have symbiotic bacteria, which are usually found extracellularly in specific sacs or tubular outgrowths of the midgut or intracellularly in mycetomes. In this study, we describe the second molecular characterization of a symbiotic bacterium in a monophagous, seed-sucking stink bug of the family Lygaeidae (sensu stricto). Chilacis typhae possesses at the end of the first section of the midgut a structure which is composed of circularly arranged, strongly enlarged midgut epithelial cells. It is filled with an intracellular endosymbiont. This "mycetocytic belt" might represent an evolutionarily intermediate stage of the usual symbiotic structures found in stink bugs. Phylogenetic analysis based on the 16S rRNA and the groEL genes showed that the bacterium belongs to the Gammaproteobacteria, and it revealed a phylogenetic relationship with a secondary bacterial endosymbiont of Cimex lectularius and free-living plant pathogens such as Pectobacterium and Dickeya. The distribution and ultrastructure of the rod-shaped Chilacis endosymbiont were studied in adults and nymph stages using fluorescence in situ hybridization (FISH) and electron microscopy. The detection of symbionts at the anterior poles of developing eggs indicates that endosymbionts are transmitted vertically. A new genus and species name, "Candidatus Rohrkolberia cinguli," is proposed for this newly characterized clade of symbiotic bacteria. PMID:21378044

  9. ORIGINAL ARTICLE Detection of a novel intracellular microbiome hosted

    E-print Network

    ORIGINAL ARTICLE Detection of a novel intracellular microbiome hosted in arbuscular mycorrhizal) are important members of the plant microbiome. They are obligate biotrophs that colonize the roots of most land time that fungi support an intracellular bacterial microbiome, in which distinct types of endobacteria

  10. Infected Dendritic Cells Facilitate Systemic Dissemination and Transplacental Passage of the Obligate Intracellular Parasite Neospora caninum in Mice

    PubMed Central

    Collantes-Fernandez, Esther; Arrighi, Romanico B. G.; Álvarez-García, Gema; Weidner, Jessica M.; Regidor-Cerrillo, Javier; Boothroyd, John C.; Ortega-Mora, Luis M.; Barragan, Antonio

    2012-01-01

    The obligate intracellular parasite Neospora caninum disseminates across the placenta and the blood-brain barrier, to reach sites where it causes severe pathology or establishes chronic persistent infections. The mechanisms used by N. caninum to breach restrictive biological barriers remain elusive. To examine the cellular basis of these processes, migration of different N. caninum isolates (Nc-1, Nc-Liverpool, Nc-SweB1 and the Spanish isolates: Nc-Spain 3H, Nc-Spain 4H, Nc-Spain 6, Nc-Spain 7 and Nc-Spain 9) was studied in an in vitro model based on a placental trophoblast-derived BeWo cell line. Here, we describe that infection of dendritic cells (DC) by N. caninum tachyzoites potentiated translocation of parasites across polarized cellular monolayers. In addition, powered by the parasite's own gliding motility, extracellular N. caninum tachyzoites were able to transmigrate across cellular monolayers. Altogether, the presented data provides evidence of two putative complementary pathways utilized by N. caninum, in an isolate-specific fashion, for passage of restrictive cellular barriers. Interestingly, adoptive transfer of tachyzoite-infected DC in mice resulted in increased parasitic loads in various organs, e.g. the central nervous system, compared to infections with free parasites. Inoculation of pregnant mice with infected DC resulted in an accentuated vertical transmission to the offspring with increased parasitic loads and neonatal mortality. These findings reveal that N. caninum exploits the natural cell trafficking pathways in the host to cross cellular barriers and disseminate to deep tissues. The findings are indicative of conserved dissemination strategies among coccidian apicomplexan parasites. PMID:22403627

  11. The Omentum Is a Site of Protective IgM Production during Intracellular Bacterial Infection

    PubMed Central

    Jones, Derek D.; Racine, Rachael; Wittmer, Susan T.; Harston, Louise; Papillion, Amber M.; Dishaw, Lisa M.; Randall, Troy D.; Woodland, David L.

    2015-01-01

    Infection of mice with the bacterium Ehrlichia muris elicits a protective T cell-independent (TI) IgM response mediated primarily by a population of CD11c-expressing plasmablasts in the spleen. Although splenic marginal zone (MZ) B cells are considered to be important for TI responses to blood-borne pathogens, MZ B cells were not responsible for generating plasmablasts in response to Ehrlichia muris. Moreover, antigen-specific serum IgM was decreased only modestly in splenectomized mice and in mice that lacked spleen, lymph nodes, and Peyer's patches (SLP mice). Both splenectomized and SLP mice were protected against lethal ehrlichial challenge infection. Moreover, we found a high frequency of Ehrlichia-specific plasmablasts in the omentum of both conventional and SLP mice. Omental plasmablasts elicited during Ehrlichia infection lacked expression of CD138 but expressed CD11c in a manner similar to that of their splenic counterparts. Selective ablation of CD11c-expressing B cells nearly eliminated the omental Ehrlichia-specific plasmablasts and reduced antigen-specific serum IgM, identifying the omental B cells as a source of IgM production in the SLP mice. Generation of the omental plasmablasts was route dependent, as they were detected following peritoneal infection but not following intravenous infection. Our data identify the omentum as an important auxiliary site of IgM production during intracellular bacterial infection. PMID:25776744

  12. The omentum is a site of protective IgM production during intracellular bacterial infection.

    PubMed

    Jones, Derek D; Racine, Rachael; Wittmer, Susan T; Harston, Louise; Papillion, Amber M; Dishaw, Lisa M; Randall, Troy D; Woodland, David L; Winslow, Gary M

    2015-05-01

    Infection of mice with the bacterium Ehrlichia muris elicits a protective T cell-independent (TI) IgM response mediated primarily by a population of CD11c-expressing plasmablasts in the spleen. Although splenic marginal zone (MZ) B cells are considered to be important for TI responses to blood-borne pathogens, MZ B cells were not responsible for generating plasmablasts in response to Ehrlichia muris. Moreover, antigen-specific serum IgM was decreased only modestly in splenectomized mice and in mice that lacked spleen, lymph nodes, and Peyer's patches (SLP mice). Both splenectomized and SLP mice were protected against lethal ehrlichial challenge infection. Moreover, we found a high frequency of Ehrlichia-specific plasmablasts in the omentum of both conventional and SLP mice. Omental plasmablasts elicited during Ehrlichia infection lacked expression of CD138 but expressed CD11c in a manner similar to that of their splenic counterparts. Selective ablation of CD11c-expressing B cells nearly eliminated the omental Ehrlichia-specific plasmablasts and reduced antigen-specific serum IgM, identifying the omental B cells as a source of IgM production in the SLP mice. Generation of the omental plasmablasts was route dependent, as they were detected following peritoneal infection but not following intravenous infection. Our data identify the omentum as an important auxiliary site of IgM production during intracellular bacterial infection. PMID:25776744

  13. Effect of intracellular pH on the torque-speed relationship of bacterial proton-driven flagellar motor.

    PubMed

    Nakamura, Shuichi; Kami-ike, Nobunori; Yokota, Jun-ichi P; Kudo, Seishi; Minamino, Tohru; Namba, Keiichi

    2009-02-20

    Bacterial flagella responsible for motility are driven by rotary motors powered by the electrochemical potential difference of specific ions across the cytoplasmic membrane. The stator of proton-driven flagellar motor converts proton influx into mechanical work. However, the energy conversion mechanism remains unclear. Here, we show that the motor is sensitive to intracellular proton concentration for high-speed rotation at low load, which was considerably impaired by lowering intracellular pH, while zero-speed torque was not affected. The change in extracellular pH did not show any effect. These results suggest that a high intracellular proton concentration decreases the rate of proton translocation and therefore that of the mechanochemical reaction cycle of the motor but not the actual torque generation step within the cycle by the stator-rotor interactions. PMID:19133273

  14. Activation of Pattern Recognition Receptors Upregulates Metallothioneins, Thereby Increasing Intracellular Accumulation of Zinc, Autophagy, and Bacterial Clearance by Macrophages

    PubMed Central

    Lahiri, Amit; Abraham, Clara

    2014-01-01

    Background & Aims Continuous stimulation of pattern recognition receptors (PRRs), including nucleotide-binding oligomerization domain-2 (NOD2) (variants in NOD2 have been associated with Crohn's disease), alters the phenotype of myeloid-derived cells, reducing production of inflammatory cytokines and increasing clearance of microbes. We investigated the mechanisms by which microbial clearance increases in macrophages under these conditions. METHODS Monocytes were purified from human peripheral blood mononuclear cells and differentiated to monocyte-derived macrophages (MDMs). We also isolated human intestinal macrophages. Bacterial clearance by MDMs was assessed in gentamicin protection assays. Effects of intracellular zinc and autophagy were measured by flow cytometry, immunoblot, reverse transcription PCR, and microscopy experiments. Small interfering RNAs were used to knock down specific proteins in MDMs. NOD2–/– and C57BL/6J mice, maintained in a specific pathogen-free facility, were given antibiotics, muramyl dipeptide (to stimulate NOD2), or dextran sodium sulfate; intestinal lamina propria cells were collected and analyzed. RESULTS Chronic stimulation of human MDMs through NOD2 upregulated the expression of multiple genes encoding metallothioneins, which bind and regulate levels of intracellular zinc. Intestinal myeloid-derived cells are continually stimulated through PRRs; metallothionein expression was upregulated in human and mouse intestinal myeloid-derived cells. Continuous stimulation of NOD2 increased levels of intracellular zinc, thereby increasing autophagy and bacterial clearance. The metal-regulatory transcription factor-1 (MTF-1) was required for regulation of metallothionein genes in human MDMs. Knockdown of MTF-1 did not affect baseline clearance of bacteria by MDMs. However, the increase in intracellular zinc, autophagy, and bacterial clearance observed with continuous NOD2 stimulation was impaired in MDMs upon MTF-1 knockdown. Addition of zinc or induction of autophagy restored bacterial clearance to MDMs following metallothionein knockdown. NOD2 synergized with the PRRs TLR5 and TLR9 to increase the effects of metallothioneins in MDMs. In mice, the intestinal microbiota contributed to the regulation in expression of metallothioneins, levels of zinc, autophagy, and bacterial clearance by intestinal macrophages. CONCLUSIONS In studies of human MDMs and in mice, continuous stimulation of PRRs induces expression of metallothioneins. This leads to increased levels of intracellular zinc and enhanced clearance of bacteria via autophagy in macrophages. PMID:24960189

  15. Metabolic Cooperation of Glucose and Glutamine Is Essential for the Lytic Cycle of Obligate Intracellular Parasite Toxoplasma gondii.

    PubMed

    Nitzsche, Richard; Zagoriy, Vyacheslav; Lucius, Richard; Gupta, Nishith

    2016-01-01

    Toxoplasma gondii is a widespread protozoan parasite infecting nearly all warm-blooded organisms. Asexual reproduction of the parasite within its host cells is achieved by consecutive lytic cycles, which necessitates biogenesis of significant energy and biomass. Here we show that glucose and glutamine are the two major physiologically important nutrients used for the synthesis of macromolecules (ATP, nucleic acid, proteins, and lipids) in T. gondii, and either of them is sufficient to ensure the parasite survival. The parasite can counteract genetic ablation of its glucose transporter by increasing the flux of glutamine-derived carbon through the tricarboxylic acid cycle and by concurrently activating gluconeogenesis, which guarantee a continued biogenesis of ATP and biomass for host-cell invasion and parasite replication, respectively. In accord, a pharmacological inhibition of glutaminolysis or oxidative phosphorylation arrests the lytic cycle of the glycolysis-deficient mutant, which is primarily a consequence of impaired invasion due to depletion of ATP. Unexpectedly, however, intracellular parasites continue to proliferate, albeit slower, notwithstanding a simultaneous deprivation of glucose and glutamine. A growth defect in the glycolysis-impaired mutant is caused by a compromised synthesis of lipids, which cannot be counterbalanced by glutamine but can be restored by acetate. Consistently, supplementation of parasite cultures with exogenous acetate can amend the lytic cycle of the glucose transport mutant. Such plasticity in the parasite's carbon flux enables a growth-and-survival trade-off in assorted nutrient milieus, which may underlie the promiscuous survival of T. gondii tachyzoites in diverse host cells. Our results also indicate a convergence of parasite metabolism with cancer cells. PMID:26518878

  16. Activator of G-Protein Signaling 3-Induced Lysosomal Biogenesis Limits Macrophage Intracellular Bacterial Infection.

    PubMed

    Vural, Ali; Al-Khodor, Souhaila; Cheung, Gordon Y C; Shi, Chong-Shan; Srinivasan, Lalitha; McQuiston, Travis J; Hwang, Il-Young; Yeh, Anthony J; Blumer, Joe B; Briken, Volker; Williamson, Peter R; Otto, Michael; Fraser, Iain D C; Kehrl, John H

    2016-01-15

    Many intracellular pathogens cause disease by subverting macrophage innate immune defense mechanisms. Intracellular pathogens actively avoid delivery to or directly target lysosomes, the major intracellular degradative organelle. In this article, we demonstrate that activator of G-protein signaling 3 (AGS3), an LPS-inducible protein in macrophages, affects both lysosomal biogenesis and activity. AGS3 binds the Gi family of G proteins via its G-protein regulatory (GoLoco) motif, stabilizing the G? subunit in its GDP-bound conformation. Elevated AGS3 levels in macrophages limited the activity of the mammalian target of rapamycin pathway, a sensor of cellular nutritional status. This triggered the nuclear translocation of transcription factor EB, a known activator of lysosomal gene transcription. In contrast, AGS3-deficient macrophages had increased mammalian target of rapamycin activity, reduced transcription factor EB activity, and a lower lysosomal mass. High levels of AGS3 in macrophages enhanced their resistance to infection by Burkholderia cenocepacia J2315, Mycobacterium tuberculosis, and methicillin-resistant Staphylococcus aureus, whereas AGS3-deficient macrophages were more susceptible. We conclude that LPS priming increases AGS3 levels, which enhances lysosomal function and increases the capacity of macrophages to eliminate intracellular pathogens. PMID:26667172

  17. Assay Development for Image-Based Quantification of Intracellular Bacterial Replication and Analysis of the Innate Immune Response to Infection.

    PubMed

    Miller, Alexandra H; Vayttaden, Sharat J; Al-Khodor, Souhaila; Fraser, Iain D C

    2015-11-01

    Severe bacterial infection can lead to inflammation, host tissue damage, and ultimately disseminated septic shock. The mammalian innate immune system responds to microbial infection through the detection of invariant pathogen-associated molecular patterns (PAMPs) by a range of pattern recognition receptors (PRRs) expressed by the host cell. A successful immune response involves tightly coordinated signaling from these receptors, leading to a robust transcriptional response producing cytokines and antimicrobial effectors. While the PRR-expressing phagocytes of the host innate immune system function to contain and degrade internalized bacteria through pathways such as selective autophagy, pathogenic bacteria may subvert this process to replicate in the host cell. We describe the development of imaging assays to investigate these host-pathogen interactions through gene perturbation screens, which could lead to the identification of novel effectors of the host response to bacterial infection. We identify markers of coordinated initial signaling in macrophages challenged with ligands to PRRs of the toll-like receptor (TLR) family and compare this response to that induced by intact bacteria of the Burkholderia cenocepacia complex (Bcc), an opportunistic pathogen that causes life-threatening infections in patients with cystic fibrosis and chronic granulomatous disease. Bcc has been shown to escape the endocytic pathway, activate selective autophagy, and replicate within human macrophages. We demonstrate robust image-based quantification of multiple stages of Bcc infection of macrophages: ubiquitin tagging of cytosolic bacteria, recruitment of selective autophagy effector proteins, and intracellular bacterial replication, and we show perturbation of bacterial replication using drug treatment or siRNA-based gene knockdown. The described panel of imaging assays can be extended to other bacterial infections and pathogenic ligand combinations where high-content siRNA screening could provide significant new insight into regulation of the innate immune response to infection. PMID:26505731

  18. Specific Behavior of Intracellular Streptococcus pyogenes That Has Undergone Autophagic Degradation Is Associated with Bacterial Streptolysin O and Host Small G Proteins Rab5 and Rab7*

    PubMed Central

    Sakurai, Atsuo; Maruyama, Fumito; Funao, Junko; Nozawa, Takashi; Aikawa, Chihiro; Okahashi, Nobuo; Shintani, Seikou; Hamada, Shigeyuki; Ooshima, Takashi; Nakagawa, Ichiro

    2010-01-01

    Streptococcus pyogenes (group A streptococcus (GAS)) is a pathogen that invades non-phagocytic host cells, and causes a variety of acute infections such as pharyngitis. Our group previously reported that intracellular GAS is effectively degraded by the host-cell autophagic machinery, and that a cholesterol-dependent cytolysin, streptolysin O (SLO), is associated with bacterial escape from endosomes in epithelial cells. However, the details of both the intracellular behavior of GAS and the process leading to its autophagic degradation remain unknown. In this study, we found that two host small G proteins, Rab5 and Rab7, were associated with the pathway of autophagosome formation and the fate of intracellular GAS. Rab5 was involved in bacterial invasion and endosome fusion. Rab7 was clearly multifunctional, with roles in bacterial invasion, endosome maturation, and autophagosome formation. In addition, this study showed that the bacterial cytolysin SLO supported the escape of GAS into the cytoplasm from endosomes, and surprisingly, a SLO-deficient mutant of GAS was viable longer than the wild-type strain although it failed to escape the endosomes. This intracellular behavior of GAS is unique and distinct from that of other types of bacterial invaders. Our results provide a new picture of GAS infection and host-cell responses in epithelial cells. PMID:20472552

  19. Specific behavior of intracellular Streptococcus pyogenes that has undergone autophagic degradation is associated with bacterial streptolysin O and host small G proteins Rab5 and Rab7.

    PubMed

    Sakurai, Atsuo; Maruyama, Fumito; Funao, Junko; Nozawa, Takashi; Aikawa, Chihiro; Okahashi, Nobuo; Shintani, Seikou; Hamada, Shigeyuki; Ooshima, Takashi; Nakagawa, Ichiro

    2010-07-16

    Streptococcus pyogenes (group A streptococcus (GAS)) is a pathogen that invades non-phagocytic host cells, and causes a variety of acute infections such as pharyngitis. Our group previously reported that intracellular GAS is effectively degraded by the host-cell autophagic machinery, and that a cholesterol-dependent cytolysin, streptolysin O (SLO), is associated with bacterial escape from endosomes in epithelial cells. However, the details of both the intracellular behavior of GAS and the process leading to its autophagic degradation remain unknown. In this study, we found that two host small G proteins, Rab5 and Rab7, were associated with the pathway of autophagosome formation and the fate of intracellular GAS. Rab5 was involved in bacterial invasion and endosome fusion. Rab7 was clearly multifunctional, with roles in bacterial invasion, endosome maturation, and autophagosome formation. In addition, this study showed that the bacterial cytolysin SLO supported the escape of GAS into the cytoplasm from endosomes, and surprisingly, a SLO-deficient mutant of GAS was viable longer than the wild-type strain although it failed to escape the endosomes. This intracellular behavior of GAS is unique and distinct from that of other types of bacterial invaders. Our results provide a new picture of GAS infection and host-cell responses in epithelial cells. PMID:20472552

  20. A Rickettsia Genome Overrun by Mobile Genetic Elements Provides Insight into the Acquisition of Genes Characteristic of an Obligate Intracellular Lifestyle

    PubMed Central

    Joardar, Vinita; Williams, Kelly P.; Driscoll, Timothy; Hostetler, Jessica B.; Nordberg, Eric; Shukla, Maulik; Walenz, Brian; Hill, Catherine A.; Nene, Vishvanath M.; Azad, Abdu F.; Sobral, Bruno W.; Caler, Elisabet

    2012-01-01

    We present the draft genome for the Rickettsia endosymbiont of Ixodes scapularis (REIS), a symbiont of the deer tick vector of Lyme disease in North America. Among Rickettsia species (Alphaproteobacteria: Rickettsiales), REIS has the largest genome sequenced to date (>2 Mb) and contains 2,309 genes across the chromosome and four plasmids (pREIS1 to pREIS4). The most remarkable finding within the REIS genome is the extraordinary proliferation of mobile genetic elements (MGEs), which contributes to a limited synteny with other Rickettsia genomes. In particular, an integrative conjugative element named RAGE (for Rickettsiales amplified genetic element), previously identified in scrub typhus rickettsiae (Orientia tsutsugamushi) genomes, is present on both the REIS chromosome and plasmids. Unlike the pseudogene-laden RAGEs of O. tsutsugamushi, REIS encodes nine conserved RAGEs that include F-like type IV secretion systems similar to that of the tra genes encoded in the Rickettsia bellii and R. massiliae genomes. An unparalleled abundance of encoded transposases (>650) relative to genome size, together with the RAGEs and other MGEs, comprise ?35% of the total genome, making REIS one of the most plastic and repetitive bacterial genomes sequenced to date. We present evidence that conserved rickettsial genes associated with an intracellular lifestyle were acquired via MGEs, especially the RAGE, through a continuum of genomic invasions. Robust phylogeny estimation suggests REIS is ancestral to the virulent spotted fever group of rickettsiae. As REIS is not known to invade vertebrate cells and has no known pathogenic effects on I. scapularis, its genome sequence provides insight on the origin of mechanisms of rickettsial pathogenicity. PMID:22056929

  1. Intracellular Bacterial Pathogens Trigger the Formation of U Small Nuclear RNA Bodies (U Bodies) through Metabolic Stress Induction.

    PubMed

    Tsalikis, Jessica; Tattoli, Ivan; Ling, Arthur; Sorbara, Matthew T; Croitoru, David O; Philpott, Dana J; Girardin, Stephen E

    2015-08-21

    Invasive bacterial pathogens induce an amino acid starvation (AAS) response in infected host cells that controls host defense in part by promoting autophagy. However, whether AAS has additional significant effects on the host response to intracellular bacteria remains poorly characterized. Here we showed that Shigella, Salmonella, and Listeria interfere with spliceosomal U snRNA maturation in the cytosol. Bacterial infection resulted in the rerouting of U snRNAs and their cytoplasmic escort, the survival motor neuron (SMN) complex, to processing bodies, thus forming U snRNA bodies (U bodies). This process likely contributes to the decline in the cytosolic levels of U snRNAs and of the SMN complex proteins SMN and DDX20 that we observed in infected cells. U body formation was triggered by membrane damage in infected cells and was associated with the induction of metabolic stresses, such as AAS or endoplasmic reticulum stress. Mechanistically, targeting of U snRNAs to U bodies was regulated by translation initiation inhibition and the ATF4/ATF3 pathway, and U bodies rapidly disappeared upon removal of the stress, suggesting that their accumulation represented an adaptive response to metabolic stress. Importantly, this process likely contributed to shape the host response to invasive bacteria because down-regulation of DDX20 expression using short hairpin RNA (shRNA) amplified ATF3- and NF-?B-dependent signaling. Together, these results identify a critical role for metabolic stress and invasive bacterial pathogens in U body formation and suggest that this process contributes to host defense. PMID:26134566

  2. Invasion of the central nervous system by intracellular bacteria.

    PubMed

    Drevets, Douglas A; Leenen, Pieter J M; Greenfield, Ronald A

    2004-04-01

    Infection of the central nervous system (CNS) is a severe and frequently fatal event during the course of many diseases caused by microbes with predominantly intracellular life cycles. Examples of these include the facultative intracellular bacteria Listeria monocytogenes, Mycobacterium tuberculosis, and Brucella and Salmonella spp. and obligate intracellular microbes of the Rickettsiaceae family and Tropheryma whipplei. Unfortunately, the mechanisms used by intracellular bacterial pathogens to enter the CNS are less well known than those used by bacterial pathogens with an extracellular life cycle. The goal of this review is to elaborate on the means by which intracellular bacterial pathogens establish infection within the CNS. This review encompasses the clinical and pathological findings that pertain to the CNS infection in humans and includes experimental data from animal models that illuminate how these microbes enter the CNS. Recent experimental data showing that L. monocytogenes can invade the CNS by more than one mechanism make it a useful model for discussing the various routes for neuroinvasion used by intracellular bacterial pathogens. PMID:15084504

  3. Applying an Inducible Expression System to Study Interference of Bacterial Virulence Factors with Intracellular Signaling.

    PubMed

    Berens, Christian; Bisle, Stephanie; Klingenbeck, Leonie; Lührmann, Anja

    2015-01-01

    The technique presented here allows one to analyze at which step a target protein, or alternatively a small molecule, interacts with the components of a signaling pathway. The method is based, on the one hand, on the inducible expression of a specific protein to initiate a signaling event at a defined and predetermined step in the selected signaling cascade. Concomitant expression, on the other hand, of the gene of interest then allows the investigator to evaluate if the activity of the expressed target protein is located upstream or downstream of the initiated signaling event, depending on the readout of the signaling pathway that is obtained. Here, the apoptotic cascade was selected as a defined signaling pathway to demonstrate protocol functionality. Pathogenic bacteria, such as Coxiella burnetii, translocate effector proteins that interfere with host cell death induction in the host cell to ensure bacterial survival in the cell and to promote their dissemination in the organism. The C. burnetii effector protein CaeB effectively inhibits host cell death after induction of apoptosis with UV-light or with staurosporine. To narrow down at which step CaeB interferes with the propagation of the apoptotic signal, selected proteins with well-characterized pro-apoptotic activity were expressed transiently in a doxycycline-inducible manner. If CaeB acts upstream of these proteins, apoptosis will proceed unhindered. If CaeB acts downstream, cell death will be inhibited. The test proteins selected were Bax, which acts at the level of the mitochondria, and caspase 3, which is the major executioner protease. CaeB interferes with cell death induced by Bax expression, but not by caspase 3 expression. CaeB, thus, interacts with the apoptotic cascade between these two proteins. PMID:26168006

  4. Carbon based nutrition of Staphylococcus aureus and the role of sugar phosphate transporters in intracellular bacterial replication 

    E-print Network

    Bell, John Alexander

    2014-06-28

    The Gram positive bacterium Staphylococcus aureus is a major cause of human disease in industrialized countries. This multifaceted pathogen is adapted to thrive in a variety of host niches, including the intracellular ...

  5. Anaplasma phagocytophilum APH_1387 is expressed throughout bacterial intracellular development and localizes to the pathogen-occupied vacuolar membrane.

    PubMed

    Huang, Bernice; Troese, Matthew J; Ye, Shaojing; Sims, Jonathan T; Galloway, Nathan L; Borjesson, Dori L; Carlyon, Jason A

    2010-05-01

    Obligate vacuolar pathogens produce proteins that localize to the host cell-derived membranes of the vacuoles in which they reside, yielding unique organelles that are optimally suited for pathogen survival. Anaplasma phagocytophilum is an obligate vacuolar bacterium that infects neutrophils and causes the emerging and potentially fatal disease human granulocytic anaplasmosis. Here we identified APH_1387 as the first A. phagocytophilum-derived protein that associates with the A. phagocytophilum-occupied vacuolar membrane (AVM). APH_1387, also referred to as P100, is a 61.4-kDa acidic protein that migrates with an apparent molecular weight of 115 kDa on SDS-PAGE gels. It carries 3 tandem direct repeats that comprise 58% of the protein. Each APH_1387 repeat carries a bilobed hydrophobic alpha-helix domain, which is a structural characteristic that is consistent with the structure of chlamydia-derived proteins that traverse inclusion membranes. APH_1387 is not detectable on the surfaces of A. phagocytophilum dense core organisms bound at the HL-60 cell surface, but abundant APH_1387 is detected on the surfaces of intravacuolar reticulate cell and dense core organisms. APH_1387 accumulates on the AVM throughout infection. It associates with the AVM in human HL-60, THP-1, and HMEC-1 cells and tick ISE6 cells. APH_1387 is expressed and localizes to the AVM in neutrophils recovered from A. phagocytophilum-infected mice. This paper presents the first direct evidence that A. phagocytophilum actively modifies its host cell-derived vacuole. PMID:20212090

  6. Micro-RNA 155 is required for optimal CD8+ T cell responses to acute viral and intracellular bacterial challenges.

    PubMed

    Lind, Evan F; Elford, Alisha R; Ohashi, Pamela S

    2013-02-01

    Recent studies have begun to define the role of micro-RNAs in regulating the immune response. Micro-RNA155 (mir-155) has been shown to play a role in germinal center formation, T cell inflammation, and regulatory T cell development. In this study, we evaluated the role of mir-155 in cytotoxic T cell function. We report in this study that mice lacking mir-155 have impaired CD8(+) T cell responses to infections with lymphocytic choriomeningitis virus and the intracellular bacteria Listeria monocytogenes. We show by a series of adoptive transfer studies that the impaired CD8(+) T cell response to L. monocytogenes is T cell intrinsic. In addition, we observed that CD8(+) T cells lacking mir-155 have impaired activation of the prosurvival Akt pathway after TCR cross-linking. These data suggest that mir-155 may be a good target for therapies aimed at modulating immune responses. PMID:23275599

  7. Characterization of a lipopolysaccharide-targeted monoclonal antibody and its variable fragments as candidates for prophylaxis against the obligate intracellular bacterial pathogen Coxiella burnetii.

    PubMed

    Peng, Ying; Schoenlaub, Laura; Elliott, Alexandra; Mitchell, William J; Zhang, Guoquan

    2014-11-01

    Our previous study demonstrated that treatment of Coxiella burnetii with the phase I lipopolysaccharide (PI-LPS)-targeted monoclonal antibody (MAb) 1E4 significantly inhibited C. burnetii infection in mice, suggesting that 1E4 is a protective MAb. To determine whether passive transfer of antibodies (Abs) can provide protection against C. burnetii natural infection, we examined if passive transfer of 1E4 would protect SCID mice against C. burnetii aerosol infection. The results indicated that 1E4 conferred significant protection against aerosolized C. burnetii, suggesting that 1E4 may be useful for preventing C. burnetii natural infection. To further understand the mechanisms of 1E4-mediated protection and to test the possibility of using humanized 1E4 to prevent C. burnetii infection, we examined whether the Fab fragment of 1E4 (Fab1E4), a recombinant murine single-chain variable fragment (muscFv1E4), and a humanized single-chain variable fragment (huscFv1E4) retained the ability of 1E4 to inhibit C. burnetii infection. The results indicated that Fab1E4, muscFv1E4, and huscFv1E4 were able to inhibit C. burnetii infection in mice but that their ability to inhibit C. burnetii infection was lower than that of 1E4. In addition, treatment of C. burnetii with Fab1E4, muscFv1E4, or huscFv1E4 can block C. burnetii infection of macrophages. Interestingly, treatment of C. burnetii with huscFv1E4 can significantly reduce C. burnetii infectivity in human macrophages. This report provides the first evidence to demonstrate that the humanized variable fragments of an LPS-specific MAb can neutralize C. burnetii infection and appears to be a promising step toward the potential use of a humanized MAb as emergency prophylaxis against C. burnetii exposure. PMID:25114119

  8. Characterization of a Lipopolysaccharide-Targeted Monoclonal Antibody and Its Variable Fragments as Candidates for Prophylaxis against the Obligate Intracellular Bacterial Pathogen Coxiella burnetii

    PubMed Central

    Peng, Ying; Schoenlaub, Laura; Elliott, Alexandra; Mitchell, William J.

    2014-01-01

    Our previous study demonstrated that treatment of Coxiella burnetii with the phase I lipopolysaccharide (PI-LPS)-targeted monoclonal antibody (MAb) 1E4 significantly inhibited C. burnetii infection in mice, suggesting that 1E4 is a protective MAb. To determine whether passive transfer of antibodies (Abs) can provide protection against C. burnetii natural infection, we examined if passive transfer of 1E4 would protect SCID mice against C. burnetii aerosol infection. The results indicated that 1E4 conferred significant protection against aerosolized C. burnetii, suggesting that 1E4 may be useful for preventing C. burnetii natural infection. To further understand the mechanisms of 1E4-mediated protection and to test the possibility of using humanized 1E4 to prevent C. burnetii infection, we examined whether the Fab fragment of 1E4 (Fab1E4), a recombinant murine single-chain variable fragment (muscFv1E4), and a humanized single-chain variable fragment (huscFv1E4) retained the ability of 1E4 to inhibit C. burnetii infection. The results indicated that Fab1E4, muscFv1E4, and huscFv1E4 were able to inhibit C. burnetii infection in mice but that their ability to inhibit C. burnetii infection was lower than that of 1E4. In addition, treatment of C. burnetii with Fab1E4, muscFv1E4, or huscFv1E4 can block C. burnetii infection of macrophages. Interestingly, treatment of C. burnetii with huscFv1E4 can significantly reduce C. burnetii infectivity in human macrophages. This report provides the first evidence to demonstrate that the humanized variable fragments of an LPS-specific MAb can neutralize C. burnetii infection and appears to be a promising step toward the potential use of a humanized MAb as emergency prophylaxis against C. burnetii exposure. PMID:25114119

  9. Extracellular ATP protects against sepsis through macrophage P2X7 purinergic receptors by enhancing intracellular bacterial killing.

    PubMed

    Csóka, Balázs; Németh, Zoltán H; Tör?, Gábor; Idzko, Marco; Zech, Andreas; Koscsó, Balázs; Spolarics, Zoltán; Antonioli, Luca; Cseri, Karolina; Erdélyi, Katalin; Pacher, Pál; Haskó, György

    2015-09-01

    Extracellular ATP binds to and signals through P2X7 receptors (P2X7Rs) to modulate immune function in both inflammasome-dependent and -independent manners. In this study, P2X7(-/-) mice, the pharmacological agonists ATP-magnesium salt (Mg-ATP; 100 mg/kg, EC50 ? 1.32 mM) and benzoylbenzoyl-ATP (Bz-ATP; 10 mg/kg, EC50 ? 285 ?M), and antagonist oxidized ATP (oxi-ATP; 40 mg/kg, IC50 ? 100 ?M) were used to show that P2X7R activation is crucial for the control of mortality, bacterial dissemination, and inflammation in cecal ligation and puncture-induced polymicrobial sepsis in mice. Our results with P2X7(-/-) bone marrow chimeric mice, adoptive transfer of peritoneal macrophages, and myeloid-specific P2X7(-/-) mice indicate that P2X7R signaling on macrophages is essential for the protective effect of P2X7Rs. P2X7R signaling protects through enhancing bacterial killing by macrophages, which is independent of the inflammasome. By using the connexin (Cx) channel inhibitor Gap27 (0.1 mg/kg, IC50 ? 0.25 ?M) and pannexin channel inhibitor probenecid (10 mg/kg, IC50 ? 11.7 ?M), we showed that ATP release through Cx is important for inhibiting inflammation and bacterial burden. In summary, targeting P2X7Rs provides a new opportunity for harnessing an endogenous protective immune mechanism in the treatment of sepsis. PMID:26060214

  10. Alteration of intracellular protein expressions as a key mechanism of the deterioration of bacterial denitrification caused by copper oxide nanoparticles

    NASA Astrophysics Data System (ADS)

    Su, Yinglong; Zheng, Xiong; Chen, Yinguang; Li, Mu; Liu, Kun

    2015-10-01

    The increasing production and utilization of copper oxide nanoparticles (CuO NPs) result in the releases into the environment. However, the influence of CuO NPs on bacterial denitrification, one of the most important pathways to transform nitrate to dinitrogen in environment, has seldom been studied. Here we reported that CuO NPs caused a significant alteration of key protein expressions of a model denitrifier, Paracoccus denitrificans, leading to severe inhibition to denitrification. Total nitrogen removal efficiency was decreased from 98.3% to 62.1% with the increase of CuO NPs from 0.05 to 0.25?mg/L. Cellular morphology and integrity studies indicated that nanoparticles entered the cells. The proteomic bioinformatics analysis showed that CuO NPs caused regulation of proteins involved in nitrogen metabolism, electron transfer and substance transport. The down-regulation of GtsB protein (responsible for glucose transport) decreased the production of NADH (electron donor for denitrification). Also, the expressions of key electron-transfer proteins (including NADH dehydrogenase and cytochrome) were suppressed by CuO NPs, which adversely affected electrons transfer for denitrification. Further investigation revealed that CuO NPs significantly inhibited the expressions and catalytic activities of nitrate reductase and nitrite reductase. These results provided a fundamental understanding of the negative influences of CuO NPs on bacterial denitrification.

  11. Alteration of intracellular protein expressions as a key mechanism of the deterioration of bacterial denitrification caused by copper oxide nanoparticles.

    PubMed

    Su, Yinglong; Zheng, Xiong; Chen, Yinguang; Li, Mu; Liu, Kun

    2015-01-01

    The increasing production and utilization of copper oxide nanoparticles (CuO NPs) result in the releases into the environment. However, the influence of CuO NPs on bacterial denitrification, one of the most important pathways to transform nitrate to dinitrogen in environment, has seldom been studied. Here we reported that CuO NPs caused a significant alteration of key protein expressions of a model denitrifier, Paracoccus denitrificans, leading to severe inhibition to denitrification. Total nitrogen removal efficiency was decreased from 98.3% to 62.1% with the increase of CuO NPs from 0.05 to 0.25?mg/L. Cellular morphology and integrity studies indicated that nanoparticles entered the cells. The proteomic bioinformatics analysis showed that CuO NPs caused regulation of proteins involved in nitrogen metabolism, electron transfer and substance transport. The down-regulation of GtsB protein (responsible for glucose transport) decreased the production of NADH (electron donor for denitrification). Also, the expressions of key electron-transfer proteins (including NADH dehydrogenase and cytochrome) were suppressed by CuO NPs, which adversely affected electrons transfer for denitrification. Further investigation revealed that CuO NPs significantly inhibited the expressions and catalytic activities of nitrate reductase and nitrite reductase. These results provided a fundamental understanding of the negative influences of CuO NPs on bacterial denitrification. PMID:26508362

  12. Alteration of intracellular protein expressions as a key mechanism of the deterioration of bacterial denitrification caused by copper oxide nanoparticles

    PubMed Central

    Su, Yinglong; Zheng, Xiong; Chen, Yinguang; Li, Mu; Liu, Kun

    2015-01-01

    The increasing production and utilization of copper oxide nanoparticles (CuO NPs) result in the releases into the environment. However, the influence of CuO NPs on bacterial denitrification, one of the most important pathways to transform nitrate to dinitrogen in environment, has seldom been studied. Here we reported that CuO NPs caused a significant alteration of key protein expressions of a model denitrifier, Paracoccus denitrificans, leading to severe inhibition to denitrification. Total nitrogen removal efficiency was decreased from 98.3% to 62.1% with the increase of CuO NPs from 0.05 to 0.25?mg/L. Cellular morphology and integrity studies indicated that nanoparticles entered the cells. The proteomic bioinformatics analysis showed that CuO NPs caused regulation of proteins involved in nitrogen metabolism, electron transfer and substance transport. The down-regulation of GtsB protein (responsible for glucose transport) decreased the production of NADH (electron donor for denitrification). Also, the expressions of key electron-transfer proteins (including NADH dehydrogenase and cytochrome) were suppressed by CuO NPs, which adversely affected electrons transfer for denitrification. Further investigation revealed that CuO NPs significantly inhibited the expressions and catalytic activities of nitrate reductase and nitrite reductase. These results provided a fundamental understanding of the negative influences of CuO NPs on bacterial denitrification. PMID:26508362

  13. Patho-epigenetics of Infectious Diseases Caused by Intracellular Bacteria.

    PubMed

    Niller, Hans Helmut; Minarovits, Janos

    2016-01-01

    In multicellular eukaryotes including plants, animals and humans, epigenetic reprogramming may play a role in the pathogenesis of a wide variety of diseases. Recent studies revealed that in addition to viruses, pathogenic bacteria are also capable to dysregulate the epigenetic machinery of their target cells. In this chapter we focus on epigenetic alterations induced by bacteria infecting humans. Most of them are obligate or facultative intracellular bacteria that produce either bacterial toxins and surface proteins targeting the host cell membrane, or synthesise effector proteins entering the host cell nucleus. These bacterial products typically elicit histone modifications, i.e. alter the "histone code". Bacterial pathogens are capable to induce alterations of host cell DNA methylation patterns, too. Such changes in the host cell epigenotype and gene expression pattern may hinder the antibacterial immune response and create favourable conditions for bacterial colonization, growth, or spread. Epigenetic dysregulation mediated by bacterial products may also facilitate the production of inflammatory cytokines and other inflammatory mediators affecting the epigenotype of their target cells. Such indirect epigenetic changes as well as direct interference with the epigenetic machinery of the host cells may contribute to the initiation and progression of malignant tumors associated with distinct bacterial infections. PMID:26659266

  14. Intracellular life of Coxiella burnetii in macrophages.

    PubMed

    Ghigo, Eric; Pretat, Lionel; Desnues, Benoît; Capo, Christian; Raoult, Didier; Mege, Jean-Louis

    2009-05-01

    Coxiella burnetii, the agent of Q fever, is an obligate intracellular bacterium that is considered a potential biological weapon of category B. C. burnetii survives within myeloid cells by subverting receptor-mediated phagocytosis and preventing phagosome maturation. The intracellular fate of C. burnetii also depends on the functional state of myeloid cells. This review describes the mechanisms used by C. burnetii to circumvent uptake and trafficking events, and the role of cytokines on C. burnetii survival in myeloid cells. PMID:19538264

  15. NCI & Division Obligations

    Cancer.gov

    Displays obligations for grants, contracts, training fellowships, intramural research, and management and support, including the number of grant awards, funding amounts, and percent of the total NCI budget.

  16. [Bacterial Genome Evolution in Superspecies Systems: an Approach to the Reconstruction of Symbiogenesis Processes].

    PubMed

    Provorov, N A; Tikhonovich, I A

    2015-04-01

    Bacteria form a broad spectrum of symbioses with eukaryotes. This permits reconstruction of the symbiogenesis processes providing the transformation of free-living microorganisms into cellular organelles. In ecologically (conditionally) obligate symbioses, an increase in the size and complexity of the bacterial genome structure was observed. This was associated with segregation of the regions controlling symbiosis into gene clusters, islands, and plasmids. In genetically (strictly) obligate symbioses, a reduction of "nonsymbiotic" regions of microbial genome occurs, which could begin from genes encoding metabolic and regulatory functions. It is extended towards genes encoding template processes. Conditionally obligate symbioses are characterised by the activation of horizontal gene transfer between various forms of microsymbionts, while for strictly obligate intracellular symbioses an activation of endo-symbiotic gene transfer between microsymbionts and their hosts was detected. The latter is responsible for bacterial transition from the functional (based on gene cross-regulation) to structural (based on recombination) genetic integration with hosts, which later could be followed by the complete assimilation of microbial genomes. In ?-proteobacteria this evolutionary pathway could result in the formation of cellular organelles that are deficient in their own genomes but capable of preserving proteomic and cytological traits as a result of the gene-product import synthesized in cytosol (hydrogenosomes and mitosomes). The symbiogenic evolution of cyanobacteria could result in the loss of the plasmids generated from them, while the host maintains a significant part of their genome in nuclear chromosomes. PMID:26087621

  17. Intracellular microlasers

    NASA Astrophysics Data System (ADS)

    Humar, Matjaž; Hyun Yun, Seok

    2015-09-01

    Optical microresonators, which confine light within a small cavity, are widely exploited for various applications ranging from the realization of lasers and nonlinear devices to biochemical and optomechanical sensing. Here we use microresonators and suitable optical gain materials inside biological cells to demonstrate various optical functions in vitro including lasing. We explore two distinct types of microresonator—soft and hard—that support whispering-gallery modes. Soft droplets formed by injecting oil or using natural lipid droplets support intracellular laser action. The laser spectra from oil-droplet microlasers can chart cytoplasmic internal stress (˜500?pN??m-2) and its dynamic fluctuations at a sensitivity of 20?pN??m-2 (20?Pa). In a second form, whispering-gallery modes within phagocytized polystyrene beads of different sizes enable individual tagging of thousands of cells easily and, in principle, a much larger number by multiplexing with different dyes.

  18. Modulation of innate immune signaling pathways by the intracellular pathogen Toxoplasma gondii

    E-print Network

    Rosowski, Emily E. (Emily Elizabeth)

    2013-01-01

    Toxoplasma gondii, an obligate intracellular protozoan parasite, is one of the most successful eukaryotic pathogens. It can infect virtually any warm-blooded animal, including humans, in whom it can cause serious disease. ...

  19. Obesity and obligation.

    PubMed

    Jeppsson, Sofia

    2015-03-01

    The belief that obese people ought to lose weight and keep it off is widespread, and has a profound negative impact on the lives of the obese. I argue in this paper that most obese people have no such obligation, even if obesity is bad, and caused by calorie input exceeding output. Obese people do not have an obligation to achieve long-term weight loss if this is impossible for them, is worse than the alternative, or requires such an enormous effort in relation to what stands to be gained that this option is supererogatory rather than obligatory. It is highly plausible that most obese people fall into one of these three groups. Politicians may still have obligations to fight obesity, but they ought to do so through progressive politics rather than blaming and shaming. PMID:25843121

  20. GRANDPARENTS' ENTITLEMENTS AND OBLIGATIONS

    PubMed Central

    Draper, Heather

    2013-01-01

    In this article, it is argued that grandparents' obligations originate from parental obligations (i.e from the relationship they have with their children, the parents of their grandchildren) and not from the role of grandparent per se, and any entitlements flow from the extent to which these obligations are met. The position defended is, therefore, that grandparents qua grandparents are not entitled to form or continue relationships with their grandchildren. A continuation of grandparent-grandchildren relationships may be in the interests of children, but the grandparental nature of the relationship is not decisive. What counts is the extent to which relationships children have with any adults who are not their parents are is significant to them. Sometimes, however, grandparents become parents or co-parents of their grandchildren. They then gain parental rights, and as such are as entitled, ceteris parius, as any parent to expect their relationship with the child to continue. The issue of grandparents' entitlements can come to the fore when parents separate, and grandparents are unhappy with the access they have to their grandchildren. Grandparents' obligations may become a particular issue when parents die, struggle, or fail to care for their children. This article focuses particularly on these kinds of circumstances. PMID:23718643

  1. Identification and characterization of persistent intracellular human immunodeficiency virus type 1 integrase strand transfer inhibitor activity.

    PubMed

    Koh, Yasuhiro; Haim, Hillel; Engelman, Alan

    2011-01-01

    Pharmacokinetic and pharmacodynamic considerations significantly impact infectious disease treatment options. One aspect of pharmacodynamics is the postantibiotic effect, classically defined as delayed bacterial growth after antibiotic removal. The same principle can apply to antiviral drugs. For example, significant delays in human immunodeficiency virus type 1 (HIV-1) replication can be observed after nucleoside/nucleotide reverse transcriptase inhibitor (N/NtRTI) removal from culture medium, because these prodrugs must be anabolized into active, phosphorylated forms once internalized into cells. A relatively new class of anti-HIV-1 drugs is the integrase strand transfer inhibitors (INSTIs), and the INSTIs raltegravir (RAL) and elvitegravir (EVG) were tested here alongside positive N/NtRTI controls tenofovir disoproxil fumarate (TDF) and azidothymidine (AZT), as well as the nonnucleoside reverse transcriptase inhibitor negative control nevirapine (NVP), to assess potential postantiviral effects. Transformed and primary CD4-positive cells pretreated with INSTIs significantly resisted subsequent challenge by HIV-1, revealing the following hierarchy of persistent intracellular drug strength: TDF > EVG ? AZT > RAL > NVP. A modified time-of-addition assay was moreover developed to assess residual drug activity levels. Approximately 0.8% of RAL and 2% of initial EVG and TDF 1-h pulse drug levels persisted during the acute phase of HIV-1 infection. EVG furthermore displayed significant virucidal activity. Although there is no reason to suspect obligate intracellular modification, this study nevertheless defines significant intracellular persistence of prototype INSTIs. Ongoing second-generation formulations should therefore consider the potential for significant postantiviral effects among this drug class. Combined intracellular persistence and virucidal activities suggest potential pre-exposure prophylaxis applications for EVG. PMID:21060108

  2. Cytotoxic Cells Kill Intracellular Bacteria through Granulysin-Mediated

    E-print Network

    Kirchhausen, Tomas

    granzymes into bacteria to kill diverse bacterial strains. In Escherichia coli, granzymes cleave electronCytotoxic Cells Kill Intracellular Bacteria through Granulysin-Mediated Delivery of Granzymes. What hap- pens to intracellular bacteria during this process is unclear. Human, but not rodent

  3. The olive fly endosymbiont, "Candidatus Erwinia dacicola," switches from an intracellular existence to an extracellular existence during host insect development.

    PubMed

    Estes, Anne M; Hearn, David J; Bronstein, Judith L; Pierson, Elizabeth A

    2009-11-01

    As polyphagous, holometabolous insects, tephritid fruit flies (Diptera: Tephritidae) provide a unique habitat for endosymbiotic bacteria, especially those microbes associated with the digestive system. Here we examine the endosymbiont of the olive fly [Bactrocera oleae (Rossi) (Diptera: Tephritidae)], a tephritid of great economic importance. "Candidatus Erwinia dacicola" was found in the digestive systems of all life stages of wild olive flies from the southwestern United States. PCR and microscopy demonstrated that "Ca. Erwinia dacicola" resided intracellularly in the gastric ceca of the larval midgut but extracellularly in the lumen of the foregut and ovipositor diverticulum of adult flies. "Ca. Erwinia dacicola" is one of the few nonpathogenic endosymbionts that transitions between intracellular and extracellular lifestyles during specific stages of the host's life cycle. Another unique feature of the olive fly endosymbiont is that unlike obligate endosymbionts of monophagous insects, "Ca. Erwinia dacicola" has a G+C nucleotide composition similar to those of closely related plant-pathogenic and free-living bacteria. These two characteristics of "Ca. Erwinia dacicola," the ability to transition between intracellular and extracellular lifestyles and a G+C nucleotide composition similar to those of free-living relatives, may facilitate survival in a changing environment during the development of a polyphagous, holometabolous host. We propose that insect-bacterial symbioses should be classified based on the environment that the host provides to the endosymbiont (the endosymbiont environment). PMID:19767463

  4. Phenotypic characterization and 16S rDNA identification of culturable non-obligate halophilic bacterial communities from a hypersaline lake, La Sal del Rey, in extreme South Texas (USA)

    PubMed Central

    2012-01-01

    Background La Sal del Rey ("the King's Salt") is one of several naturally-occurring salt lakes in Hidalgo County, Texas and is part of the Lower Rio Grande Valley National Wildlife Refuge. The research objective was to isolate and characterize halophilic microorganisms from La Sal del Rey. Water samples were collected from the lake and a small creek that feeds into the lake. Soil samples were collected from land adjacent to the water sample locations. Sample salinity was determined using a refractometer. Samples were diluted and cultured on a synthetic saline medium to grow halophilic bacteria. The density of halophiles was estimated by viable plate counts. A collection of isolates was selected, gram-stained, tested for catalase, and characterized using API 20E® test strips. Isolates were putatively identified by sequencing the 16S rDNA. Carbon source utilization by the microbial community from each sample site was examined using EcoPlate™ assays and the carbon utilization total activity of the community was determined. Results Results showed that salinity ranged from 4 parts per thousand (ppt) at the lake water source to 420 ppt in water samples taken just along the lake shore. The density of halophilic bacteria in water samples ranged from 1.2 × 102 - 5.2 × 103 colony forming units per ml (cfu ml-1) whereas the density in soil samples ranged from 4.0 × 105 - 2.5 × 106 colony forming units per gram (cfu g-1). In general, as salinity increased the density of the bacterial community decreased. Microbial communities from water and soil samples were able to utilize 12 - 31 carbon substrates. The greatest number of substrates utilized was by water-borne communities compared to soil-based communities, especially at lower salinities. The majority of bacteria isolated were gram-negative, catalase-positive, rods. Biochemical profiles constructed from API 20E® test strips showed that bacterial isolates from low-salinity water samples (4 ppt) showed the greatest phenotypic diversity with regards to the types and number of positive tests from the strip. Isolates taken from water samples at the highest salinity (420 ppt) tended to be less diverse and have only a limited number of positive tests. Sequencing of 16S DNA displayed the presence of members of bacterial genera Bacillus, Halomonas, Pseudomonas, Exiguobacterium and others. The genus Bacillus was most commonly identified. None of the isolates were members of the Archaea probably due to dilution of salts in the samples. Conclusions The La Sal del Rey ecosystem supports a robust and diverse bacterial community despite the high salinity of the lake and soil. However, salinity does appear to a limiting factor with regards to the density and diversity of the bacterial communities that inhabit the lake and surrounding area. PMID:22480362

  5. Ehrlichia chaffeensis Exploits Host SUMOylation Pathways To Mediate Effector-Host Interactions and Promote Intracellular Survival

    PubMed Central

    Dunphy, Paige Selvy; Luo, Tian

    2014-01-01

    Ehrlichia chaffeensis is an obligately intracellular Gram-negative bacterium that selectively infects mononuclear phagocytes. We recently reported that E. chaffeensis utilizes a type 1 secretion (T1S) system to export tandem repeat protein (TRP) effectors and demonstrated that these effectors interact with a functionally diverse array of host proteins. By way of these interactions, TRP effectors modulate host cell functions; however, the molecular basis of these interactions and their roles in ehrlichial pathobiology are not well defined. In this study, we describe the first bacterial protein posttranslational modification (PTM) by the small ubiquitin-like modifier (SUMO). The E. chaffeensis T1S effector TRP120 is conjugated to SUMO at a carboxy-terminal canonical consensus SUMO conjugation motif in vitro and in human cells. In human cells, TRP120 was selectively conjugated with SUMO2/3 isoforms. Disruption of TRP120 SUMOylation perturbed interactions with known host proteins, through predicted SUMO interaction motif-dependent and -independent mechanisms. E. chaffeensis infection did not result in dramatic changes in the global host SUMOylated protein profile, but a robust colocalization of predominately SUMO1 with ehrlichial inclusions was observed. Inhibiting the SUMO pathway with a small-molecule inhibitor had a significant impact on E. chaffeensis replication and recruitment of the TRP120-interacting protein polycomb group ring finger protein 5 (PCGF5) to the inclusion, indicating that the SUMO pathway is critical for intracellular survival. This study reveals the novel exploitation of the SUMO pathway by Ehrlichia, which facilitates effector-eukaryote interactions necessary to usurp the host and create a permissive intracellular niche. PMID:25047847

  6. Endosymbiosis In Statu Nascendi: Close Phylogenetic RelationshipBetween Obligately Endosymbiotic and Obligately Free-LivingPolynucleobacter Strains (Betaproteobacteria)

    SciTech Connect

    Vannini, Claudia; Pockl, Matthias; Petroni, Giulio; Wu, Qinglong; Lang, Elke; Stackebrandt, Erko; Schrallhammer, Martina; Richardson, PaulM.; Hahn, Martin W.

    2006-07-21

    Bacterial strains affiliated to the phylogenetically shallowsubcluster C (PnecC) of the 28 Polynucleobacter cluster, which ischaracterized by a minimal 16S rRNA gene sequence similarity of approx.98.5 percent, have been reported to occur as obligate endosymbionts of 30ciliates (Euplotes spp.), as well as to occur as free-living cells in thepelagic zone of freshwater habitats. We investigated if these two groupsof closely related bacteria represent 32 strains fundamentally differingin lifestyle, or if they simply represent different stages of afacultative endosymbiotic lifestyle. The phylogenetic analysis of 16SrRNA gene and 16S34 23S ITS sequences of five endosymbiont strains fromtwo different Euplotes species and 40 pure culture strains demonstratedhost-species-specific clustering of the endosymbiont 36 sequences withinthe PnecC subcluster. The sequences of the endosymbionts showedcharacteristics indicating an obligate endosymbiotic lifestyle.Cultivation experiments 38 revealed fundamental differences inphysiological adaptations, and determination of the genome sizesindicated a slight size reduction in endosymbiotic strains. We concludethat the 40 two groups of PnecC bacteria represent obligately free-livingand obligately endosymbiotic strains, respectively, and do not representdifferent stages of the same complex lifecycle. 42 These closely relatedstrains occupy completely separated ecological niches. To our bestknowledge, this is the closest phylogenetic relationship between obligateendosymbionts and 44 obligately free-living bacteria everrevealed.

  7. Can’t Take the Heat: High Temperature Depletes Bacterial Endosymbionts of Ants

    PubMed Central

    Fan, Yongliang; Wernegreen, Jennifer J.

    2014-01-01

    Members of the ant tribe Camponotini have coevolved with Blochmannia, an obligate intracellular bacterial mutualist. This endosymbiont lives within host bacteriocyte cells that line the ant midgut, undergoes maternal transmission from host queens to offspring, and contributes to host nutrition via nitrogen recycling and nutrient biosynthesis. While elevated temperature has been shown to disrupt obligate bacterial mutualists of some insects, its impact on the ant-Blochmannia partnership is less clear. Here, we test the effect of heat on the density of Blochmannia in two related Camponotus species in the lab. Transcriptionally active Blochmannia were quantified using RT-qPCR as the ratio of Blochmannia 16S rRNA to ant host elongation factor 1-? transcripts. Our results showed that 4 weeks of heat treatment depleted active Blochmannia by >99 % in minor workers and unmated queens. However, complete elimination of Blochmannia transcripts rarely occurred, even after 16 weeks of heat treatment. Possible mechanisms of observed thermal sensitivity may include extreme AT-richness and related features of Blochmannia genomes, as well as host stress responses. Broadly, the observed depletion of an essential microbial mutualist in heat-treated ants is analogous to the loss of zooanthellae during coral bleaching. While the ecological relevance of Blochmannia’s thermal sensitivity is uncertain, our results argue that symbiont dynamics should be part of models predicting how ants and other animals will respond and adapt to a warming climate. PMID:23872930

  8. The role of autophagy in intracellular pathogen nutrient acquisition

    PubMed Central

    Steele, Shaun; Brunton, Jason; Kawula, Thomas

    2015-01-01

    Following entry into host cells intracellular pathogens must simultaneously evade innate host defense mechanisms and acquire energy and anabolic substrates from the nutrient-limited intracellular environment. Most of the potential intracellular nutrient sources are stored within complex macromolecules that are not immediately accessible by intracellular pathogens. To obtain nutrients for proliferation, intracellular pathogens must compete with the host cell for newly-imported simple nutrients or degrade host nutrient storage structures into their constituent components (fatty acids, carbohydrates, and amino acids). It is becoming increasingly evident that intracellular pathogens have evolved a wide variety of strategies to accomplish this task. One recurrent microbial strategy is to exploit host degradative processes that break down host macromolecules into simple nutrients that the microbe can use. Herein we focus on how a subset of bacterial, viral, and eukaryotic pathogens leverage the host process of autophagy to acquire nutrients that support their growth within infected cells. PMID:26106587

  9. Bacterial polyhydroxyalkanoates.

    PubMed

    Lee, S Y

    1996-01-01

    Polyhydroxyalkanoates (PHAs) are polyesters of hydroxyalkanoates (HAs) synthesized by numerous bacteria as intracellular carbon and energy storage compounds and accumulated as granules in the cytoplasm of cells. More than 80 HAs have been detected as constituents of PHAs, which allows these thermoplastic materials to have various mechanical properties resembling hard crystalline polymer or elastic rubber depending on the incorporated monomer units. Even though PHAs have been recognized as good candidates for biodegradable plastics, their high price compared with conventional plastics has limited their use in a wide range of applications. A number of bacteria including Alcaligenes eutrophus, Alcaligenes latus, Azotobacter vinelandii, methylotrophs, pseudomonads, and recombinant Escherichia coli have been employed for the production of PHAs, and the productivity of greater than 2 g PHA/L/h has been achieved. Recent advances in understanding metabolism, molecular biology, and genetics of the PHA-synthesizing bacteria and cloning of more than 20 different PHA biosynthesis genes allowed construction of various recombinant strains that were able to synthesize polyesters having different monomer units and/or to accumulate much more polymers. Also, genetically engineered plants harboring the bacterial PHA biosynthesis genes are being developed for the economical production of PHAs. Improvements in fermentation/separation technology and the development of bacterial strains or plants that more efficiently synthesize PHAs will bring the costs down to make PHAs competitive with the conventional plastics. PMID:18623547

  10. Obligately barophilic bacterium from the Mariana trench.

    PubMed Central

    Yayanos, A A; Dietz, A S; Van Boxtel, R

    1981-01-01

    An amphipod (Hirondellea gigas) was retrieved with decompression in an insulated trap from an ocean depth of 10,476 m. Bacterial isolates were obtained from the dead and cold animal by using silica gel medium incubated at 1000 bars (1 bar = 10(5) Pa) and 2 degrees C. The isolate designated MT41 was found to be obligately barophilic and did not grow at a pressure close to that of 380 bars found at average depths of the sea. The optimal generation time of about 25 hr was at 2 degrees C and 690 bars. The generation time at 2 degrees C and 1,035 bars, a pressure close to that at the depth of origin, was about 33 hr. Among the conclusions are: (i) pressure is an important determinant of zonation along the water column of the sea; (ii) some obligately barophilic bacteria survive decompressions; (iii) the pressure of optimal growth at 2 degrees C appears to be less than the pressure at the depth of origin and may be diagnostic for the depth of origin; (iv) rates of reproduction are slow yet significant and an order of magnitude greater than previously thought; and (v) much of deep-sea microbiology may have been done with spurious deep-sea organisms due to warming of samples. Images PMID:6946468

  11. Obligately barophilic bacterium from the Mariana trench.

    PubMed

    Yayanos, A A; Dietz, A S; Van Boxtel, R

    1981-08-01

    An amphipod (Hirondellea gigas) was retrieved with decompression in an insulated trap from an ocean depth of 10,476 m. Bacterial isolates were obtained from the dead and cold animal by using silica gel medium incubated at 1000 bars (1 bar = 10(5) Pa) and 2 degrees C. The isolate designated MT41 was found to be obligately barophilic and did not grow at a pressure close to that of 380 bars found at average depths of the sea. The optimal generation time of about 25 hr was at 2 degrees C and 690 bars. The generation time at 2 degrees C and 1,035 bars, a pressure close to that at the depth of origin, was about 33 hr. Among the conclusions are: (i) pressure is an important determinant of zonation along the water column of the sea; (ii) some obligately barophilic bacteria survive decompressions; (iii) the pressure of optimal growth at 2 degrees C appears to be less than the pressure at the depth of origin and may be diagnostic for the depth of origin; (iv) rates of reproduction are slow yet significant and an order of magnitude greater than previously thought; and (v) much of deep-sea microbiology may have been done with spurious deep-sea organisms due to warming of samples. PMID:6946468

  12. Bacterial DNA Sifted from the Trichoplax adhaerens (Animalia: Placozoa) Genome Project Reveals a Putative Rickettsial Endosymbiont

    PubMed Central

    Driscoll, Timothy; Gillespie, Joseph J.; Nordberg, Eric K.; Azad, Abdu F.; Sobral, Bruno W.

    2013-01-01

    Eukaryotic genome sequencing projects often yield bacterial DNA sequences, data typically considered as microbial contamination. However, these sequences may also indicate either symbiont genes or lateral gene transfer (LGT) to host genomes. These bacterial sequences can provide clues about eukaryote–microbe interactions. Here, we used the genome of the primitive animal Trichoplax adhaerens (Metazoa: Placozoa), which is known to harbor an uncharacterized Gram-negative endosymbiont, to search for the presence of bacterial DNA sequences. Bioinformatic and phylogenomic analyses of extracted data from the genome assembly (181 bacterial coding sequences [CDS]) and trace read archive (16S rDNA) revealed a dominant proteobacterial profile strongly skewed to Rickettsiales (Alphaproteobacteria) genomes. By way of phylogenetic analysis of 16S rDNA and 113 proteins conserved across proteobacterial genomes, as well as identification of 27 rickettsial signature genes, we propose a Rickettsiales endosymbiont of T. adhaerens (RETA). The majority (93%) of the identified bacterial CDS belongs to small scaffolds containing prokaryotic-like genes; however, 12 CDS were identified on large scaffolds comprised of eukaryotic-like genes, suggesting that T. adhaerens might have recently acquired bacterial genes. These putative LGTs may coincide with the placozoan’s aquatic niche and symbiosis with RETA. This work underscores the rich, and relatively untapped, resource of eukaryotic genome projects for harboring data pertinent to host–microbial interactions. The nature of unknown (or poorly characterized) bacterial species may only emerge via analysis of host genome sequencing projects, particularly if these species are resistant to cell culturing, as are many obligate intracellular microbes. Our work provides methodological insight for such an approach. PMID:23475938

  13. Antibody- and TRIM21-dependent intracellular restriction of Salmonella enterica.

    PubMed

    Rakebrandt, Nikolas; Lentes, Sabine; Neumann, Heinz; James, Leo C; Neumann-Staubitz, Petra

    2014-11-01

    TRIM21 ('tripartite motif-containing protein 21', Ro52) is a ubiquitously expressed cytosolic Fc receptor, which has a potent role in protective immunity against nonenveloped viruses. TRIM21 mediates intracellular neutralisation of antibody-coated viruses, a process called ADIN (antibody-dependent intracellular neutralisation). Our results reveal a similar mechanism to fight bacterial infections. TRIM21 is recruited to the intracellular pathogen Salmonella enterica in epithelial cells early in infection. TRIM21 does not bind directly to S. enterica, but to antibodies opsonising it. Most importantly, bacterial restriction is dependent on TRIM21 as well as on the opsonisation state of the bacteria. Finally, Salmonella and TRIM21 colocalise with the autophagosomal marker LC3, and intracellular defence is enhanced in starved cells suggesting an involvement of the autophagocytic pathway. Our data extend the protective role of TRIM21 from viruses to bacteria and thereby strengthening the general role of ADIN in cellular immunity. PMID:24920099

  14. Antibiotic uptake by cultured Atlantic cod leucocytes and effect on intracellular Francisella noatunensis subsp. noatunensis replication.

    PubMed

    Kaldestad, Marte; Haugland, Gyri T; Rønneseth, Anita; Wergeland, Heidrun I; Samuelsen, Ole Bent

    2014-02-01

    The granuloma disease caused by Francisella noatunensis subsp. noatunensis in farmed Atlantic cod has not been successfully treated by use of antibacterials, even when antibacterial resistance testing indicates a sufficient effect. The reason for this treatment failure may be the intracellular existence of the bacteria within immune cells, mainly macrophages. To investigate the effect of antibacterials on intracellular Francisella replication, we established a protocol for the detection of drugs within Atlantic cod immune cells using high-performance liquid chromatography (HPLC). When the uptake and intracellular concentrations of oxolinic acid and flumequine were analysed in isolated adherent head kidney leucocytes (HKLs) by HPLC, we found that uptake was rapid and the intracellular concentrations reflected the extracellular exposure concentrations. To investigate the effect of the antibacterial compounds on intracellular bacterial replication, adherent HKLs experimentally infected with the bacteria were analysed using flow cytometry and intracellular labelling of bacteria by specific antibodies. We found that flumequine did not inhibit intracellular bacterial replication. Unexpectedly, the results indicated that the intracellularly effiacy of the drug was reduced. The HPLC method used proved to be highly applicable for accurate determination of intracellular drug concentrations. When combined with sensitive and specific flow cytometry analyses for identification and measurement of intracellular bacterial replication, we suggest that this approach can be very valuable for the design of antibacterial treatments of intracellular pathogens. PMID:24492050

  15. Toward Intracellular Targeted Delivery of Cancer Therapeutics

    PubMed Central

    Pandya, Hetal; Debinski, Waldemar

    2013-01-01

    A number of anti-cancer drugs have their targets localized to particular intracellular compartments. These drugs reach the targets mainly through diffusion, dependent on biophysical and biochemical forces that allow cell penetration. This means that both cancer cells and normal cells will be subjected to such diffusion; hence many of these drugs, like chemotherapeutics, are potentially toxic and the concentration achieved at the site of their action is often suboptimal. The same relates to radiation that indiscriminately affects normal and diseased cells. However, nature-designed systems enable compounds present in the extracellular environment to end up inside the cell and even travel to more specific intracellular compartments. For example, viruses and bacterial toxins can more or less specifically recognize eukaryotic cells, enter these cells, and direct some protein portions to designated intracellular areas. These phenomena have led to creative thinking, such as employing viruses or bacterial toxins for cargo delivery to cells and, more specifically, to cancer cells. Proteins can be genetically engineered in order to not only mimic what viruses and bacterial toxins can do, but also to add new functions, extending or changing the intracellular routes. It is possible to make conjugates or, more preferably, single-chain proteins that recognize cancer cells and deliver cargo inside the cells, even to the desired subcellular compartment. These findings offer new opportunities to deliver drugs/labels only to cancer cells and only to their site of action within the cells. The development of such dual-specificity vectors for targeting cancer cells is an attractive and potentially safer and more efficacious way of delivering drugs. We provide examples of this approach for delivering brain cancer therapeutics, using a specific biomarker on glioblastoma tumor cells. PMID:22671766

  16. Bacterial computing with engineered populations.

    PubMed

    Amos, Martyn; Axmann, Ilka Maria; Blüthgen, Nils; de la Cruz, Fernando; Jaramillo, Alfonso; Rodriguez-Paton, Alfonso; Simmel, Friedrich

    2015-07-28

    We describe strategies for the construction of bacterial computing platforms by describing a number of results from the recently completed bacterial computing with engineered populations project. In general, the implementation of such systems requires a framework containing various components such as intracellular circuits, single cell input/output and cell-cell interfacing, as well as extensive analysis. In this overview paper, we describe our approach to each of these, and suggest possible areas for future research. PMID:26078340

  17. Intracellular Invasion of Orientia tsutsugamushi Activates Inflammasome in ASC-Dependent Manner

    PubMed Central

    Koo, Jung-Eun; Hong, Hye-Jin; Dearth, Andrea; Kobayashi, Koichi S.; Koh, Young-Sang

    2012-01-01

    Orientia tsutsugamushi, a causative agent of scrub typhus, is an obligate intracellular bacterium, which escapes from the endo/phagosome and replicates in the host cytoplasm. O. tsutsugamushi infection induces production of pro-inflammatory mediators including interleukin-1? (IL-1?), which is secreted mainly from macrophages upon cytosolic stimuli by activating cysteine protease caspase-1 within a complex called the inflammasome, and is a key player in initiating and maintaining the inflammatory response. However, the mechanism for IL-1? maturation upon O. tsutsugamushi infection has not been identified. In this study, we show that IL-1 receptor signaling is required for efficient host protection from O. tsutsugamushi infection. Live Orientia, but not heat- or UV-inactivated Orientia, activates the inflammasome through active bacterial uptake and endo/phagosomal maturation. Furthermore, Orientia-stimulated secretion of IL-1? and activation of caspase-1 are ASC- and caspase-1- dependent since IL-1? production was impaired in Asc- and caspase-1-deficient macrophages but not in Nlrp3-, Nlrc4- and Aim2-deficient macrophages. Therefore, live O. tsutsugamushi triggers ASC inflammasome activation leading to IL-1? production, which is a critical innate immune response for effective host defense. PMID:22723924

  18. Macrophage defense mechanisms against intracellular bacteria

    PubMed Central

    Weiss, Günter; Schaible, Ulrich E

    2015-01-01

    Macrophages and neutrophils play a decisive role in host responses to intracellular bacteria including the agent of tuberculosis (TB), Mycobacterium tuberculosis as they represent the forefront of innate immune defense against bacterial invaders. At the same time, these phagocytes are also primary targets of intracellular bacteria to be abused as host cells. Their efficacy to contain and eliminate intracellular M. tuberculosis decides whether a patient initially becomes infected or not. However, when the infection becomes chronic or even latent (as in the case of TB) despite development of specific immune activation, phagocytes have also important effector functions. Macrophages have evolved a myriad of defense strategies to combat infection with intracellular bacteria such as M. tuberculosis. These include induction of toxic anti-microbial effectors such as nitric oxide and reactive oxygen intermediates, the stimulation of microbe intoxication mechanisms via acidification or metal accumulation in the phagolysosome, the restriction of the microbe's access to essential nutrients such as iron, fatty acids, or amino acids, the production of anti-microbial peptides and cytokines, along with induction of autophagy and efferocytosis to eliminate the pathogen. On the other hand, M. tuberculosis, as a prime example of a well-adapted facultative intracellular bacterium, has learned during evolution to counter-balance the host's immune defense strategies to secure survival or multiplication within this otherwise hostile environment. This review provides an overview of innate immune defense of macrophages directed against intracellular bacteria with a focus on M. tuberculosis. Gaining more insights and knowledge into this complex network of host-pathogen interaction will identify novel target sites of intervention to successfully clear infection at a time of rapidly emerging multi-resistance of M. tuberculosis against conventional antibiotics. PMID:25703560

  19. 45 CFR 83.10 - General obligations.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ...2010-10-01 false General obligations. 83...Welfare DEPARTMENT OF HEALTH AND HUMAN SERVICES GENERAL ADMINISTRATION ...AND 845 OF THE PUBLIC HEALTH SERVICE ACT Discrimination...Prohibited § 83.10 General obligations....

  20. 45 CFR 83.10 - General obligations.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ...2011-10-01 false General obligations. 83...Welfare DEPARTMENT OF HEALTH AND HUMAN SERVICES GENERAL ADMINISTRATION ...AND 845 OF THE PUBLIC HEALTH SERVICE ACT Discrimination...Prohibited § 83.10 General obligations....

  1. 38 CFR 17.607 - Obligated service.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ...AFFAIRS MEDICAL Va Health Professional Scholarship Program § 17.607 Obligated service...for which the participant received a scholarship award under these regulations...obligation. A participant who received a scholarship as a full-time student must be...

  2. 7 CFR 400.767 - Requester obligations.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ...2012-01-01 false Requester obligations. 400.767 Section 400.767 Agriculture Regulations of the Department of Agriculture...Interpretations of Statutory and Regulatory Provisions § 400.767 Requester obligations. (a) All requests...

  3. 7 CFR 400.767 - Requester obligations.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ...2010-01-01 false Requester obligations. 400.767 Section 400.767 Agriculture Regulations of the Department of Agriculture...Interpretations of Statutory and Regulatory Provisions § 400.767 Requester obligations. (a) All requests...

  4. 7 CFR 400.767 - Requester obligations.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ...2011-01-01 false Requester obligations. 400.767 Section 400.767 Agriculture Regulations of the Department of Agriculture...Interpretations of Statutory and Regulatory Provisions § 400.767 Requester obligations. (a) All requests...

  5. 7 CFR 400.767 - Requester obligations.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ...2014-01-01 false Requester obligations. 400.767 Section 400.767 Agriculture Regulations of the Department of Agriculture...Interpretations of Statutory and Regulatory Provisions § 400.767 Requester obligations. (a) All requests...

  6. 7 CFR 400.767 - Requester obligations.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ...2013-01-01 false Requester obligations. 400.767 Section 400.767 Agriculture Regulations of the Department of Agriculture...Interpretations of Statutory and Regulatory Provisions § 400.767 Requester obligations. (a) All requests...

  7. 46 CFR Sec. 11 - Guarantee obligations.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ...2011-10-01 2011-10-01 false Guarantee obligations. Sec. 11 Section 11 Shipping MARITIME ADMINISTRATION, DEPARTMENT...AUTHORITY MASTER LUMP SUM REPAIR CONTRACT-NSA-LUMPSUMREP Sec. 11 Guarantee obligations. (a) Under the...

  8. 46 CFR Sec. 11 - Guarantee obligations.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ...2010-10-01 2010-10-01 false Guarantee obligations. Sec. 11 Section 11 Shipping MARITIME ADMINISTRATION, DEPARTMENT...AUTHORITY MASTER LUMP SUM REPAIR CONTRACT-NSA-LUMPSUMREP Sec. 11 Guarantee obligations. (a) Under the...

  9. 7 CFR 993.56 - Reserve obligation.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 8 2010-01-01 2010-01-01 false Reserve obligation. 993.56 Section 993.56 Agriculture...PRODUCED IN CALIFORNIA Order Regulating Handling Reserve Control § 993.56 Reserve obligation. Whenever salable and...

  10. 7 CFR 993.56 - Reserve obligation.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 8 2011-01-01 2011-01-01 false Reserve obligation. 993.56 Section 993.56 Agriculture...PRODUCED IN CALIFORNIA Order Regulating Handling Reserve Control § 993.56 Reserve obligation. Whenever salable and...

  11. Legal obligations of NHS managers.

    PubMed

    Jacks, H J

    1988-10-01

    The loss of crown immunity resulting from section 2 of the National Health Service (Amendment) Act 1986 has understandably caused much concern relating to the application of the Health and Safety at Work Act 1974 to hospitals. The NHS managers must remember that their legal obligations can be both civil and criminal. PMID:10291890

  12. Cell-cycle progress in obligate predatory bacteria is dependent upon sequential sensing of prey recognition and prey quality cues.

    PubMed

    Rotem, Or; Pasternak, Zohar; Shimoni, Eyal; Belausov, Eduard; Porat, Ziv; Pietrokovski, Shmuel; Jurkevitch, Edouard

    2015-11-01

    Predators feed on prey to acquire the nutrients necessary to sustain their survival, growth, and replication. In Bdellovibrio bacteriovorus, an obligate predator of Gram-negative bacteria, cell growth and replication are tied to a shift from a motile, free-living phase of search and attack to a sessile, intracellular phase of growth and replication during which a single prey cell is consumed. Engagement and sustenance of growth are achieved through the sensing of two unidentified prey-derived cues. We developed a novel ex vivo cultivation system for B. bacteriovorus composed of prey ghost cells that are recognized and invaded by the predator. By manipulating their content, we demonstrated that an early cue is located in the prey envelope and a late cue is found within the prey soluble fraction. These spatially and temporally separated cues elicit discrete and combinatory regulatory effects on gene transcription. Together, they delimit a poorly characterized transitory phase between the attack phase and the growth phase, during which the bdelloplast (the invaded prey cell) is constructed. This transitory phase constitutes a checkpoint in which the late cue presumably acts as a determinant of the prey's nutritional value before the predator commits. These regulatory adaptations to a unique bacterial lifestyle have not been reported previously. PMID:26487679

  13. 12 CFR 987.10 - Obligations of United States with respect to consolidated obligations.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 12 Banks and Banking 7 2010-01-01 2010-01-01 false Obligations of United States with respect to consolidated obligations. 987.10 Section 987.10 Banks and Banking FEDERAL HOUSING FINANCE BOARD OFFICE OF FINANCE BOOK-ENTRY PROCEDURE FOR CONSOLIDATED OBLIGATIONS § 987.10 Obligations of United States...

  14. Bacterial gastroenteritis

    MedlinePLUS

    Infectious diarrhea - bacterial gastroenteritis; Acute gastroenteritis; Gastroenteritis - bacterial ... the sickness. All types of food poisoning cause diarrhea . Other symptoms include: Abdominal cramps Abdominal pain Bloody ...

  15. Real-time molecular monitoring of chemical environment in obligate anaerobes during oxygen adaptive response.

    PubMed

    Holman, Hoi-Ying N; Wozei, Eleanor; Lin, Zhang; Comolli, Luis R; Ball, David A; Borglin, Sharon; Fields, Matthew W; Hazen, Terry C; Downing, Kenneth H

    2009-08-01

    Determining the transient chemical properties of the intracellular environment can elucidate the paths through which a biological system adapts to changes in its environment, for example, the mechanisms that enable some obligate anaerobic bacteria to survive a sudden exposure to oxygen. Here we used high-resolution Fourier transform infrared (FTIR) spectromicroscopy to continuously follow cellular chemistry within living obligate anaerobes by monitoring hydrogen bond structures in their cellular water. We observed a sequence of well orchestrated molecular events that correspond to changes in cellular processes in those cells that survive, but only accumulation of radicals in those that do not. We thereby can interpret the adaptive response in terms of transient intracellular chemistry and link it to oxygen stress and survival. This ability to monitor chemical changes at the molecular level can yield important insights into a wide range of adaptive responses. PMID:19541631

  16. Real-Time Molecular Monitoring of Chemical Environment in ObligateAnaerobes during Oxygen Adaptive Response

    SciTech Connect

    Holman, Hoi-Ying N.; Wozei, Eleanor; Lin, Zhang; Comolli, Luis R.; Ball, David. A.; Borglin, Sharon; Fields, Matthew W.; Hazen, Terry C.; Downing, Kenneth H.

    2009-02-25

    Determining the transient chemical properties of the intracellular environment canelucidate the paths through which a biological system adapts to changes in its environment, for example, the mechanisms which enable some obligate anaerobic bacteria to survive a sudden exposure to oxygen. Here we used high-resolution Fourier Transform Infrared (FTIR) spectromicroscopy to continuously follow cellular chemistry within living obligate anaerobes by monitoring hydrogen bonding in their cellular water. We observed a sequence of wellorchestrated molecular events that correspond to changes in cellular processes in those cells that survive, but only accumulation of radicals in those that do not. We thereby can interpret the adaptive response in terms of transient intracellular chemistry and link it to oxygen stress and survival. This ability to monitor chemical changes at the molecular level can yield important insights into a wide range of adaptive responses.

  17. Obligate oil-degrading marine bacteria.

    PubMed

    Yakimov, Michail M; Timmis, Kenneth N; Golyshin, Peter N

    2007-06-01

    Over the past few years, a new and ecophysiologically unusual group of marine hydrocarbon-degrading bacteria - the obligate hydrocarbonoclastic bacteria (OHCB) - has been recognized and shown to play a significant role in the biological removal of petroleum hydrocarbons from polluted marine waters. The introduction of oil or oil constituents into seawater leads to successive blooms of a relatively limited number of indigenous marine bacterial genera--Alcanivorax, Marinobacter, Thallassolituus, Cycloclasticus, Oleispira and a few others (the OHCB)--which are present at low or undetectable levels before the polluting event. The types of OHCB that bloom depend on the latitude/temperature, salinity, redox and other prevailing physical-chemical factors. These blooms result in the rapid degradation of many oil constituents, a process that can be accelerated further by supplementation with limiting nutrients. Genome sequencing and functional genomic analysis of Alcanivorax borkumensis, the paradigm of OHCB, has provided significant insights into the genomic basis of the efficiency and versatility of its hydrocarbon utilization, the metabolic routes underlying its special hydrocarbon diet, and its ecological success. These and other studies have revealed the potential of OHCB for multiple biotechnological applications that include not only oil pollution mitigation, but also biopolymer production and biocatalysis. PMID:17493798

  18. 38 CFR 17.632 - Obligated service.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... Impairment and Orientation and Mobility Professional Scholarship Program § 17.632 Obligated service. (a... service. Geographic relocation may be required. (e) Creditability of advanced clinical training. No...

  19. The Bacterial Cytoskeleton

    PubMed Central

    Shih, Yu-Ling; Rothfield, Lawrence

    2006-01-01

    In recent years it has been shown that bacteria contain a number of cytoskeletal structures. The bacterial cytoplasmic elements include homologs of the three major types of eukaryotic cytoskeletal proteins (actin, tubulin, and intermediate filament proteins) and a fourth group, the MinD-ParA group, that appears to be unique to bacteria. The cytoskeletal structures play important roles in cell division, cell polarity, cell shape regulation, plasmid partition, and other functions. The proteins self-assemble into filamentous structures in vitro and form intracellular ordered structures in vivo. In addition, there are a number of filamentous bacterial elements that may turn out to be cytoskeletal in nature. This review attempts to summarize and integrate the in vivo and in vitro aspects of these systems and to evaluate the probable future directions of this active research field. PMID:16959967

  20. Nanovehicular Intracellular Delivery Systems

    PubMed Central

    PROKOP, ALES; DAVIDSON, JEFFREY M.

    2013-01-01

    This article provides an overview of principles and barriers relevant to intracellular drug and gene transport, accumulation and retention (collectively called as drug delivery) by means of nanovehicles (NV). The aim is to deliver a cargo to a particular intracellular site, if possible, to exert a local action. Some of the principles discussed in this article apply to noncolloidal drugs that are not permeable to the plasma membrane or to the blood–brain barrier. NV are defined as a wide range of nanosized particles leading to colloidal objects which are capable of entering cells and tissues and delivering a cargo intracelullarly. Different localization and targeting means are discussed. Limited discussion on pharmacokinetics and pharmacodynamics is also presented. NVs are contrasted to micro-delivery and current nanotechnologies which are already in commercial use. Newer developments in NV technologies are outlined and future applications are stressed. We also briefly review the existing modeling tools and approaches to quantitatively describe the behavior of targeted NV within the vascular and tumor compartments, an area of particular importance. While we list “elementary” phenomena related to different level of complexity of delivery to cancer, we also stress importance of multi-scale modeling and bottom-up systems biology approach. PMID:18200527

  1. Bacterial Sialidase

    NASA Technical Reports Server (NTRS)

    2004-01-01

    Data shows that elevated sialidase in bacterial vaginosis patients correlates to premature births in women. Bacterial sialidase also plays a significant role in the unusual colonization of Pseudomonas aeruginosa in cystic fibrosis patients. Crystals of Salmonella sialidase have been reproduced and are used for studying the inhibitor-enzyme complexes. These inhibitors may also be used to inhibit a trans-sialidase of Trypanosome cruzi, a very similar enzyme to bacterial sialidase, therefore preventing T. cruzi infection, the causitive agent of Chagas' disease. The Center for Macromolecular Crystallography suggests that inhibitors of bacterial sialidases can be used as prophylactic drugs to prevent bacterial infections in these critical cases.

  2. INTRA-CELLULAR STAPHYLOCOCCUS AUREUS ALONE CAUSES INFECTION IN VIVO#

    PubMed Central

    Hamza, Therwa; Dietz, Matthew; Pham, Danh; Clovis, Nina; Danley, Suzanne; Li, Bingyun

    2013-01-01

    Chronic and recurrent bone infections occur frequently but have not been explained. Staphylococcus aureus (S. aureus) is often found among chronic and recurrent infections and may be responsible for such infections. One possible reason is that S. aureus can internalize and survive within host cells and by doing so, S. aureus can evade both host defense mechanisms and most conventional antibiotic treatments. In this study, we hypothesized that intra-cellular S. aureus could induce infections in vivo. Osteoblasts were infected with S. aureus and, after eliminating extra-cellular S. aureus, inoculated into an open fracture rat model. Bacterial cultures and radiographic observations at post-operative day 21 confirmed local bone infections in animals inoculated with intra-cellular S. aureus within osteoblasts alone. We present direct in vivo evidence that intra-cellular S. aureus could be sufficient to induce bone infection in animals; we found that intra-cellular S. aureus inoculation of as low as 102 colony forming units could induce severe bone infections. Our data may suggest that intra-cellular S. aureus can “hide” in host cells during symptom-free periods and, under certain conditions, they may escape and lead to infection recurrence. Intra-cellular S. aureus therefore could play an important role in the pathogenesis of S. aureus infections, especially those chronic and recurrent infections in which disease episodes may be separated by weeks, months, or even years. PMID:23832687

  3. Divine voluntarism: moral obligation supervenes on God's antecedent will 

    E-print Network

    Nam, Mi Young

    2004-11-15

    's will for human moral obligation and God's will for human moral good. After all, God's will for human moral obligation is God's willing that His own will for human moral good constitute moral obligation for humans....

  4. 46 CFR 298.30 - Nature and content of Obligations.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ...46 Shipping 8 2010-10-01 2010-10-01 false Nature and content of Obligations. 298.30 Section 298.30...ASSISTANCE OBLIGATION GUARANTEES Documentation § 298.30 Nature and content of Obligations. (a) Single page....

  5. 46 CFR 298.30 - Nature and content of Obligations.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ...46 Shipping 8 2011-10-01 2011-10-01 false Nature and content of Obligations. 298.30 Section 298.30...ASSISTANCE OBLIGATION GUARANTEES Documentation § 298.30 Nature and content of Obligations. (a) Single page....

  6. 31 CFR 225.5 - Pledge of definitive Government obligations.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... false Pledge of definitive Government obligations. 225.5 Section...ACCEPTANCE OF BONDS SECURED BY GOVERNMENT OBLIGATIONS IN LIEU OF BONDS...225.5 Pledge of definitive Government obligations. (a) Type...

  7. 31 CFR 225.5 - Pledge of definitive Government obligations.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... false Pledge of definitive Government obligations. 225.5 Section...ACCEPTANCE OF BONDS SECURED BY GOVERNMENT OBLIGATIONS IN LIEU OF BONDS...225.5 Pledge of definitive Government obligations. (a) Type...

  8. Bacterial Proteasomes

    PubMed Central

    Jastrab, Jordan B.; Darwin, K. Heran

    2015-01-01

    Interest in bacterial proteasomes was sparked by the discovery that proteasomal degradation is required for the pathogenesis of Mycobacterium tuberculosis, one of the world's deadliest pathogens. Although bacterial proteasomes are structurally similar to their eukaryotic and archaeal homologs, there are key differences in their mechanisms of assembly, activation, and substrate targeting for degradation. In this article, we compare and contrast bacterial proteasomes with their archaeal and eukaryotic counterparts, and we discuss recent advances in our understanding of how bacterial proteasomes function to influence microbial physiology. PMID:26488274

  9. Exit Mechanisms of the Intracellular Bacterium Ehrlichia

    PubMed Central

    Thomas, Sunil; Popov, Vsevolod L.; Walker, David H.

    2010-01-01

    Background The obligately intracellular bacterium Ehrlichia chaffeensis that resides in mononuclear phagocytes is the causative agent of human monocytotropic ehrlichiosis. Ehrlichia muris and Ixodes ovatus Ehrlichia (IOE) are agents of mouse models of ehrlichiosis. The mechanism by which Ehrlichia are transported from an infected host cell to a non-infected cell has not been demonstrated. Methodology/Principal Findings Using fluorescence microscopy and transmission and scanning electron microscopy, we demonstrated that Ehrlichia was transported through the filopodia of macrophages during early stages of infection. If host cells were not present in the vicinity of an Ehrlichia-infected cell, the leading edge of the filopodium formed a fan-shaped structure filled with the pathogen. Formation of filopodia in the host macrophages was inhibited by cytochalasin D and ehrlichial transport were prevented due to the absence of filopodia formation. At late stages of infection the host cell membrane was ruptured, and the bacteria were released. Conclusions/Significance Ehrlichia are transported through the host cell filopodium during initial stages of infection, but are released by host cell membrane rupture during later stages of infection. PMID:21187937

  10. Evaluation of the bacterial microbiome of two flea species using different DNA-isolation techniques provides insights into flea host ecology.

    PubMed

    Lawrence, Andrea L; Hii, Sze-Fui; Chong, Rowena; Webb, Cameron E; Traub, Rebecca; Brown, Graeme; Šlapeta, Jan

    2015-12-01

    Fleas (Siphonaptera) are ubiquitous blood-sucking pests of animals worldwide and are vectors of zoonotic bacteria such as Rickettsia and Bartonella. We performed Ion Torrent PGM amplicon sequencing for the bacterial 16S rRNA gene to compare the microbiome of the ubiquitous cat flea (Ctenocephalides f. felis) and the host-specific echidna stickfast flea (Echidnophaga a. ambulans) and evaluated potential bias produced during common genomic DNA-isolation methods. We demonstrated significant differences in the bacterial community diversity between the two flea species but not between protocols combining surface sterilisation with whole flea homogenisation or exoskeleton retention. Both flea species were dominated by obligate intracellular endosymbiont Wolbachia, and the echidna stickfast fleas possessed the endosymbiont Cardinium. Cat fleas that were not surface sterilised showed presence of Candidatus 'Rickettsia senegalensis' DNA, the first report of its presence in Australia. In the case of Rickettsia, we show that sequencing depth of 50 000 was required for comparable sensitivity with Rickettsia qPCR. Low-abundance bacterial genera are suggested to reflect host ecology. The deep-sequencing approach demonstrates feasibility of pathogen detection with simultaneous quantitative analysis and evaluation of the inter-relationship of microbes within vectors. PMID:26542076

  11. Bacterial Exotoxins and the Inflammasome

    PubMed Central

    Greaney, Allison J.; Leppla, Stephen H.; Moayeri, Mahtab

    2015-01-01

    The inflammasomes are intracellular protein complexes that play an important role in innate immune sensing. Activation of inflammasomes leads to activation of caspase-1 and maturation and secretion of the pro-inflammatory cytokines interleukin (IL)-1? and IL-18. In certain myeloid cells, this activation can also lead to an inflammatory cell death (pyroptosis). Inflammasome sensor proteins have evolved to detect a range of microbial ligands and bacterial exotoxins either through direct interaction or by detection of host cell changes elicited by these effectors. Bacterial exotoxins activate the inflammasomes through diverse processes, including direct sensor cleavage, modulation of ion fluxes through plasma membrane pore formation, and perturbation of various host cell functions. In this review, we summarize the findings on some of the bacterial exotoxins that activate the inflammasomes. PMID:26617605

  12. High-Throughput Intracellular Antimicrobial Susceptibility Testing of Legionella pneumophila.

    PubMed

    Chiaraviglio, Lucius; Kirby, James E

    2015-12-01

    Legionella pneumophila is a Gram-negative opportunistic human pathogen that causes a severe pneumonia known as Legionnaires' disease. Notably, in the human host, the organism is believed to replicate solely within an intracellular compartment, predominantly within pulmonary macrophages. Consequently, successful therapy is predicated on antimicrobials penetrating into this intracellular growth niche. However, standard antimicrobial susceptibility testing methods test solely for extracellular growth inhibition. Here, we make use of a high-throughput assay to characterize intracellular growth inhibition activity of known antimicrobials. For select antimicrobials, high-resolution dose-response analysis was then performed to characterize and compare activity levels in both macrophage infection and axenic growth assays. Results support the superiority of several classes of nonpolar antimicrobials in abrogating intracellular growth. Importantly, our assay results show excellent correlations with prior clinical observations of antimicrobial efficacy. Furthermore, we also show the applicability of high-throughput automation to two- and three-dimensional synergy testing. High-resolution isocontour isobolograms provide in vitro support for specific combination antimicrobial therapy. Taken together, findings suggest that high-throughput screening technology may be successfully applied to identify and characterize antimicrobials that target bacterial pathogens that make use of an intracellular growth niche. PMID:26392509

  13. 46 CFR Sec. 11 - Guarantee obligations.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... ACCOMPLISHMENT OF VESSEL REPAIRS UNDER NATIONAL SHIPPING AUTHORITY MASTER LUMP SUM REPAIR CONTRACT-NSA-LUMPSUMREP Sec. 11 Guarantee obligations. (a) Under the provisions of Article 10 of the NSA-LUMPSUMREP...

  14. 46 CFR Sec. 11 - Guarantee obligations.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... ACCOMPLISHMENT OF VESSEL REPAIRS UNDER NATIONAL SHIPPING AUTHORITY MASTER LUMP SUM REPAIR CONTRACT-NSA-LUMPSUMREP Sec. 11 Guarantee obligations. (a) Under the provisions of Article 10 of the NSA-LUMPSUMREP...

  15. 46 CFR Sec. 11 - Guarantee obligations.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... ACCOMPLISHMENT OF VESSEL REPAIRS UNDER NATIONAL SHIPPING AUTHORITY MASTER LUMP SUM REPAIR CONTRACT-NSA-LUMPSUMREP Sec. 11 Guarantee obligations. (a) Under the provisions of Article 10 of the NSA-LUMPSUMREP...

  16. 46 CFR Sec. 11 - Guarantee obligations.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... ACCOMPLISHMENT OF VESSEL REPAIRS UNDER NATIONAL SHIPPING AUTHORITY MASTER LUMP SUM REPAIR CONTRACT-NSA-LUMPSUMREP Sec. 11 Guarantee obligations. (a) Under the provisions of Article 10 of the NSA-LUMPSUMREP...

  17. 5 CFR 724.103 - Agency obligations.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ...Agency obligations. A Federal agency (or its successor agency) must reimburse the Judgment Fund for payments covered by the No FEAR Act. Such reimbursement must be made within a reasonable time as described in §...

  18. 42 CFR 408.4 - Payment obligations.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... kidney donors. (1) No premiums are required for SMI benefits related to the donation of a kidney if the donor is not an enrollee. (2) A kidney donor who is an enrollee is not relieved of the obligation...

  19. 42 CFR 408.4 - Payment obligations.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... kidney donors. (1) No premiums are required for SMI benefits related to the donation of a kidney if the donor is not an enrollee. (2) A kidney donor who is an enrollee is not relieved of the obligation...

  20. 42 CFR 408.4 - Payment obligations.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... kidney donors. (1) No premiums are required for SMI benefits related to the donation of a kidney if the donor is not an enrollee. (2) A kidney donor who is an enrollee is not relieved of the obligation...

  1. 42 CFR 408.4 - Payment obligations.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... kidney donors. (1) No premiums are required for SMI benefits related to the donation of a kidney if the donor is not an enrollee. (2) A kidney donor who is an enrollee is not relieved of the obligation...

  2. 42 CFR 408.4 - Payment obligations.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... kidney donors. (1) No premiums are required for SMI benefits related to the donation of a kidney if the donor is not an enrollee. (2) A kidney donor who is an enrollee is not relieved of the obligation...

  3. 19 CFR 10.765 - Importer obligations.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...HOMELAND SECURITY; DEPARTMENT OF THE TREASURY ARTICLES CONDITIONALLY FREE, SUBJECT TO A REDUCED RATE, ETC. United States-Morocco Free Trade Agreement Import Requirements § 10.765 Importer obligations. (a) General . An importer who...

  4. 45 CFR 2400.65 - Teaching obligation.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ...Teaching obligation. 2400.65 Section 2400.65 Public Welfare Regulations Relating to Public Welfare (Continued) JAMES MADISON MEMORIAL FELLOWSHIP FOUNDATION FELLOWSHIP PROGRAM REQUIREMENTS Special Conditions § 2400.65 Teaching...

  5. 45 CFR 2400.65 - Teaching obligation.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ...Teaching obligation. 2400.65 Section 2400.65 Public Welfare Regulations Relating to Public Welfare (Continued) JAMES MADISON MEMORIAL FELLOWSHIP FOUNDATION FELLOWSHIP PROGRAM REQUIREMENTS Special Conditions § 2400.65 Teaching...

  6. 45 CFR 2400.65 - Teaching obligation.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ...Teaching obligation. 2400.65 Section 2400.65 Public Welfare Regulations Relating to Public Welfare (Continued) JAMES MADISON MEMORIAL FELLOWSHIP FOUNDATION FELLOWSHIP PROGRAM REQUIREMENTS Special Conditions § 2400.65 Teaching...

  7. 45 CFR 2400.65 - Teaching obligation.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ...Teaching obligation. 2400.65 Section 2400.65 Public Welfare Regulations Relating to Public Welfare (Continued) JAMES MADISON MEMORIAL FELLOWSHIP FOUNDATION FELLOWSHIP PROGRAM REQUIREMENTS Special Conditions § 2400.65 Teaching...

  8. 45 CFR 2400.65 - Teaching obligation.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ...Teaching obligation. 2400.65 Section 2400.65 Public Welfare Regulations Relating to Public Welfare (Continued) JAMES MADISON MEMORIAL FELLOWSHIP FOUNDATION FELLOWSHIP PROGRAM REQUIREMENTS Special Conditions § 2400.65 Teaching...

  9. Naval Engineering A National Naval Obligation

    E-print Network

    Chryssostomidis, Chryssostomos

    2000-05-16

    As part of its national obligations, ONR must ensure US world leadership in those unique technology areas that insure naval superiority. ONR accomplishes this mission through research, recruitment and education, maintaining ...

  10. 5 CFR 724.404 - Agency obligations.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ...CONTINUED) IMPLEMENTATION OF TITLE II OF THE NOTIFICATION AND FEDERAL EMPLOYEE ANTIDISCRIMINATION AND RETALIATION ACT OF 2002 Best Practices § 724.404 Agency obligations. (a) Within 30 working days of issuance of the advisory...

  11. 46 CFR Sec. 11 - Guarantee obligations.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... ACCOMPLISHMENT OF VESSEL REPAIRS UNDER NATIONAL SHIPPING AUTHORITY MASTER LUMP SUM REPAIR CONTRACT-NSA-LUMPSUMREP Sec. 11 Guarantee obligations. (a) Under the provisions of Article 10 of the NSA-LUMPSUMREP...

  12. 7 CFR 989.37 - Obligation.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ...MARKETING SERVICE (Marketing Agreements and Orders; Fruits, Vegetables, Nuts), DEPARTMENT OF AGRICULTURE RAISINS PRODUCED FROM GRAPES GROWN IN CALIFORNIA Order Regulating Handling Raisin Administrative Committee § 989.37 Obligation....

  13. 46 CFR 298.30 - Nature and content of Obligations.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 8 2011-10-01 2011-10-01 false Nature and content of Obligations. 298.30 Section 298.30 Shipping MARITIME ADMINISTRATION, DEPARTMENT OF TRANSPORTATION VESSEL FINANCING ASSISTANCE OBLIGATION GUARANTEES Documentation § 298.30 Nature and content of Obligations. (a) Single page. An Obligation, in...

  14. 46 CFR 298.30 - Nature and content of Obligations.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 46 Shipping 8 2013-10-01 2013-10-01 false Nature and content of Obligations. 298.30 Section 298.30 Shipping MARITIME ADMINISTRATION, DEPARTMENT OF TRANSPORTATION VESSEL FINANCING ASSISTANCE OBLIGATION GUARANTEES Documentation § 298.30 Nature and content of Obligations. (a) Single page. An Obligation, in...

  15. 46 CFR 298.30 - Nature and content of Obligations.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 8 2014-10-01 2014-10-01 false Nature and content of Obligations. 298.30 Section 298.30 Shipping MARITIME ADMINISTRATION, DEPARTMENT OF TRANSPORTATION VESSEL FINANCING ASSISTANCE OBLIGATION GUARANTEES Documentation § 298.30 Nature and content of Obligations. (a) Single page. An Obligation, in...

  16. 46 CFR 298.30 - Nature and content of Obligations.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 8 2010-10-01 2010-10-01 false Nature and content of Obligations. 298.30 Section 298.30 Shipping MARITIME ADMINISTRATION, DEPARTMENT OF TRANSPORTATION VESSEL FINANCING ASSISTANCE OBLIGATION GUARANTEES Documentation § 298.30 Nature and content of Obligations. (a) Single page. An Obligation, in...

  17. 46 CFR 298.30 - Nature and content of Obligations.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 8 2012-10-01 2012-10-01 false Nature and content of Obligations. 298.30 Section 298.30 Shipping MARITIME ADMINISTRATION, DEPARTMENT OF TRANSPORTATION VESSEL FINANCING ASSISTANCE OBLIGATION GUARANTEES Documentation § 298.30 Nature and content of Obligations. (a) Single page. An Obligation, in...

  18. Benefit Finding and Perceived Obligations of Victims

    E-print Network

    Warner, Ruth

    2007-12-11

    a mention of physical ilnes, mental ilnes, abuse, drug addiction, or obesity. They found that participants felt more anger and les pity for the poor copers as compared to the good copers. They also offered les social support to the poor coper. How... and target of leson were used to predict victim obligations, benefit finding significantly predicted victim obligations, ? = .51, p test was significant, Z = 2...

  19. Bacterial Gut Symbionts Contribute to Seed Digestion in an Omnivorous Beetle

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Obligate bacterial symbionts alter the diets of host animals in numerous ways, but the ecological roles of facultative bacterial residents that consistently colonize insect guts remain unclear. Carabid beetles are a common group of beneficial insects appreciated for their ability to consume insect p...

  20. Lateral phage transfer in obligate intracellular bacteria (Wolbachia): Verification from natural populations

    E-print Network

    Bordenstein, Seth

    , of Gryllus pennsylvanicus field crickets and of Neochlamisus bebbianae leaf beetles, to demonstrate WO and horizontally have dynamic genomes exhibiting phage transfer, recombination, and toxin-gene acquisitions (Oliver

  1. INTRACELLULAR SIGNALING AND DEVELOPMENTAL NEUROTOXICITY.

    EPA Science Inventory

    A book chapter in ?Molecular Toxicology: Transcriptional Targets? reviewed the role of intracellular signaling in the developmental neurotoxicity of environmental chemicals. This chapter covered a number of aspects including the development of the nervous system, role of intrace...

  2. Intracellular Adaptation of Brucella abortus

    PubMed Central

    Lamontagne, Julie; Forest, Anik; Marazzo, Elena; Denis, François; Butler, Heather; Michaud, Jean-François; Boucher, Lyne; Pedro, Ida; Villeneuve, Annie; Sitnikov, Dmitri; Trudel, Karine; Nassif, Najib; Boudjelti, Djamila; Tomaki, Fadi; Chaves-Olarte, Esteban; Guzmán-Verri, Caterina; Brunet, Sylvain; Côté-Martin, Alexandra; Hunter, Joanna; Moreno, Edgardo; Paramithiotis, Eustache

    2009-01-01

    Macrophages were infected with virulent B. abortus strain 2308 or attenuated strain 19. Intracellular bacteria were recovered at different times after infection and their proteomes compared. The virulent strain initially reduced most biosynthesis and altered its respiration, adaptations reversed later in infection. The attenuated strain was unable to match the magnitude of the virulent strain’s adjustments. The results provide insight into mechanisms utilized by Brucella to establish intracellular infections. PMID:19216536

  3. Modulation of p53 during bacterial infections.

    PubMed

    Siegl, Christine; Rudel, Thomas

    2015-12-01

    In recent years, numerous bacterial pathogens have been shown to inactivate the major tumour suppressor p53 during infection. This inactivation impedes the protective response of the host cell to the genotoxicity that often results from bacterial infection. Moreover, a new aspect of the antibacterial activity of p53 that has recently come to light - downregulation of host cell metabolism to interfere with intracellular bacterial replication - has further highlighted the crucial role of p53 in host-pathogen interactions, as host cell metabolism is relevant for all intracellular bacteria, as well as other pathogens that replicate inside host cells and use host metabolites. In this Progress article, we summarize recent work that has advanced our knowledge of the interaction between pathogenic bacteria and p53, and we discuss the known and expected outcomes of this interaction for pathogenesis. PMID:26548915

  4. Understanding how commensal obligate anaerobic bacteria regulate immune functions in the large intestine.

    PubMed

    Maier, Eva; Anderson, Rachel C; Roy, Nicole C

    2015-01-01

    The human gastrointestinal tract is colonised by trillions of commensal bacteria, most of which are obligate anaerobes residing in the large intestine. Appropriate bacterial colonisation is generally known to be critical for human health. In particular, the development and function of the immune system depends on microbial colonisation, and a regulated cross-talk between commensal bacteria, intestinal epithelial cells and immune cells is required to maintain mucosal immune homeostasis. This homeostasis is disturbed in various inflammatory disorders, such as inflammatory bowel diseases. Several in vitro and in vivo studies indicate a role for Faecalibacterium prausnitzii, Bacteroides thetaiotaomicron, Bacteroides fragilis, Akkermansia muciniphila and segmented filamentous bacteria in maintaining intestinal immune homeostasis. These obligate anaerobes are abundant in the healthy intestine but reduced in several inflammatory diseases, suggesting an association with protective effects on human health. However, knowledge of the mechanisms underlying the effects of obligate anaerobic intestinal bacteria remains limited, in part due to the difficulty of co-culturing obligate anaerobes together with oxygen-requiring human epithelial cells. By using novel dual-environment co-culture models, it will be possible to investigate the effects of the unstudied majority of intestinal microorganisms on the human epithelia. This knowledge will provide opportunities for improving human health and reducing the risk of inflammatory diseases. PMID:25545102

  5. Understanding How Commensal Obligate Anaerobic Bacteria Regulate Immune Functions in the Large Intestine

    PubMed Central

    Maier, Eva; Anderson, Rachel C.; Roy, Nicole C.

    2014-01-01

    The human gastrointestinal tract is colonised by trillions of commensal bacteria, most of which are obligate anaerobes residing in the large intestine. Appropriate bacterial colonisation is generally known to be critical for human health. In particular, the development and function of the immune system depends on microbial colonisation, and a regulated cross-talk between commensal bacteria, intestinal epithelial cells and immune cells is required to maintain mucosal immune homeostasis. This homeostasis is disturbed in various inflammatory disorders, such as inflammatory bowel diseases. Several in vitro and in vivo studies indicate a role for Faecalibacterium prausnitzii, Bacteroides thetaiotaomicron, Bacteroides fragilis, Akkermansia muciniphila and segmented filamentous bacteria in maintaining intestinal immune homeostasis. These obligate anaerobes are abundant in the healthy intestine but reduced in several inflammatory diseases, suggesting an association with protective effects on human health. However, knowledge of the mechanisms underlying the effects of obligate anaerobic intestinal bacteria remains limited, in part due to the difficulty of co-culturing obligate anaerobes together with oxygen-requiring human epithelial cells. By using novel dual-environment co-culture models, it will be possible to investigate the effects of the unstudied majority of intestinal microorganisms on the human epithelia. This knowledge will provide opportunities for improving human health and reducing the risk of inflammatory diseases. PMID:25545102

  6. Whistleblowing and the bioethicist's public obligations.

    PubMed

    MacDougall, D Robert

    2014-10-01

    Bioethicists are sometimes thought to have heightened obligations by virtue of the fact that their professional role addresses ethics or morals. For this reason it has been argued that bioethicists ought to "whistleblow"--that is, publicly expose the wrongful or potentially harmful activities of their employer--more often than do other kinds of employees. This article argues that bioethicists do indeed have a heightened obligation to whistleblow, but not because bioethicists have heightened moral obligations in general. Rather, the special duties of bioethicists to act as whistleblowers are best understood by examining the nature of the ethical dilemma typically encountered by private employees and showing why bioethicists do not encounter this dilemma in the same way. Whistleblowing is usually understood as a moral dilemma involving conflicting duties to two parties: the public and a private employer. However, this article argues that this way of understanding whistleblowing has the implication that professions whose members identify their employer as the public-such as government employees or public servants--cannot consider whistleblowing a moral dilemma, because obligations are ultimately owed to only one party: the public. The article contends that bioethicists--even when privately employed--are similar to government employees in the sense that they do not have obligations to defer to the judgments of those with private interests. Consequently, bioethicists may be considered to have a special duty to whistleblow, although for different reasons than those usually cited. PMID:25045940

  7. Comparative Genomics of Wolbachia and the Bacterial Species Concept

    PubMed Central

    Näslund, Kristina; Bourtzis, Kostas; Andersson, Siv G. E.

    2013-01-01

    The importance of host-specialization to speciation processes in obligate host-associated bacteria is well known, as is also the ability of recombination to generate cohesion in bacterial populations. However, whether divergent strains of highly recombining intracellular bacteria, such as Wolbachia, can maintain their genetic distinctness when infecting the same host is not known. We first developed a protocol for the genome sequencing of uncultivable endosymbionts. Using this method, we have sequenced the complete genomes of the Wolbachia strains wHa and wNo, which occur as natural double infections in Drosophila simulans populations on the Seychelles and in New Caledonia. Taxonomically, wHa belong to supergroup A and wNo to supergroup B. A comparative genomics study including additional strains supported the supergroup classification scheme and revealed 24 and 33 group-specific genes, putatively involved in host-adaptation processes. Recombination frequencies were high for strains of the same supergroup despite different host-preference patterns, leading to genomic cohesion. The inferred recombination fragments for strains of different supergroups were of short sizes, and the genomes of the co-infecting Wolbachia strains wHa and wNo were not more similar to each other and did not share more genes than other A- and B-group strains that infect different hosts. We conclude that Wolbachia strains of supergroup A and B represent genetically distinct clades, and that strains of different supergroups can co-exist in the same arthropod host without converging into the same species. This suggests that the supergroups are irreversibly separated and that barriers other than host-specialization are able to maintain distinct clades in recombining endosymbiont populations. Acquiring a good knowledge of the barriers to genetic exchange in Wolbachia will advance our understanding of how endosymbiont communities are constructed from vertically and horizontally transmitted genes. PMID:23593012

  8. Nutrient salvaging and metabolism by the intracellular pathogen Legionella pneumophila

    PubMed Central

    Fonseca, Maris V.; Swanson, Michele S.

    2014-01-01

    The Gram-negative bacterium Legionella pneumophila is ubiquitous in freshwater environments as a free-swimming organism, resident of biofilms, or parasite of protozoa. If the bacterium is aerosolized and inhaled by a susceptible human host, it can infect alveolar macrophages and cause a severe pneumonia known as Legionnaires' disease. A sophisticated cell differentiation program equips L. pneumophila to persist in both extracellular and intracellular niches. During its life cycle, L. pneumophila alternates between at least two distinct forms: a transmissive form equipped to infect host cells and evade lysosomal degradation, and a replicative form that multiplies within a phagosomal compartment that it has retooled to its advantage. The efficient changeover between transmissive and replicative states is fundamental to L. pneumophila's fitness as an intracellular pathogen. The transmission and replication programs of L. pneumophila are governed by a number of metabolic cues that signal whether conditions are favorable for replication or instead trigger escape from a spent host. Several lines of experimental evidence gathered over the past decade establish strong links between metabolism, cellular differentiation, and virulence of L. pneumophila. Herein, we focus on current knowledge of the metabolic components employed by intracellular L. pneumophila for cell differentiation, nutrient salvaging and utilization of host factors. Specifically, we highlight the metabolic cues that are coupled to bacterial differentiation, nutrient acquisition systems, and the strategies utilized by L. pneumophila to exploit host metabolites for intracellular replication. PMID:24575391

  9. Relative entropy differences in bacterial chromosomes, plasmids, phages and genomic islands

    PubMed Central

    2012-01-01

    Background We sought to assess whether the concept of relative entropy (information capacity), could aid our understanding of the process of horizontal gene transfer in microbes. We analyzed the differences in information capacity between prokaryotic chromosomes, genomic islands (GI), phages, and plasmids. Relative entropy was estimated using the Kullback-Leibler measure. Results Relative entropy was highest in bacterial chromosomes and had the sequence chromosomes > GI > phage > plasmid. There was an association between relative entropy and AT content in chromosomes, phages, plasmids and GIs with the strongest association being in phages. Relative entropy was also found to be lower in the obligate intracellular Mycobacterium leprae than in the related M. tuberculosis when measured on a shared set of highly conserved genes. Conclusions We argue that relative entropy differences reflect how plasmids, phages and GIs interact with microbial host chromosomes and that all these biological entities are, or have been, subjected to different selective pressures. The rate at which amelioration of horizontally acquired DNA occurs within the chromosome is likely to account for the small differences between chromosomes and stably incorporated GIs compared to the transient or independent replicons such as phages and plasmids. PMID:22325062

  10. Magnetic microbes: Bacterial magnetite biomineralization

    SciTech Connect

    Prozorov, Tanya

    2015-09-14

    Magnetotactic bacteria are a diverse group of prokaryotes with the ability to orient and migrate along the magnetic field lines in search for a preferred oxygen concentration in chemically stratified water columns and sediments. These microorganisms produce magnetosomes, the intracellular nanometer-sized magnetic crystals surrounded by a phospholipid bilayer membrane, typically organized in chains. Magnetosomes have nearly perfect crystal structures with narrow size distribution and species-specific morphologies, leading to well-defined magnetic properties. As a result, the magnetite biomineralization in these organisms is of fundamental interest to diverse disciplines, from biotechnology to astrobiology. As a result, this article highlights recent advances in the understanding of the bacterial magnetite biomineralization.

  11. Magnetic microbes: Bacterial magnetite biomineralization.

    PubMed

    Prozorov, Tanya

    2015-10-01

    Magnetotactic bacteria are a diverse group of prokaryotes with the ability to orient and migrate along the magnetic field lines in search for a preferred oxygen concentration in chemically stratified water columns and sediments. These microorganisms produce magnetosomes, the intracellular nanometer-sized magnetic crystals surrounded by a phospholipid bilayer membrane, typically organized in chains. Magnetosomes have nearly perfect crystal structures with narrow size distribution and species-specific morphologies, leading to well-defined magnetic properties. As a result, the magnetite biomineralization in these organisms is of fundamental interest to diverse disciplines, from biotechnology to astrobiology. This article highlights recent advances in the understanding of the bacterial magnetite biomineralization. PMID:26382301

  12. Importance of Branched-Chain Amino Acid Utilization in Francisella Intracellular Adaptation

    PubMed Central

    Gesbert, Gael; Ramond, Elodie; Tros, Fabiola; Dairou, Julien; Frapy, Eric; Barel, Monique

    2014-01-01

    Intracellular bacterial pathogens have adapted their metabolism to optimally utilize the nutrients available in infected host cells. We recently reported the identification of an asparagine transporter required specifically for cytosolic multiplication of Francisella. In the present work, we characterized a new member of the major super family (MSF) of transporters, involved in isoleucine uptake. We show that this transporter (here designated IleP) plays a critical role in intracellular metabolic adaptation of Francisella. Inactivation of IleP severely impaired intracellular F. tularensis subsp. novicida multiplication in all cell types tested and reduced bacterial virulence in the mouse model. To further establish the importance of the ileP gene in F. tularensis pathogenesis, we constructed a chromosomal deletion mutant of ileP (?FTL_1803) in the F. tularensis subsp. holarctica live vaccine strain (LVS). Inactivation of IleP in the F. tularensis LVS provoked comparable intracellular growth defects, confirming the critical role of this transporter in isoleucine uptake. The data presented establish, for the first time, the importance of isoleucine utilization for efficient phagosomal escape and cytosolic multiplication of Francisella and suggest that virulent F. tularensis subspecies have lost their branched-chain amino acid biosynthetic pathways and rely exclusively on dedicated uptake systems. This loss of function is likely to reflect an evolution toward a predominantly intracellular life style of the pathogen. Amino acid transporters should be thus considered major players in the adaptation of intracellular pathogens. PMID:25332124

  13. Importance of branched-chain amino acid utilization in Francisella intracellular adaptation.

    PubMed

    Gesbert, Gael; Ramond, Elodie; Tros, Fabiola; Dairou, Julien; Frapy, Eric; Barel, Monique; Charbit, Alain

    2015-01-01

    Intracellular bacterial pathogens have adapted their metabolism to optimally utilize the nutrients available in infected host cells. We recently reported the identification of an asparagine transporter required specifically for cytosolic multiplication of Francisella. In the present work, we characterized a new member of the major super family (MSF) of transporters, involved in isoleucine uptake. We show that this transporter (here designated IleP) plays a critical role in intracellular metabolic adaptation of Francisella. Inactivation of IleP severely impaired intracellular F. tularensis subsp. novicida multiplication in all cell types tested and reduced bacterial virulence in the mouse model. To further establish the importance of the ileP gene in F. tularensis pathogenesis, we constructed a chromosomal deletion mutant of ileP (?FTL_1803) in the F. tularensis subsp. holarctica live vaccine strain (LVS). Inactivation of IleP in the F. tularensis LVS provoked comparable intracellular growth defects, confirming the critical role of this transporter in isoleucine uptake. The data presented establish, for the first time, the importance of isoleucine utilization for efficient phagosomal escape and cytosolic multiplication of Francisella and suggest that virulent F. tularensis subspecies have lost their branched-chain amino acid biosynthetic pathways and rely exclusively on dedicated uptake systems. This loss of function is likely to reflect an evolution toward a predominantly intracellular life style of the pathogen. Amino acid transporters should be thus considered major players in the adaptation of intracellular pathogens. PMID:25332124

  14. 12 CFR 966.2 - Issuance of consolidated obligations.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ...Section 966.2 Banks and Banking FEDERAL HOUSING FINANCE BOARD FEDERAL HOME LOAN BANK LIABILITIES CONSOLIDATED... (a) Consolidated obligations issued by the Finance Board. The Finance Board may issue consolidated obligations...

  15. 12 CFR 966.2 - Issuance of consolidated obligations.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ...Section 966.2 Banks and Banking FEDERAL HOUSING FINANCE BOARD FEDERAL HOME LOAN BANK LIABILITIES CONSOLIDATED... (a) Consolidated obligations issued by the Finance Board. The Finance Board may issue consolidated obligations...

  16. 22 CFR 221.15 - Fiscal Agent obligations.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ...false Fiscal Agent obligations. 221.15 Section 221.15 Foreign Relations AGENCY FOR INTERNATIONAL DEVELOPMENT ISRAEL LOAN GUARANTEE STANDARD TERMS AND CONDITIONS The Guarantee § 221.15 Fiscal Agent obligations. Failure of the...

  17. 22 CFR 221.15 - Fiscal Agent obligations.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ...false Fiscal Agent obligations. 221.15 Section 221.15 Foreign Relations AGENCY FOR INTERNATIONAL DEVELOPMENT ISRAEL LOAN GUARANTEE STANDARD TERMS AND CONDITIONS The Guarantee § 221.15 Fiscal Agent obligations. Failure of the...

  18. 22 CFR 221.15 - Fiscal Agent obligations.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...false Fiscal Agent obligations. 221.15 Section 221.15 Foreign Relations AGENCY FOR INTERNATIONAL DEVELOPMENT ISRAEL LOAN GUARANTEE STANDARD TERMS AND CONDITIONS The Guarantee § 221.15 Fiscal Agent obligations. Failure of the...

  19. 22 CFR 221.15 - Fiscal Agent obligations.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...false Fiscal Agent obligations. 221.15 Section 221.15 Foreign Relations AGENCY FOR INTERNATIONAL DEVELOPMENT ISRAEL LOAN GUARANTEE STANDARD TERMS AND CONDITIONS The Guarantee § 221.15 Fiscal Agent obligations. Failure of the...

  20. 22 CFR 221.15 - Fiscal Agent obligations.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ...false Fiscal Agent obligations. 221.15 Section 221.15 Foreign Relations AGENCY FOR INTERNATIONAL DEVELOPMENT ISRAEL LOAN GUARANTEE STANDARD TERMS AND CONDITIONS The Guarantee § 221.15 Fiscal Agent obligations. Failure of the...

  1. 12 CFR 560.42 - State and local government obligations.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ...Federal Savings Associations § 560.42 State and local government obligations. (a) What limitations apply? Pursuant to HOLA section 5(c)(1)(H), a Federal savings association (“you”) may invest in obligations issued by any state,...

  2. 5 CFR 2635.101 - Basic obligation of public service.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 2011-01-01 false Basic obligation of public service. 2635.101 Section 2635.101 Administrative...Provisions § 2635.101 Basic obligation of public service. (a) Public service is a public trust. Each...

  3. 5 CFR 2635.101 - Basic obligation of public service.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 2010-01-01 false Basic obligation of public service. 2635.101 Section 2635.101 Administrative...Provisions § 2635.101 Basic obligation of public service. (a) Public service is a public trust. Each...

  4. 49 CFR 371.10 - Duties and obligations of brokers.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ...2010-10-01 false Duties and obligations of brokers. 371.10 Section 371.10 Transportation...FEDERAL MOTOR CARRIER SAFETY REGULATIONS BROKERS OF PROPERTY § 371.10 Duties and obligations of brokers. Where the broker acts on...

  5. 12 CFR 560.42 - State and local government obligations.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ...2010-01-01 2010-01-01 false State and local government obligations. 560.42 Section 560.42 Banks...Federal Savings Associations § 560.42 State and local government obligations. (a) What limitations...

  6. 18 CFR 367.22 - Accounting for asset retirement obligations.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...2010-04-01 2010-04-01 false Accounting for asset retirement obligations...General Instructions § 367.22 Accounting for asset retirement obligations...retirement cost must be stated at the fair value of the asset retirement...

  7. VIEWING RESEARCH PARTICIPATION AS A MORAL OBLIGATION

    PubMed Central

    RENNIE, STUART

    2015-01-01

    A moral paradigm shift has proposed for participation in health-related research. It’s not just a praiseworthy option, some say; it’s a social obligation. Recasting research participation in this way would have global ramifications, however. Who ultimately stands to gain the most from it, and who has the most to lose? PMID:21495516

  8. 33 CFR 137.5 - Disclosure obligations.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 33 Navigation and Navigable Waters 2 2014-07-01 2014-07-01 false Disclosure obligations. 137.5 Section 137.5 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) MARINE POLLUTION FINANCIAL RESPONSIBILITY AND COMPENSATION OIL SPILL LIABILITY: STANDARDS FOR...

  9. 33 CFR 137.5 - Disclosure obligations.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 33 Navigation and Navigable Waters 2 2010-07-01 2010-07-01 false Disclosure obligations. 137.5 Section 137.5 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) MARINE POLLUTION FINANCIAL RESPONSIBILITY AND COMPENSATION OIL SPILL LIABILITY: STANDARDS FOR...

  10. 33 CFR 137.5 - Disclosure obligations.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 33 Navigation and Navigable Waters 2 2011-07-01 2011-07-01 false Disclosure obligations. 137.5 Section 137.5 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) MARINE POLLUTION FINANCIAL RESPONSIBILITY AND COMPENSATION OIL SPILL LIABILITY: STANDARDS FOR...

  11. 33 CFR 137.5 - Disclosure obligations.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 33 Navigation and Navigable Waters 2 2012-07-01 2012-07-01 false Disclosure obligations. 137.5 Section 137.5 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) MARINE POLLUTION FINANCIAL RESPONSIBILITY AND COMPENSATION OIL SPILL LIABILITY: STANDARDS FOR...

  12. 33 CFR 137.5 - Disclosure obligations.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 33 Navigation and Navigable Waters 2 2013-07-01 2013-07-01 false Disclosure obligations. 137.5 Section 137.5 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) MARINE POLLUTION FINANCIAL RESPONSIBILITY AND COMPENSATION OIL SPILL LIABILITY: STANDARDS FOR...

  13. Inability and Obligation in Moral Judgment

    PubMed Central

    Buckwalter, Wesley; Turri, John

    2015-01-01

    It is often thought that judgments about what we ought to do are limited by judgments about what we can do, or that “ought implies can.” We conducted eight experiments to test the link between a range of moral requirements and abilities in ordinary moral evaluations. Moral obligations were repeatedly attributed in tandem with inability, regardless of the type (Experiments 1–3), temporal duration (Experiment 5), or scope (Experiment 6) of inability. This pattern was consistently observed using a variety of moral vocabulary to probe moral judgments and was insensitive to different levels of seriousness for the consequences of inaction (Experiment 4). Judgments about moral obligation were no different for individuals who can or cannot perform physical actions, and these judgments differed from evaluations of a non-moral obligation (Experiment 7). Together these results demonstrate that commonsense morality rejects the “ought implies can” principle for moral requirements, and that judgments about moral obligation are made independently of considerations about ability. By contrast, judgments of blame were highly sensitive to considerations about ability (Experiment 8), which suggests that commonsense morality might accept a “blame implies can” principle. PMID:26296206

  14. The author’s opportunity and obligation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Peer review is a critical component of the scientific method and therefore should be an obligation for everyone who desires to publish their research results in refereed journals. This editorial is written to address a specific problem being encountered by editors of Soil & Tillage Research, but the...

  15. 47 CFR 90.1440 - Reporting obligations.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... Telecommunication FEDERAL COMMUNICATIONS COMMISSION (CONTINUED) SAFETY AND SPECIAL RADIO SERVICES PRIVATE LAND MOBILE RADIO SERVICES 700 MHz Public/Private Partnership § 90.1440 Reporting obligations. (a) The Upper 700 MHz D Block licensee and the Public Safety Broadband Licensee shall jointly file quarterly...

  16. 47 CFR 27.1340 - Reporting obligations.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... COMMUNICATIONS SERVICES 700 MHz Public/Private Partnership § 27.1340 Reporting obligations. (a) The Upper 700 MHz D Block licensee and the Public Safety Broadband Licensee shall jointly file quarterly reports with... requirements of public safety are being met, detailed information on the areas where broadband service has...

  17. 47 CFR 90.1440 - Reporting obligations.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... Telecommunication FEDERAL COMMUNICATIONS COMMISSION (CONTINUED) SAFETY AND SPECIAL RADIO SERVICES PRIVATE LAND MOBILE RADIO SERVICES 700 MHz Public/Private Partnership § 90.1440 Reporting obligations. (a) The Upper 700 MHz D Block licensee and the Public Safety Broadband Licensee shall jointly file quarterly...

  18. 47 CFR 27.1340 - Reporting obligations.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... COMMUNICATIONS SERVICES 700 MHz Public/Private Partnership § 27.1340 Reporting obligations. (a) The Upper 700 MHz D Block licensee and the Public Safety Broadband Licensee shall jointly file quarterly reports with... requirements of public safety are being met, detailed information on the areas where broadband service has...

  19. 47 CFR 27.1340 - Reporting obligations.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... COMMUNICATIONS SERVICES 700 MHz Public/Private Partnership § 27.1340 Reporting obligations. (a) The Upper 700 MHz D Block licensee and the Public Safety Broadband Licensee shall jointly file quarterly reports with... requirements of public safety are being met, detailed information on the areas where broadband service has...

  20. 47 CFR 90.1440 - Reporting obligations.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... Telecommunication FEDERAL COMMUNICATIONS COMMISSION (CONTINUED) SAFETY AND SPECIAL RADIO SERVICES PRIVATE LAND MOBILE RADIO SERVICES 700 MHz Public/Private Partnership § 90.1440 Reporting obligations. (a) The Upper 700 MHz D Block licensee and the Public Safety Broadband Licensee shall jointly file quarterly...

  1. 21 CFR 26.62 - General obligations.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 1 2011-04-01 2011-04-01 false General obligations. 26.62 Section 26.62 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES GENERAL MUTUAL RECOGNITION OF PHARMACEUTICAL GOOD MANUFACTURING PRACTICE REPORTS, MEDICAL DEVICE QUALITY SYSTEM AUDIT...

  2. 21 CFR 26.62 - General obligations.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 1 2010-04-01 2010-04-01 false General obligations. 26.62 Section 26.62 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES GENERAL MUTUAL RECOGNITION OF PHARMACEUTICAL GOOD MANUFACTURING PRACTICE REPORTS, MEDICAL DEVICE QUALITY SYSTEM AUDIT...

  3. 19 CFR 10.412 - Importer obligations.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 19 Customs Duties 1 2014-04-01 2014-04-01 false Importer obligations. 10.412 Section 10.412 Customs Duties U.S. CUSTOMS AND BORDER PROTECTION, DEPARTMENT OF HOMELAND SECURITY; DEPARTMENT OF THE TREASURY ARTICLES CONDITIONALLY FREE, SUBJECT TO A REDUCED RATE, ETC. United States-Chile Free...

  4. 19 CFR 10.412 - Importer obligations.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 19 Customs Duties 1 2010-04-01 2010-04-01 false Importer obligations. 10.412 Section 10.412 Customs Duties U.S. CUSTOMS AND BORDER PROTECTION, DEPARTMENT OF HOMELAND SECURITY; DEPARTMENT OF THE TREASURY ARTICLES CONDITIONALLY FREE, SUBJECT TO A REDUCED RATE, ETC. United States-Chile Free...

  5. 19 CFR 10.412 - Importer obligations.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 19 Customs Duties 1 2011-04-01 2011-04-01 false Importer obligations. 10.412 Section 10.412 Customs Duties U.S. CUSTOMS AND BORDER PROTECTION, DEPARTMENT OF HOMELAND SECURITY; DEPARTMENT OF THE TREASURY ARTICLES CONDITIONALLY FREE, SUBJECT TO A REDUCED RATE, ETC. United States-Chile Free...

  6. 19 CFR 10.412 - Importer obligations.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 19 Customs Duties 1 2013-04-01 2013-04-01 false Importer obligations. 10.412 Section 10.412 Customs Duties U.S. CUSTOMS AND BORDER PROTECTION, DEPARTMENT OF HOMELAND SECURITY; DEPARTMENT OF THE TREASURY ARTICLES CONDITIONALLY FREE, SUBJECT TO A REDUCED RATE, ETC. United States-Chile Free...

  7. 19 CFR 10.412 - Importer obligations.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 19 Customs Duties 1 2012-04-01 2012-04-01 false Importer obligations. 10.412 Section 10.412 Customs Duties U.S. CUSTOMS AND BORDER PROTECTION, DEPARTMENT OF HOMELAND SECURITY; DEPARTMENT OF THE TREASURY ARTICLES CONDITIONALLY FREE, SUBJECT TO A REDUCED RATE, ETC. United States-Chile Free...

  8. 26 CFR 1.454-1 - Obligations issued at discount.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 26 Internal Revenue 6 2011-04-01 2011-04-01 false Obligations issued at discount. 1.454-1 Section... Obligations issued at discount. (a) Certain non-interest-bearing obligations issued at discount—(1) Election... issued at a discount and redeemable for fixed amounts increasing at stated intervals (other than...

  9. 26 CFR 1.454-1 - Obligations issued at discount.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 Internal Revenue 6 2010-04-01 2010-04-01 false Obligations issued at discount. 1.454-1 Section... at discount. (a) Certain non-interest-bearing obligations issued at discount—(1) Election to include... discount and redeemable for fixed amounts increasing at stated intervals (other than an obligation...

  10. 49 CFR 22.17 - Compliance with child support obligations.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 49 Transportation 1 2012-10-01 2012-10-01 false Compliance with child support obligations. 22.17...) Policies Applying to STLP Loans § 22.17 Compliance with child support obligations. Any holder of 50% or... than 60 days delinquent on any obligation to pay child support arising under: (a) An...

  11. 49 CFR 22.17 - Compliance with child support obligations.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 49 Transportation 1 2013-10-01 2013-10-01 false Compliance with child support obligations. 22.17...) Policies Applying to STLP Loans § 22.17 Compliance with child support obligations. Any holder of 50% or... than 60 days delinquent on any obligation to pay child support arising under: (a) An...

  12. 49 CFR 22.17 - Compliance with child support obligations.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 49 Transportation 1 2010-10-01 2010-10-01 false Compliance with child support obligations. 22.17...) Policies Applying to STLP Loans § 22.17 Compliance with child support obligations. Any holder of 50% or... than 60 days delinquent on any obligation to pay child support arising under: (a) An...

  13. 49 CFR 22.17 - Compliance with child support obligations.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 49 Transportation 1 2014-10-01 2014-10-01 false Compliance with child support obligations. 22.17...) Policies Applying to STLP Loans § 22.17 Compliance with child support obligations. Any holder of 50% or... than 60 days delinquent on any obligation to pay child support arising under: (a) An...

  14. Microsporidia Are Natural Intracellular Parasites of the Nematode Caenorhabditis elegans

    PubMed Central

    Troemel, Emily R; Félix, Marie-Anne; Whiteman, Noah K; Barrière, Antoine; Ausubel, Frederick M

    2008-01-01

    For decades the soil nematode Caenorhabditis elegans has been an important model system for biology, but little is known about its natural ecology. Recently, C. elegans has become the focus of studies of innate immunity and several pathogens have been shown to cause lethal intestinal infections in C. elegans. However none of these pathogens has been shown to invade nematode intestinal cells, and no pathogen has been isolated from wild-caught C. elegans. Here we describe an intracellular pathogen isolated from wild-caught C. elegans that we show is a new species of microsporidia. Microsporidia comprise a large class of eukaryotic intracellular parasites that are medically and agriculturally important, but poorly understood. We show that microsporidian infection of the C. elegans intestine proceeds through distinct stages and is transmitted horizontally. Disruption of a conserved cytoskeletal structure in the intestine called the terminal web correlates with the release of microsporidian spores from infected cells, and appears to be part of a novel mechanism by which intracellular pathogens exit from infected cells. Unlike in bacterial intestinal infections, the p38 MAPK and insulin/insulin-like growth factor (IGF) signaling pathways do not appear to play substantial roles in resistance to microsporidian infection in C. elegans. We found microsporidia in multiple wild-caught isolates of Caenorhabditis nematodes from diverse geographic locations. These results indicate that microsporidia are common parasites of C. elegans in the wild. In addition, the interaction between C. elegans and its natural microsporidian parasites provides a system in which to dissect intracellular intestinal infection in vivo and insight into the diversity of pathogenic mechanisms used by intracellular microbes. PMID:19071962

  15. Ecophysiological Characteristics of Obligate Methanotrophic Bacteria and Methane Oxidation In Situ

    NASA Technical Reports Server (NTRS)

    King, Gary M.

    1993-01-01

    Most of the obligate methane-oxidizing bacteria (MOB) described to date are neutrophilic mesophiles that grow optimally in dilute media. Kinetic analyses generally indicate that bacterial methane uptake occurs by transport systems with a K(sub m) greater than l micronM. These and other properties of MOB are inconsistent with characteristics of methane oxidation in situ. The inconsistencies indicate a need for greater attention to the ecophysiological characteristics of isolates and the design of enrichment and isolation schemes which emphasize ecologically relevant parameters (e.g., low temperature, limited and diverse substrate availability, low water potential).

  16. Spatially-resolved intracellular sensing of hydrogen peroxide in living cells.

    PubMed

    Warren, Emilie A K; Netterfield, Tatiana S; Sarkar, Saheli; Kemp, Melissa L; Payne, Christine K

    2015-01-01

    Understanding intracellular redox chemistry requires new tools for the site-specific visualization of intracellular oxidation. We have developed a spatially-resolved intracellular sensor of hydrogen peroxide, HyPer-Tau, for time-resolved imaging in live cells. This sensor consists of a hydrogen peroxide-sensing protein tethered to microtubules. We demonstrate the use of the HyPer-Tau sensor for three applications; dose-dependent response of human cells to exogenous hydrogen peroxide, a model immune response of mouse macrophages to stimulation by bacterial toxin, and a spatially-resolved response to localized delivery of hydrogen peroxide. These results demonstrate that HyPer-Tau can be used as an effective tool for tracking changes in spatially localized intracellular hydrogen peroxide and for future applications in redox signaling. PMID:26585385

  17. Intracellular antibody-mediated immunity and the role of TRIM21.

    PubMed

    McEwan, William A; Mallery, Donna L; Rhodes, David A; Trowsdale, John; James, Leo C

    2011-11-01

    Protection against bacterial and viral pathogens by antibodies has always been thought to end at the cell surface. Once inside the cell, a pathogen was understood to be safe from humoral immunity. However, it has now been found that antibodies can routinely enter cells attached to viral particles and mediate an intracellular immune response. Antibody-coated virions are detected inside the cell by means of an intracellular antibody receptor, TRIM21, which directs their degradation by recruitment of the ubiquitin-proteasome system. In this article we assess how this discovery alters our view of the way in which antibodies neutralise viral infection. We also consider the antiviral function of TRIM21 in the context of its other reported roles in immune signalling and autoimmunity. Finally, we discuss the conceptual implications of intracellular antibody immunity and how it alters our view of the discrete separation of extracellular and intracellular environments. PMID:22006823

  18. Spatially-resolved intracellular sensing of hydrogen peroxide in living cells

    PubMed Central

    Warren, Emilie A. K.; Netterfield, Tatiana S.; Sarkar, Saheli; Kemp, Melissa L.; Payne, Christine K.

    2015-01-01

    Understanding intracellular redox chemistry requires new tools for the site-specific visualization of intracellular oxidation. We have developed a spatially-resolved intracellular sensor of hydrogen peroxide, HyPer-Tau, for time-resolved imaging in live cells. This sensor consists of a hydrogen peroxide-sensing protein tethered to microtubules. We demonstrate the use of the HyPer-Tau sensor for three applications; dose-dependent response of human cells to exogenous hydrogen peroxide, a model immune response of mouse macrophages to stimulation by bacterial toxin, and a spatially-resolved response to localized delivery of hydrogen peroxide. These results demonstrate that HyPer-Tau can be used as an effective tool for tracking changes in spatially localized intracellular hydrogen peroxide and for future applications in redox signaling. PMID:26585385

  19. Interactions between Autophagy and Bacterial Toxins: Targets for Therapy?

    PubMed Central

    Mathieu, Jacques

    2015-01-01

    Autophagy is a physiological process involved in defense mechanisms for clearing intracellular bacteria. The autophagic pathway is finely regulated and bacterial toxins interact with this process in a complex manner. Bacterial toxins also interact significantly with many biochemical processes. Evaluations of the effects of bacterial toxins, such as endotoxins, pore-forming toxins and adenylate cyclases, on autophagy could support the development of new strategies for counteracting bacterial pathogenicity. Treatment strategies could focus on drugs that enhance autophagic processes to improve the clearance of intracellular bacteria. However, further in vivo studies are required to decipher the upregulation of autophagy and potential side effects limiting such approaches. The capacity of autophagy activation strategies to improve the outcome of antibiotic treatment should be investigated in the future. PMID:26248079

  20. Influence of dTMP on the phenotypic appearance and intracellular persistence of Staphylococcus aureus.

    PubMed

    Zander, Johannes; Besier, Silke; Saum, Stephan H; Dehghani, Faramarz; Loitsch, Stefan; Brade, Volker; Wichelhaus, Thomas A

    2008-04-01

    Thymidine-dependent small-colony variants (SCVs) of Staphylococcus aureus are frequently associated with persistent and recurrent infections in cystic fibrosis patients. The phenotypic appearance of S. aureus SCVs or normal-colony variants (NCVs) is postulated to be affected by the intracellular amount of dTMP. This hypothesis was proven by metabolic pathway assays revealing altered intracellular dTMP concentrations, followed by investigation of the associated phenotype. Inhibition of the staphylococcal thymidylate synthase, which generated intracellular dTMP from dUMP, using 5-fluorouracil and co-trimoxazole resulted in an SCV phenotype. Inhibition of a nucleoside transporter, which provided the bacterial cell with extracellular thymidine, caused growth inhibition of SCVs. In turn, reversion of SCVs to NCVs was achieved by supplying extracellular dTMP. High-performance liquid chromatography additionally confirmed the intracellular lack of dTMP in SCVs, in contrast to NCVs. Moreover, the dTMP concentration is postulated to influence the intracellular persistence of S. aureus. Cell culture experiments with cystic fibrosis cells revealed that clinical and co-trimoxazole-induced SCVs with a diminished amount of dTMP showed significantly better intracellular persistence than NCVs. In conclusion, these results show that the dTMP concentration plays a key role in both the phenotypic appearance and the intracellular persistence of S. aureus. PMID:18160477

  1. Direct Measurement of Intracellular Pressure

    PubMed Central

    Petrie, Ryan J.; Koo, Hyun

    2014-01-01

    A method to directly measure the intracellular pressure of adherent, migrating cells is described in the Basic Protocol. This approach is based on the servo-null method where a microelectrode is introduced into the cell to directly measure the physical pressure of the cytoplasm. We also describe the initial calibration of the microelectrode as well as the application of the method to cells migrating inside three-dimensional (3D) extracellular matrix (ECM). PMID:24894836

  2. Intracellular Calcium Channels in Protozoa

    PubMed Central

    Docampo, Roberto; Moreno, Silvia N.J.; Plattner, Helmut

    2014-01-01

    Ca2+-signaling pathways and intracellular Ca2+ channels are present in protozoa. Ancient origin of inositol 1,4,5-trisphosphate receptors (IP3Rs) and other intracellular channels predates the divergence of animals and fungi as evidenced by their presence in the choanoflagellate Monosiga brevicollis, the closest known relative to metazoans. The first protozoan IP3R cloned, from the ciliate Paramecium, displays strong sequence similarity to the rat type 3 IP3R. This ciliate has a large number of IP3- and ryanodine(Ry)-like receptors in 6 subfamilies suggesting the evolutionary adaptation to local requirements for an expanding diversification of vesicle trafficking. IP3Rs have also been functionally characterized in trypanosomatids, where they are essential for growth, differentiation, and establishment of infection. The presence of the mitochondrial calcium uniporter (MCU) in a number of protozoa indicates that mitochondrial regulation of Ca2+ signaling is also an early appearance in evolution, and contributed to the discovery of the molecular nature of this channel in mammalian cells. There is only sequence evidence for the occurrence of two-pore channels (TPCs), transient receptor potential Ca2+ channels (TRPCs) and intracellular mechanosensitive Ca2+-channels in Paramecium and in parasitic protozoa. PMID:24291099

  3. Wholesale service obligation of electric utilities

    SciTech Connect

    Norton, F.L. IV; Spivak, M.R.

    1985-01-01

    The basic concepts of public utility status and utility regulation intertwine the obligation to provide service to the public as reasonably demanded with rate regulation and shielding from competitive interference. While a common law service obligation was not part of the Federal Power Act, the Federal Energy Regulatory Commission has taken the position that service, once commenced, may not be terminated without its approval. This view of Commission authority may not be supported by the legislative history of the Federal Power Act or by judicial precedent. The requirement to serve apart from recognition of a right to serve may result in increased rates in the near term and insufficient capacity, or both, in the long run. A review by the Commission and the courts is examining ways to introduce competition and shift risks from ratepayers to shareholders.

  4. Mechanisms of obligatory intracellular infection with Anaplasma phagocytophilum.

    PubMed

    Rikihisa, Yasuko

    2011-07-01

    Anaplasma phagocytophilum persists in nature by cycling between mammals and ticks. Human infection by the bite of an infected tick leads to a potentially fatal emerging disease called human granulocytic anaplasmosis. A. phagocytophilum is an obligatory intracellular bacterium that replicates inside mammalian granulocytes and the salivary gland and midgut cells of ticks. A. phagocytophilum evolved the remarkable ability to hijack the regulatory system of host cells. A. phagocytophilum alters vesicular traffic to create an intracellular membrane-bound compartment that allows replication in seclusion from lysosomes. The bacterium downregulates or actively inhibits a number of innate immune responses of mammalian host cells, and it upregulates cellular cholesterol uptake to acquire cholesterol for survival. It also upregulates several genes critical for the infection of ticks, and it prolongs tick survival at freezing temperatures. Several host factors that exacerbate infection have been identified, including interleukin-8 (IL-8) and cholesterol. Host factors that overcome infection include IL-12 and gamma interferon (IFN-?). Two bacterial type IV secretion effectors and several bacterial proteins that associate with inclusion membranes have been identified. An understanding of the molecular mechanisms underlying A. phagocytophilum infection will foster the development of creative ideas to prevent or treat this emerging tick-borne disease. PMID:21734244

  5. A detailed view of the intracellular transcriptome of Listeria monocytogenes in murine macrophages using RNA-seq

    PubMed Central

    Schultze, Tilman; Hilker, Rolf; Mannala, Gopala K.; Gentil, Katrin; Weigel, Markus; Farmani, Neda; Windhorst, Anita C.; Goesmann, Alexander; Chakraborty, Trinad; Hain, Torsten

    2015-01-01

    Listeria monocytogenes is a bacterial pathogen and causative agent for the foodborne infection listeriosis, which is mainly a threat for pregnant, elderly, or immunocompromised individuals. Due to its ability to invade and colonize diverse eukaryotic cell types including cells from invertebrates, L. monocytogenes has become a well-established model organism for intracellular growth. Almost 10 years ago, we and others presented the first whole-genome microarray-based intracellular transcriptome of L. monocytogenes. With the advent of newer technologies addressing transcriptomes in greater detail, we revisit this work, and analyze the intracellular transcriptome of L. monocytogenes during growth in murine macrophages using a deep sequencing based approach. We detected 656 differentially expressed genes of which 367 were upregulated during intracellular growth in macrophages compared to extracellular growth in Brain Heart Infusion broth. This study confirmed ?64% of all regulated genes previously identified by microarray analysis. Many of the regulated genes that were detected in the current study involve transporters for various metals, ions as well as complex sugars such as mannose. We also report changes in antisense transcription, especially upregulations during intracellular bacterial survival. A notable finding was the detection of regulatory changes for a subset of temperate A118-like prophage genes, thereby shedding light on the transcriptional profile of this bacteriophage during intracellular growth. In total, our study provides an updated genome-wide view of the transcriptional landscape of L. monocytogenes during intracellular growth and represents a rich resource for future detailed analysis. PMID:26579105

  6. Complete Bacteriophage Transfer in a Bacterial Endosymbiont (Wolbachia) Determined by Targeted

    E-print Network

    Bordenstein, Seth

    Complete Bacteriophage Transfer in a Bacterial Endosymbiont (Wolbachia) Determined by Targeted Bacteriophage flux can cause the majority of genetic diversity in free-living bacteria. This tenet of bacterial the reductive forces of the intracellular lifestyle. To test whether bacteriophages transfer as single genes

  7. In Vitro Models for the Study of the Intracellular Activity of Antibiotics.

    PubMed

    Buyck, Julien M; Lemaire, Sandrine; Seral, Cristina; Anantharajah, Ahalieyah; Peyrusson, Frédéric; Tulkens, Paul M; Van Bambeke, Françoise

    2016-01-01

    Intracellular bacteria are poorly responsive to antibiotic treatment. Pharmacological studies are thus needed to determine which antibiotics are most potent or effective against intracellular bacteria as well as to explore the reasons for poor bacterial responsiveness. An in vitro pharmacodynamic model is described, consisting of (1) phagocytosis of pre-opsonized bacteria by eukaryotic cells; (2) elimination of non-internalized bacteria with gentamicin; (3) incubation of infected cells with antibiotics; and (4) determination of surviving bacteria by viable cell counting and normalization of the counts based on sample protein content. PMID:26468107

  8. Antimycobacterial Efficacy of Andrographis paniculata Leaf Extracts Under Intracellular and Hypoxic Conditions.

    PubMed

    Bhatter, Purva; Gupta, Pooja; Daswani, Poonam; Tetali, Pundarikakshudu; Birdi, Tannaz

    2015-01-01

    The inhibition of the growth of Mycobacterium tuberculosis by the extracts of Andrographis paniculata has been studied using intracellular and axenic hypoxic conditions. The inhibition (confirmed using the gold standard colony forming unit assay) was found to increase with "double stimuli" or higher concentration of the extract. Organic solvent extracts were found to inhibit bacterial growth more than the aqueous extracts under microaerophilic conditions mimicked through axenic and intracellular assays. This could be further explored to evaluate the potential of the plant to be used against nonreplicating/dormant bacilli. PMID:25348959

  9. Pharmacology of intracellular signalling pathways

    PubMed Central

    Nahorski, Stefan R

    2006-01-01

    This article provides a brief and somewhat personalized review of the dramatic developments that have occurred over the last 45 years in our understanding of intracellular signalling pathways associated with G-protein-coupled receptor activation. Signalling via cyclic AMP, the phosphoinositides and Ca2+ is emphasized and these systems have already been revealed as new pharmacological targets. The therapeutic benefits of most of such targets are, however, yet to be realized, but it is certain that the discipline of pharmacology needs to widen its boundaries to meet these challenges in the future. PMID:16402119

  10. What are the limits to the obligations of the nurse?

    PubMed Central

    Edwards, S D

    1996-01-01

    This paper enquires into the nature and the extent of the obligations of nurses. It is argued that nurses appear to be obliged to undertake supererogatory acts if they take clause one of the United Kingdom Central Council for Nursing, Midwifery and Health Visiting (UKCC) Code of Professional Conduct seriously (as, indeed, they are required to do). In the first part of the paper, the nature of nursing obligations is outlined, and then the groups and individuals to whom nurses have obligations are identified. Following a brief discussion of the moral foundation of the nurse's obligations to her/his employer, a common conflict of obligations is identified. Then a distinction is drawn between ordinary and extraordinary moral standards. Appreciation of this is necessary for an understanding of the criterion of what constitutes a supererogatory act. By the definition of supererogatory acts proposed below, it is suggested that actions such as whistleblowing satisfy that definition. PMID:8731534

  11. Bacterial hydrodynamics

    E-print Network

    Eric Lauga

    2015-09-07

    Bacteria predate plants and animals by billions of years. Today, they are the world's smallest cells yet they represent the bulk of the world's biomass, and the main reservoir of nutrients for higher organisms. Most bacteria can move on their own, and the majority of motile bacteria are able to swim in viscous fluids using slender helical appendages called flagella. Low-Reynolds-number hydrodynamics is at the heart of the ability of flagella to generate propulsion at the micron scale. In fact, fluid dynamic forces impact many aspects of bacteriology, ranging from the ability of cells to reorient and search their surroundings to their interactions within mechanically and chemically-complex environments. Using hydrodynamics as an organizing framework, we review the biomechanics of bacterial motility and look ahead to future challenges.

  12. Bacterial hydrodynamics

    E-print Network

    Lauga, Eric

    2015-01-01

    Bacteria predate plants and animals by billions of years. Today, they are the world's smallest cells yet they represent the bulk of the world's biomass, and the main reservoir of nutrients for higher organisms. Most bacteria can move on their own, and the majority of motile bacteria are able to swim in viscous fluids using slender helical appendages called flagella. Low-Reynolds-number hydrodynamics is at the heart of the ability of flagella to generate propulsion at the micron scale. In fact, fluid dynamic forces impact many aspects of bacteriology, ranging from the ability of cells to reorient and search their surroundings to their interactions within mechanically and chemically-complex environments. Using hydrodynamics as an organizing framework, we review the biomechanics of bacterial motility and look ahead to future challenges.

  13. Bacterial Cholangitis.

    PubMed

    Lum, Donald F.; Leung, Joseph W.

    2001-04-01

    The treatment of acute bacterial cholangitis requires broad-spectrum antibiotics to cover against gram-negative aerobic enteric organisms (Escherichia coli, Klebsiella species, and Enterobacter species), gram-positive Enterococcus and anaerobic bacteria (Bacteroides fragilis and Clostridium perfringens). Approximately 20% of patients with acute cholangitis fail to respond to conservative treatment with antibiotic therapy and require urgent biliary decompression, which is the mainstay of therapy. This is best accomplished by endoscopic retrograde cholangiopancreatography (ERCP) and placement of a nasobiliary drainage tube or a large bore (10 F or larger) indwelling plastic stent. Alternative therapy includes percutaneous transhepatic biliary drainage or surgical biliary decompression, but these carry a significantly higher morbidity and mortality. Supportive care includes intravenous fluid hydration to prevent renal failure and close monitoring of vital signs for determination of potential septicemia. PMID:11469972

  14. Real-Time monitoring of intracellular wax ester metabolism

    PubMed Central

    2011-01-01

    Background Wax esters are industrially relevant molecules exploited in several applications of oleochemistry and food industry. At the moment, the production processes mostly rely on chemical synthesis from rather expensive starting materials, and therefore solutions are sought from biotechnology. Bacterial wax esters are attractive alternatives, and especially the wax ester metabolism of Acinetobacter sp. has been extensively studied. However, the lack of suitable tools for rapid and simple monitoring of wax ester metabolism in vivo has partly restricted the screening and analyses of potential hosts and optimal conditions. Results Based on sensitive and specific detection of intracellular long-chain aldehydes, specific intermediates of wax ester synthesis, bacterial luciferase (LuxAB) was exploited in studying the wax ester metabolism in Acinetobacter baylyi ADP1. Luminescence was detected in the cultivation of the strain producing wax esters, and the changes in signal levels could be linked to corresponding cell growth and wax ester synthesis phases. Conclusions The monitoring system showed correlation between wax ester synthesis pattern and luminescent signal. The system shows potential for real-time screening purposes and studies on bacterial wax esters, revealing new aspects to dynamics and role of wax ester metabolism in bacteria. PMID:21961954

  15. Intracellularly Induced Cyclophilins Play an Important Role in Stress Adaptation and Virulence of Brucella abortus

    PubMed Central

    García Fernández, Lucía; DelVecchio, Vito G.; Briones, Gabriel

    2013-01-01

    Brucella is an intracellular bacterial pathogen that causes the worldwide zoonotic disease brucellosis. Brucella virulence relies on its ability to transition to an intracellular lifestyle within host cells. Thus, this pathogen must sense its intracellular localization and then reprogram gene expression for survival within the host cell. A comparative proteomic investigation was performed to identify differentially expressed proteins potentially relevant for Brucella intracellular adaptation. Two proteins identified as cyclophilins (CypA and CypB) were overexpressed in the intracellular environment of the host cell in comparison to laboratory-grown Brucella. To define the potential role of cyclophilins in Brucella virulence, a double-deletion mutant was constructed and its resulting phenotype was characterized. The Brucella abortus ?cypAB mutant displayed increased sensitivity to environmental stressors, such as oxidative stress, pH, and detergents. In addition, the B. abortus ?cypAB mutant strain had a reduced growth rate at lower temperature, a phenotype associated with defective expression of cyclophilins in other microorganisms. The B. abortus ?cypAB mutant also displays reduced virulence in BALB/c mice and defective intracellular survival in HeLa cells. These findings suggest that cyclophilins are important for Brucella virulence and survival in the host cells. PMID:23230297

  16. 7 CFR 1450.105 - Obligations of participant.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ...Agriculture (Continued) COMMODITY CREDIT CORPORATION, DEPARTMENT OF AGRICULTURE LOANS, PURCHASES, AND OTHER OPERATIONS BIOMASS CROP ASSISTANCE PROGRAM (BCAP) Matching Payments § 1450.105 Obligations of participant. (a) All...

  17. Intracellular stresses in patterned cell assemblies.

    PubMed

    Moussus, Michel; der Loughian, Christelle; Fuard, David; Courçon, Marie; Gulino-Debrac, Danielle; Delanoë-Ayari, Hélène; Nicolas, Alice

    2014-04-14

    Confining cells on adhesive patterns allows performing robust, weakly dispersed, statistical analysis. A priori, adhesive patterns could be efficient tools to analyze intracellular cell stress fields, in particular when patterns are used to force the geometry of the cytoskeleton. This tool could then be very helpful in deciphering the relationship between the internal architecture of the cells and the mechanical, intracellular stresses. However, the quantification of the intracellular stresses is still something delicate to perform. Here we first propose a new, very simple and original method to quantify the intracellular stresses, which directly relates the strain the cells impose on the extracellular matrix to the intracellular stress field. This method is used to analyze how confinement influences the intracellular stress field. As a result, we show that the more confined the cells are, the more stressed they will be. The influence of the geometry of the adhesive patterns on the stress patterns is also discussed. PMID:24622969

  18. SERS nanosensors for intracellular redox potential measurements 

    E-print Network

    Auchinvole, Craig Alexander R

    2012-06-22

    Redox regulation and homeostasis are critically important in the regulation of cell function; however, there are significant challenges in quantitatively measuring and monitoring intracellular redox potentials. The work ...

  19. Experimental replacement of an obligate insect symbiont.

    PubMed

    Moran, Nancy A; Yun, Yueli

    2015-02-17

    Symbiosis, the close association of unrelated organisms, has been pivotal in biological diversification. In the obligate symbioses found in many insect hosts, organisms that were once independent are permanently and intimately associated, resulting in expanded ecological capabilities. The primary model for this kind of symbiosis is the association between the bacterium Buchnera and the pea aphid (Acyrthosiphon pisum). A longstanding obstacle to efforts to illuminate genetic changes underlying obligate symbioses has been the inability to experimentally disrupt and reconstitute symbiont-host partnerships. Our experiments show that Buchnera can be experimentally transferred between aphid matrilines and, furthermore, that Buchnera replacement has a massive effect on host fitness. Using a recipient pea aphid matriline containing Buchnera that are heat sensitive because of an allele eliminating the heat shock response of a small chaperone, we reduced native Buchnera through heat exposure and introduced a genetically distinct Buchnera from another matriline, achieving complete replacement and stable inheritance. This transfer disrupted 100 million years (? 1 billion generations) of continuous maternal transmission of Buchnera in its host aphids. Furthermore, aphids with the Buchnera replacement enjoyed a dramatic increase in heat tolerance, directly demonstrating a strong effect of symbiont genotype on host ecology. PMID:25561531

  20. Physiological changes induced in four bacterial strains following oxidative stress.

    PubMed

    Baatout, S; De Boever, P; Mergeay, M

    2006-01-01

    In order to study the behaviour and resistance of bacteria under extreme conditions, physiological changes associated with oxidative stress were monitored using flow cytometry. The study was conducted to assess the maintenance of membrane integrity and potential as well as the esterase activity, the intracellular pH and the production of superoxide anions in four bacterial strains (Ralstonia metallidurans, Escherichia coli, Shewanella oneidensis and Deinococcus radiodurans). The strains were chosen for their potential usefulness in bioremediation. Suspensions of R. metallidurans, E. coli, S. oneidensis and D. radiodurans were submitted to 1 h oxidative stress (H2O2 at various concentrations from 0 to 880 mM). Cell membrane permeability (propidium iodide) and potential (rhodamine-123, 3,3'-dihexyloxacarbocyanine iodide), intracellular esterase activity (fluorescein diacetate), intracellular reactive oxygen species concentration (hydroethidine) and intracellular pH (carboxyflurorescein diacetate succinimidyl ester (5(6)) were monitored to evaluate the physiological state and the overall fitness of individual bacterial cells under oxidative stress. The four bacterial strains exhibited varying sensitivities towards H2O2. However, for all bacterial strains, some physiological damage could already be observed from 13.25 mM H2O2 onwards, in particular with regard to their membrane permeability. Depending on the bacterial strains, moderate to high physiological damage could be observed between 13.25 mM and 220 mM H2O2. Membrane potential, esterase activity, intracellular pH and production of superoxide anion production were considerably modified at high H2O2 concentrations in all four strains. In conclusion, we show that a range of significant physiological alterations occurs when bacteria are challenged with H2O2 and fluorescent staining methods coupled with flow cytometry are useful for monitoring the changes induced not only by oxidative stress but also by other stresses like temperature, radiation, pressure, pH, etc.... PMID:17022450

  1. MavN is a Legionella pneumophila vacuole-associated protein required for efficient iron acquisition during intracellular growth

    PubMed Central

    Isaac, Dervla T.; Laguna, Rita K.; Valtz, Nicole; Isberg, Ralph R.

    2015-01-01

    Iron is essential for the growth and virulence of most intravacuolar pathogens. The mechanisms by which microbes bypass host iron restriction to gain access to this metal across the host vacuolar membrane are poorly characterized. In this work, we identify a unique intracellular iron acquisition strategy used by Legionella pneumophila. The bacterial Icm/Dot (intracellular multiplication/defect in organelle trafficking) type IV secretion system targets the bacterial-derived MavN (more regions allowing vacuolar colocalization N) protein to the surface of the Legionella-containing vacuole where this putative transmembrane protein facilitates intravacuolar iron acquisition. The ?mavN mutant exhibits a transcriptional iron-starvation signature before its growth is arrested during the very early stages of macrophage infection. This intracellular growth defect is rescued only by the addition of excess exogenous iron to the culture medium and not a variety of other metals. Consistent with MavN being a translocated substrate that plays an exclusive role during intracellular growth, the mutant shows no defect for growth in broth culture, even under severe iron-limiting conditions. Putative iron-binding residues within the MavN protein were identified, and point mutations in these residues resulted in defects specific for intracellular growth that are indistinguishable from the ?mavN mutant. This model of a bacterial protein inserting into host membranes to mediate iron transport provides a paradigm for how intravacuolar pathogens can use virulence-associated secretion systems to manipulate and acquire host iron. PMID:26330609

  2. MavN is a Legionella pneumophila vacuole-associated protein required for efficient iron acquisition during intracellular growth.

    PubMed

    Isaac, Dervla T; Laguna, Rita K; Valtz, Nicole; Isberg, Ralph R

    2015-09-15

    Iron is essential for the growth and virulence of most intravacuolar pathogens. The mechanisms by which microbes bypass host iron restriction to gain access to this metal across the host vacuolar membrane are poorly characterized. In this work, we identify a unique intracellular iron acquisition strategy used by Legionella pneumophila. The bacterial Icm/Dot (intracellular multiplication/defect in organelle trafficking) type IV secretion system targets the bacterial-derived MavN (more regions allowing vacuolar colocalization N) protein to the surface of the Legionella-containing vacuole where this putative transmembrane protein facilitates intravacuolar iron acquisition. The ?mavN mutant exhibits a transcriptional iron-starvation signature before its growth is arrested during the very early stages of macrophage infection. This intracellular growth defect is rescued only by the addition of excess exogenous iron to the culture medium and not a variety of other metals. Consistent with MavN being a translocated substrate that plays an exclusive role during intracellular growth, the mutant shows no defect for growth in broth culture, even under severe iron-limiting conditions. Putative iron-binding residues within the MavN protein were identified, and point mutations in these residues resulted in defects specific for intracellular growth that are indistinguishable from the ?mavN mutant. This model of a bacterial protein inserting into host membranes to mediate iron transport provides a paradigm for how intravacuolar pathogens can use virulence-associated secretion systems to manipulate and acquire host iron. PMID:26330609

  3. A lack of parasitic reduction in the obligate parasitic green alga Helicosporidium.

    PubMed

    Pombert, Jean-François; Blouin, Nicolas Achille; Lane, Chris; Boucias, Drion; Keeling, Patrick J

    2014-05-01

    The evolution of an obligate parasitic lifestyle is often associated with genomic reduction, in particular with the loss of functions associated with increasing host-dependence. This is evident in many parasites, but perhaps the most extreme transitions are from free-living autotrophic algae to obligate parasites. The best-known examples of this are the apicomplexans such as Plasmodium, which evolved from algae with red secondary plastids. However, an analogous transition also took place independently in the Helicosporidia, where an obligate parasite of animals with an intracellular infection mechanism evolved from algae with green primary plastids. We characterised the nuclear genome of Helicosporidium to compare its transition to parasitism with that of apicomplexans. The Helicosporidium genome is small and compact, even by comparison with the relatively small genomes of the closely related green algae Chlorella and Coccomyxa, but at the functional level we find almost no evidence for reduction. Nearly all ancestral metabolic functions are retained, with the single major exception of photosynthesis, and even here reduction is not complete. The great majority of genes for light-harvesting complexes, photosystems, and pigment biosynthesis have been lost, but those for other photosynthesis-related functions, such as Calvin cycle, are retained. Rather than loss of whole function categories, the predominant reductive force in the Helicosporidium genome is a contraction of gene family complexity, but even here most losses affect families associated with genome maintenance and expression, not functions associated with host-dependence. Other gene families appear to have expanded in response to parasitism, in particular chitinases, including those predicted to digest the chitinous barriers of the insect host or remodel the cell wall of Helicosporidium. Overall, the Helicosporidium genome presents a fascinating picture of the early stages of a transition from free-living autotroph to parasitic heterotroph where host-independence has been unexpectedly preserved. PMID:24809511

  4. A Lack of Parasitic Reduction in the Obligate Parasitic Green Alga Helicosporidium

    PubMed Central

    Pombert, Jean-François; Blouin, Nicolas Achille; Lane, Chris; Boucias, Drion; Keeling, Patrick J.

    2014-01-01

    The evolution of an obligate parasitic lifestyle is often associated with genomic reduction, in particular with the loss of functions associated with increasing host-dependence. This is evident in many parasites, but perhaps the most extreme transitions are from free-living autotrophic algae to obligate parasites. The best-known examples of this are the apicomplexans such as Plasmodium, which evolved from algae with red secondary plastids. However, an analogous transition also took place independently in the Helicosporidia, where an obligate parasite of animals with an intracellular infection mechanism evolved from algae with green primary plastids. We characterised the nuclear genome of Helicosporidium to compare its transition to parasitism with that of apicomplexans. The Helicosporidium genome is small and compact, even by comparison with the relatively small genomes of the closely related green algae Chlorella and Coccomyxa, but at the functional level we find almost no evidence for reduction. Nearly all ancestral metabolic functions are retained, with the single major exception of photosynthesis, and even here reduction is not complete. The great majority of genes for light-harvesting complexes, photosystems, and pigment biosynthesis have been lost, but those for other photosynthesis-related functions, such as Calvin cycle, are retained. Rather than loss of whole function categories, the predominant reductive force in the Helicosporidium genome is a contraction of gene family complexity, but even here most losses affect families associated with genome maintenance and expression, not functions associated with host-dependence. Other gene families appear to have expanded in response to parasitism, in particular chitinases, including those predicted to digest the chitinous barriers of the insect host or remodel the cell wall of Helicosporidium. Overall, the Helicosporidium genome presents a fascinating picture of the early stages of a transition from free-living autotroph to parasitic heterotroph where host-independence has been unexpectedly preserved. PMID:24809511

  5. The evolution of genomic instability in the obligate endosymbionts of whiteflies.

    PubMed

    Sloan, Daniel B; Moran, Nancy A

    2013-01-01

    Many insects depend on ancient associations with intracellular bacteria to perform essential metabolic functions. These endosymbionts exhibit striking examples of convergence in genome architecture, including a high degree of structural stability that is not typical of their free-living counterparts. However, the recently sequenced genome of the obligate whitefly endosymbiont Portiera revealed features that distinguish it from other ancient insect associates, such as a low gene density and the presence of perfectly duplicated sequences. Here, we report the comparative analysis of Portiera genome sequences both within and between host species. In one whitefly lineage (Bemisia tabaci), we identify large-scale structural polymorphisms in the Portiera genome that exist even within individual insects. This variation is likely mediated by recombination across identical repeats that are maintained by gene conversion. The complete Portiera genome sequence from a distantly related whitefly host (Trialeurodes vaporarium) confirms a history of extensive genome rearrangement in this ancient endosymbiont. Using gene-order-based phylogenetic analysis, we show that the majority of rearrangements have occurred in the B. tabaci lineage, coinciding with an increase in the rate of nucleotide substitutions, a proliferation of short tandem repeats (microsatellites) in intergenic regions, and the loss of many widely conserved genes involved in DNA replication, recombination, and repair. These results indicate that the loss of recombinational machinery is unlikely to be the cause of the extreme structural conservation that is generally observed in obligate endosymbiont genomes and that large, repetitive intergenic regions are an important substrate for genomic rearrangements. PMID:23542079

  6. Bacterial vaginosis.

    PubMed Central

    Spiegel, C A

    1991-01-01

    Bacterial vaginosis (BV) is the most common of the vaginitides affecting women of reproductive age. It appears to be due to an alteration in the vaginal ecology by which Lactobacillus spp., the predominant organisms in the healthy vagina, are replaced by a mixed flora including Prevotella bivia, Prevotella disiens, Porphyromonas spp., Mobiluncus spp., and Peptostreptococcus spp. All of these organisms except Mobiluncus spp. are also members of the endogenous vaginal flora. While evidence from treatment trials does not support the notion that BV is sexually transmitted, recent studies have shown an increased risk associated with multiple sexual partners. It has also been suggested that the pathogenesis of BV may be similar to that of urinary tract infections, with the rectum serving as a reservoir for some BV-associated flora. The organisms associated with BV have also been recognized as agents of female upper genital tract infection, including pelvic inflammatory disease, and the syndrome BV has been associated with adverse outcome of pregnancy, including premature rupture of membranes, chorioamnionitis, and fetal loss; postpartum endometritis; cuff cellulitis; and urinary tract infections. The mechanisms by which the BV-associated flora causes the signs of BV are not well understood, but a role for H2O2-producing Lactobacillus spp. in protecting against colonization by catalase-negative anaerobic bacteria has been recognized. These and other aspects of BV are reviewed. PMID:1747864

  7. Magnetic microbes: Bacterial magnetite biomineralization

    DOE PAGESBeta

    Prozorov, Tanya

    2015-09-14

    Magnetotactic bacteria are a diverse group of prokaryotes with the ability to orient and migrate along the magnetic field lines in search for a preferred oxygen concentration in chemically stratified water columns and sediments. These microorganisms produce magnetosomes, the intracellular nanometer-sized magnetic crystals surrounded by a phospholipid bilayer membrane, typically organized in chains. Magnetosomes have nearly perfect crystal structures with narrow size distribution and species-specific morphologies, leading to well-defined magnetic properties. As a result, the magnetite biomineralization in these organisms is of fundamental interest to diverse disciplines, from biotechnology to astrobiology. As a result, this article highlights recent advances inmore »the understanding of the bacterial magnetite biomineralization.« less

  8. 29 CFR 37.21 - How long will the recipient's obligation under the assurance last, and how broad is the obligation?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 29 Labor 1 2010-07-01 2010-07-01 true How long will the recipient's obligation under the assurance last, and how broad is the obligation? 37.21 Section 37.21 Labor Office of the Secretary of Labor...'s obligation under the assurance last, and how broad is the obligation? (a) Where the WIA Title...

  9. 12 CFR 614.4358 - Computation of obligations.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 12 Banks and Banking 6 2011-01-01 2011-01-01 false Computation of obligations. 614.4358 Section 614.4358 Banks and Banking FARM CREDIT ADMINISTRATION FARM CREDIT SYSTEM LOAN POLICIES AND OPERATIONS Lending and Leasing Limits § 614.4358 Computation of obligations. (a) Inclusions. The computation of...

  10. 18 CFR 35.18 - Asset retirement obligations.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 18 Conservation of Power and Water Resources 1 2013-04-01 2013-04-01 false Asset retirement obligations. 35.18 Section 35.18 Conservation of Power and Water Resources FEDERAL ENERGY REGULATORY... Other Filing Requirements § 35.18 Asset retirement obligations. (a) A public utility that files a...

  11. 18 CFR 35.18 - Asset retirement obligations.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 18 Conservation of Power and Water Resources 1 2014-04-01 2014-04-01 false Asset retirement obligations. 35.18 Section 35.18 Conservation of Power and Water Resources FEDERAL ENERGY REGULATORY... Other Filing Requirements § 35.18 Asset retirement obligations. (a) A public utility that files a...

  12. 18 CFR 35.18 - Asset retirement obligations.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 18 Conservation of Power and Water Resources 1 2011-04-01 2011-04-01 false Asset retirement obligations. 35.18 Section 35.18 Conservation of Power and Water Resources FEDERAL ENERGY REGULATORY... Other Filing Requirements § 35.18 Asset retirement obligations. (a) A public utility that files a...

  13. 18 CFR 35.18 - Asset retirement obligations.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 18 Conservation of Power and Water Resources 1 2012-04-01 2012-04-01 false Asset retirement obligations. 35.18 Section 35.18 Conservation of Power and Water Resources FEDERAL ENERGY REGULATORY... Other Filing Requirements § 35.18 Asset retirement obligations. (a) A public utility that files a...

  14. 12 CFR 612.2270 - Purchase of System obligations.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 12 Banks and Banking 6 2010-01-01 2010-01-01 false Purchase of System obligations. 612.2270... REFERRAL OF KNOWN OR SUSPECTED CRIMINAL VIOLATIONS Standards of Conduct § 612.2270 Purchase of System... Funding Corporation, may only purchase joint, consolidated, or Systemwide obligations that are: (1)...

  15. 12 CFR 612.2270 - Purchase of System obligations.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 12 Banks and Banking 6 2011-01-01 2011-01-01 false Purchase of System obligations. 612.2270... REFERRAL OF KNOWN OR SUSPECTED CRIMINAL VIOLATIONS Standards of Conduct § 612.2270 Purchase of System... Funding Corporation, may only purchase joint, consolidated, or Systemwide obligations that are: (1)...

  16. 7 CFR 984.54 - Establishment of obligation.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... Agreements and Orders; Fruits, Vegetables, Nuts), DEPARTMENT OF AGRICULTURE WALNUTS GROWN IN CALIFORNIA Order Regulating Handling Reserve Walnuts § 984.54 Establishment of obligation. (a) Reserve obligation. Whenever... kernelweight of certified merchantable walnuts equal to a quantity derived by the application of the...

  17. 7 CFR 984.54 - Establishment of obligation.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... AGREEMENTS AND ORDERS; FRUITS, VEGETABLES, NUTS), DEPARTMENT OF AGRICULTURE WALNUTS GROWN IN CALIFORNIA Order Regulating Handling Reserve Walnuts § 984.54 Establishment of obligation. (a) Reserve obligation. Whenever... kernelweight of certified merchantable walnuts equal to a quantity derived by the application of the...

  18. 7 CFR 984.54 - Establishment of obligation.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... AGREEMENTS AND ORDERS; FRUITS, VEGETABLES, NUTS), DEPARTMENT OF AGRICULTURE WALNUTS GROWN IN CALIFORNIA Order Regulating Handling Reserve Walnuts § 984.54 Establishment of obligation. (a) Reserve obligation. Whenever... kernelweight of certified merchantable walnuts equal to a quantity derived by the application of the...

  19. 7 CFR 984.54 - Establishment of obligation.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... Agreements and Orders; Fruits, Vegetables, Nuts), DEPARTMENT OF AGRICULTURE WALNUTS GROWN IN CALIFORNIA Order Regulating Handling Reserve Walnuts § 984.54 Establishment of obligation. (a) Reserve obligation. Whenever... kernelweight of certified merchantable walnuts equal to a quantity derived by the application of the...

  20. 7 CFR 984.54 - Establishment of obligation.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... Agreements and Orders; Fruits, Vegetables, Nuts), DEPARTMENT OF AGRICULTURE WALNUTS GROWN IN CALIFORNIA Order Regulating Handling Reserve Walnuts § 984.54 Establishment of obligation. (a) Reserve obligation. Whenever... kernelweight of certified merchantable walnuts equal to a quantity derived by the application of the...

  1. 47 CFR 211.7 - Obligation of carriers.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 47 Telecommunication 5 2012-10-01 2012-10-01 false Obligation of carriers. 211.7 Section 211.7 Telecommunication OFFICE OF SCIENCE AND TECHNOLOGY POLICY AND NATIONAL SECURITY COUNCIL EMERGENCY RESTORATION PRIORITY PROCEDURES FOR TELECOMMUNICATIONS SERVICES § 211.7 Obligation of carriers. (a) During...

  2. 22 CFR 221.15 - Fiscal Agent obligations.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 22 Foreign Relations 1 2013-04-01 2013-04-01 false Fiscal Agent obligations. 221.15 Section 221.15 Foreign Relations AGENCY FOR INTERNATIONAL DEVELOPMENT ISRAEL LOAN GUARANTEE STANDARD TERMS AND CONDITIONS The Guarantee § 221.15 Fiscal Agent obligations. Failure of the Fiscal Agent to perform any of...

  3. 22 CFR 221.15 - Fiscal Agent obligations.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 22 Foreign Relations 1 2014-04-01 2014-04-01 false Fiscal Agent obligations. 221.15 Section 221.15 Foreign Relations AGENCY FOR INTERNATIONAL DEVELOPMENT ISRAEL LOAN GUARANTEE STANDARD TERMS AND CONDITIONS The Guarantee § 221.15 Fiscal Agent obligations. Failure of the Fiscal Agent to perform any of...

  4. 22 CFR 221.15 - Fiscal Agent obligations.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 22 Foreign Relations 1 2012-04-01 2012-04-01 false Fiscal Agent obligations. 221.15 Section 221.15 Foreign Relations AGENCY FOR INTERNATIONAL DEVELOPMENT ISRAEL LOAN GUARANTEE STANDARD TERMS AND CONDITIONS The Guarantee § 221.15 Fiscal Agent obligations. Failure of the Fiscal Agent to perform any of...

  5. 22 CFR 221.15 - Fiscal Agent obligations.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 22 Foreign Relations 1 2011-04-01 2011-04-01 false Fiscal Agent obligations. 221.15 Section 221.15 Foreign Relations AGENCY FOR INTERNATIONAL DEVELOPMENT ISRAEL LOAN GUARANTEE STANDARD TERMS AND CONDITIONS The Guarantee § 221.15 Fiscal Agent obligations. Failure of the Fiscal Agent to perform any of...

  6. 22 CFR 221.15 - Fiscal Agent obligations.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 22 Foreign Relations 1 2010-04-01 2010-04-01 false Fiscal Agent obligations. 221.15 Section 221.15 Foreign Relations AGENCY FOR INTERNATIONAL DEVELOPMENT ISRAEL LOAN GUARANTEE STANDARD TERMS AND CONDITIONS The Guarantee § 221.15 Fiscal Agent obligations. Failure of the Fiscal Agent to perform any of...

  7. 18 CFR 37.8 - Obligations of OASIS users.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ...2013-04-01 2013-04-01 false Obligations of OASIS users. 37.8 Section 37.8 Conservation...SAME-TIME INFORMATION SYSTEMS § 37.8 Obligations of OASIS users. Each OASIS user must notify the Responsible Party one...

  8. 18 CFR 37.8 - Obligations of OASIS users.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...2011-04-01 2011-04-01 false Obligations of OASIS users. 37.8 Section 37.8 Conservation...SAME-TIME INFORMATION SYSTEMS § 37.8 Obligations of OASIS users. Each OASIS user must notify the Responsible Party one...

  9. 18 CFR 37.8 - Obligations of OASIS users.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ...2012-04-01 2012-04-01 false Obligations of OASIS users. 37.8 Section 37.8 Conservation...SAME-TIME INFORMATION SYSTEMS § 37.8 Obligations of OASIS users. Each OASIS user must notify the Responsible Party one...

  10. 18 CFR 37.8 - Obligations of OASIS users.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...2010-04-01 2010-04-01 false Obligations of OASIS users. 37.8 Section 37.8 Conservation...SAME-TIME INFORMATION SYSTEMS § 37.8 Obligations of OASIS users. Each OASIS user must notify the Responsible Party one...

  11. 18 CFR 37.8 - Obligations of OASIS users.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ...2014-04-01 2014-04-01 false Obligations of OASIS users. 37.8 Section 37.8 Conservation...SAME-TIME INFORMATION SYSTEMS § 37.8 Obligations of OASIS users. Each OASIS user must notify the Responsible Party one...

  12. 18 CFR 37.8 - Obligations of OASIS users.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 18 Conservation of Power and Water Resources 1 2013-04-01 2013-04-01 false Obligations of OASIS... Obligations of OASIS users. Each OASIS user must notify the Responsible Party one month in advance of initiating a significant amount of automated queries. The OASIS user must also notify the Responsible...

  13. 18 CFR 37.8 - Obligations of OASIS users.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 18 Conservation of Power and Water Resources 1 2014-04-01 2014-04-01 false Obligations of OASIS... Obligations of OASIS users. Each OASIS user must notify the Responsible Party one month in advance of initiating a significant amount of automated queries. The OASIS user must also notify the Responsible...

  14. 18 CFR 37.8 - Obligations of OASIS users.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 18 Conservation of Power and Water Resources 1 2011-04-01 2011-04-01 false Obligations of OASIS... Obligations of OASIS users. Each OASIS user must notify the Responsible Party one month in advance of initiating a significant amount of automated queries. The OASIS user must also notify the Responsible...

  15. 18 CFR 37.8 - Obligations of OASIS users.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 18 Conservation of Power and Water Resources 1 2010-04-01 2010-04-01 false Obligations of OASIS... Obligations of OASIS users. Each OASIS user must notify the Responsible Party one month in advance of initiating a significant amount of automated queries. The OASIS user must also notify the Responsible...

  16. 18 CFR 37.8 - Obligations of OASIS users.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 18 Conservation of Power and Water Resources 1 2012-04-01 2012-04-01 false Obligations of OASIS... Obligations of OASIS users. Each OASIS user must notify the Responsible Party one month in advance of initiating a significant amount of automated queries. The OASIS user must also notify the Responsible...

  17. 7 CFR 783.7 - Obligations of a participant.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 7 2010-01-01 2010-01-01 false Obligations of a participant. 783.7 Section 783.7 Agriculture Regulations of the Department of Agriculture (Continued) FARM SERVICE AGENCY, DEPARTMENT OF AGRICULTURE SPECIAL PROGRAMS TREE ASSISTANCE PROGRAM § 783.7 Obligations of a participant. (a)...

  18. 5 CFR 2635.101 - Basic obligation of public service.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 5 Administrative Personnel 3 2010-01-01 2010-01-01 false Basic obligation of public service. 2635.101 Section 2635.101 Administrative Personnel OFFICE OF GOVERNMENT ETHICS GOVERNMENT ETHICS STANDARDS OF ETHICAL CONDUCT FOR EMPLOYEES OF THE EXECUTIVE BRANCH General Provisions § 2635.101 Basic obligation of public service. (a) Public service is...

  19. 34 CFR 75.707 - When obligations are made.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ...2010-07-01 false When obligations are made. 75.707 Section 75.707 Education...Education DIRECT GRANT PROGRAMS What Are the Administrative Responsibilities of a...Responsibilities § 75.707 When obligations are made. The following table shows...

  20. 34 CFR 76.707 - When obligations are made.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ...2010-07-01 false When obligations are made. 76.707 Section 76.707 Education...Education STATE-ADMINISTERED PROGRAMS What Are the Administrative Responsibilities of...Responsibilities § 76.707 When obligations are made. The following table shows...

  1. 47 CFR 76.309 - Customer service obligations.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 47 Telecommunication 4 2014-10-01 2014-10-01 false Customer service obligations. 76.309 Section 76.309 Telecommunication FEDERAL COMMUNICATIONS COMMISSION (CONTINUED) BROADCAST RADIO SERVICES MULTICHANNEL VIDEO AND CABLE TELEVISION SERVICE General Operating Requirements § 76.309 Customer service obligations. (a) A cable franchise authority...

  2. 29 CFR 4043.20 - Post-Event filing obligation.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 29 Labor 9 2012-07-01 2012-07-01 false Post-Event filing obligation. 4043.20 Section 4043.20 Labor... EVENTS AND CERTAIN OTHER NOTIFICATION REQUIREMENTS Post-Event Notice of Reportable Events § 4043.20 Post-Event filing obligation. The plan administrator and each contributing sponsor of a plan for which...

  3. 29 CFR 4043.20 - Post-Event filing obligation.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 29 Labor 9 2014-07-01 2014-07-01 false Post-Event filing obligation. 4043.20 Section 4043.20 Labor... EVENTS AND CERTAIN OTHER NOTIFICATION REQUIREMENTS Post-Event Notice of Reportable Events § 4043.20 Post-Event filing obligation. The plan administrator and each contributing sponsor of a plan for which...

  4. 29 CFR 4043.20 - Post-Event filing obligation.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 29 Labor 9 2013-07-01 2013-07-01 false Post-Event filing obligation. 4043.20 Section 4043.20 Labor... EVENTS AND CERTAIN OTHER NOTIFICATION REQUIREMENTS Post-Event Notice of Reportable Events § 4043.20 Post-Event filing obligation. The plan administrator and each contributing sponsor of a plan for which...

  5. 29 CFR 4043.20 - Post-Event filing obligation.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 29 Labor 9 2011-07-01 2011-07-01 false Post-Event filing obligation. 4043.20 Section 4043.20 Labor... EVENTS AND CERTAIN OTHER NOTIFICATION REQUIREMENTS Post-Event Notice of Reportable Events § 4043.20 Post-Event filing obligation. The plan administrator and each contributing sponsor of a plan for which...

  6. 29 CFR 4043.20 - Post-Event filing obligation.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 29 Labor 9 2010-07-01 2010-07-01 false Post-Event filing obligation. 4043.20 Section 4043.20 Labor... EVENTS AND CERTAIN OTHER NOTIFICATION REQUIREMENTS Post-Event Notice of Reportable Events § 4043.20 Post-Event filing obligation. The plan administrator and each contributing sponsor of a plan for which...

  7. 34 CFR 686.40 - Documenting the service obligation.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... (TEACH) GRANT PROGRAM Service and Repayment Obligations § 686.40 Documenting the service obligation. (a... in a program of study for which a TEACH Grant was received, the grant recipient must confirm to the... period being certified in any of the high-need fields of mathematics, science, a foreign...

  8. Parental Beliefs about Nonresident Fathers' Obligations and Rights

    ERIC Educational Resources Information Center

    Lin, I-Fen; McLanahan, Sara S.

    2007-01-01

    We examine whether parents rely on principles of equity or equality in making judgments about nonresident fathers' obligations and rights. The data are taken from the first wave of the Fragile Families and Child Wellbeing Study. The analysis sample includes 4,304 new mothers and 3,414 new fathers. Results indicate that fathers perceive obligations

  9. 43 CFR 3162.2-1 - Drilling and producing obligations.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 43 Public Lands: Interior 2 2012-10-01 2012-10-01 false Drilling and producing obligations. 3162.2... Requirements for Operating Rights Owners and Operators § 3162.2-1 Drilling and producing obligations. (a) The operator, at its election, may drill and produce other wells in conformity with any system of well...

  10. 43 CFR 3162.2-1 - Drilling and producing obligations.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 43 Public Lands: Interior 2 2011-10-01 2011-10-01 false Drilling and producing obligations. 3162.2... Requirements for Operating Rights Owners and Operators § 3162.2-1 Drilling and producing obligations. (a) The operator, at its election, may drill and produce other wells in conformity with any system of well...

  11. 43 CFR 3162.2-1 - Drilling and producing obligations.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 43 Public Lands: Interior 2 2014-10-01 2014-10-01 false Drilling and producing obligations. 3162.2... Requirements for Operating Rights Owners and Operators § 3162.2-1 Drilling and producing obligations. (a) The operator, at its election, may drill and produce other wells in conformity with any system of well...

  12. 43 CFR 3162.2-1 - Drilling and producing obligations.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 43 Public Lands: Interior 2 2013-10-01 2013-10-01 false Drilling and producing obligations. 3162.2... Requirements for Operating Rights Owners and Operators § 3162.2-1 Drilling and producing obligations. (a) The operator, at its election, may drill and produce other wells in conformity with any system of well...

  13. 29 CFR 4.146 - Contract obligations after award, generally.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 29 Labor 1 2010-07-01 2010-07-01 true Contract obligations after award, generally. 4.146 Section 4.146 Labor Office of the Secretary of Labor LABOR STANDARDS FOR FEDERAL SERVICE CONTRACTS Application of the McNamara-O'Hara Service Contract Act Period of Coverage § 4.146 Contract obligations...

  14. 26 CFR 1.103-16 - Obligations of certain volunteer fire departments.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...2011-04-01 false Obligations of certain volunteer fire departments. 1.103-16...103-16 Obligations of certain volunteer fire departments. (a) General rule. An obligation of a volunteer fire department issued after...

  15. 26 CFR 1.103-16 - Obligations of certain volunteer fire departments.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...2010-04-01 false Obligations of certain volunteer fire departments. 1.103-16...103-16 Obligations of certain volunteer fire departments. (a) General rule. An obligation of a volunteer fire department issued after...

  16. 26 CFR 1.1504-4 - Treatment of warrants, options, convertible obligations, and other similar interests.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...2011-04-01 false Treatment of warrants, options, convertible obligations, and other...1.1504-4 Treatment of warrants, options, convertible obligations, and other...regarding the circumstances in which warrants, options, obligations convertible into...

  17. 26 CFR 1.1504-4 - Treatment of warrants, options, convertible obligations, and other similar interests.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...2010-04-01 false Treatment of warrants, options, convertible obligations, and other...1.1504-4 Treatment of warrants, options, convertible obligations, and other...regarding the circumstances in which warrants, options, obligations convertible into...

  18. 12 CFR 1510.5 - How does the Funding Corporation make interest payments on its obligations?

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ...Funding Corporation make interest payments on its obligations? 1510.5 Section 1510...Funding Corporation make interest payments on its obligations? (a) The Funding Corporation...Corporation must pay the interest due on its obligations with funds it obtains...

  19. 12 CFR 1510.5 - How does the Funding Corporation make interest payments on its obligations?

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ...Funding Corporation make interest payments on its obligations? 1510.5 Section 1510...Funding Corporation make interest payments on its obligations? (a) The Funding Corporation...Corporation must pay the interest due on its obligations with funds it obtains...

  20. 12 CFR 1270.18 - Additional requirements; notice of attachment for Book-entry consolidated obligations.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ...requirements; notice of attachment for Book-entry consolidated obligations. 1270...FEDERAL HOME LOAN BANKS LIABILITIES Book-Entry Procedure for Consolidated Obligations...requirements; notice of attachment for Book-entry consolidated obligations....

  1. 12 CFR 1270.18 - Additional requirements; notice of attachment for Book-entry consolidated obligations.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ...requirements; notice of attachment for Book-entry consolidated obligations. 1270...FEDERAL HOME LOAN BANKS LIABILITIES Book-Entry Procedure for Consolidated Obligations...requirements; notice of attachment for Book-entry consolidated obligations....

  2. 12 CFR 1270.18 - Additional requirements; notice of attachment for Book-entry consolidated obligations.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ...requirements; notice of attachment for Book-entry consolidated obligations. 1270...FEDERAL HOME LOAN BANKS LIABILITIES Book-Entry Procedure for Consolidated Obligations...requirements; notice of attachment for Book-entry consolidated obligations....

  3. 31 CFR 285.8 - Offset of tax refund payments to collect state income tax obligations.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ...State income tax obligation means State income tax obligations as determined under State law. For purposes of this section, State income tax obligation includes any...the overpayment. (b) General rule. (1) FMS will collect...

  4. 31 CFR 225.9 - Return of Government obligations to obligor.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ...2010-07-01 false Return of Government obligations to obligor. 225... ACCEPTANCE OF BONDS SECURED BY GOVERNMENT OBLIGATIONS IN LIEU OF BONDS WITH SURETIES § 225.9 Return of Government obligations to obligor....

  5. 31 CFR 225.9 - Return of Government obligations to obligor.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ...2011-07-01 false Return of Government obligations to obligor. 225... ACCEPTANCE OF BONDS SECURED BY GOVERNMENT OBLIGATIONS IN LIEU OF BONDS WITH SURETIES § 225.9 Return of Government obligations to obligor....

  6. 12 CFR 987.8 - Additional requirements; notice of attachment for Book-entry consolidated obligations.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ...requirements; notice of attachment for Book-entry consolidated obligations. 987...HOUSING FINANCE BOARD OFFICE OF FINANCE BOOK-ENTRY PROCEDURE FOR CONSOLIDATED OBLIGATIONS...requirements; notice of attachment for Book-entry consolidated obligations....

  7. 12 CFR 987.8 - Additional requirements; notice of attachment for Book-entry consolidated obligations.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ...requirements; notice of attachment for Book-entry consolidated obligations. 987...HOUSING FINANCE BOARD OFFICE OF FINANCE BOOK-ENTRY PROCEDURE FOR CONSOLIDATED OBLIGATIONS...requirements; notice of attachment for Book-entry consolidated obligations....

  8. The origins of cooperative bacterial communities.

    PubMed

    Sachs, J L; Hollowell, A C

    2012-01-01

    Bacteria live in complex multispecies communities. Intimately interacting bacterial cells are ubiquitous on biological and mineral surfaces in all habitats. Molecular and cellular biologists have unraveled some key mechanisms that modulate bacterial interactions, but the ecology and evolution of these associations remain poorly understood. One debate has focused on the relative importance of cooperation among cells in bacterial communities. Some researchers suggest that communication and cooperation, both within and among bacterial species, have produced emergent properties that give such groups a selective advantage. Evolutionary biologists have countered that the appearance of group-level traits should be viewed with caution, as natural selection almost invariably favors selfishness. A recent theory by Morris, Lenski, and Zinser, called the Black Queen Hypothesis, gives a new perspective on this debate (J. J. Morris, R. E. Lenski, and E. R. Zinser, mBio 3(2):e00036-12, 2012). These authors present a model that reshapes a decades-old idea: cooperation among species can be automatic and based upon purely selfish traits. Moreover, this hypothesis stands in contrast to the Red Queen Hypothesis, which states that species are in constant evolutionary conflict. Two assumptions serve as the core of the Black Queen model. First, bacterial functions are often leaky, such that cells unavoidably produce resources that benefit others. Second, the receivers of such by-products will tend to delete their own costly pathways for those products, thus building dependency into the interactions. Although not explicitly required in their model, an emergent prediction is that the initiation of such dependency can favor the spread of more obligate coevolved partnerships. This new paradigm suggests that bacteria might often form interdependent cooperative interactions in communities and moreover that bacterial cooperation should leave a clear genomic signature via complementary loss of shared diffusible functions. PMID:22532558

  9. Intracellular sensing of complement C3 activates cell autonomous immunity

    PubMed Central

    Tam, Jerry C.H.; Bidgood, Susanna R.; McEwan, William A.; James, Leo C.

    2014-01-01

    Pathogens traverse multiple barriers during infection including cell membranes. Here we show that during this transition pathogens carry covalently attached complement C3 into the cell, triggering immediate signalling and effector responses. Sensing of C3 in the cytosol activates MAVS-dependent signalling cascades and induces proinflammatory cytokine secretion. C3 also flags viruses for rapid proteasomal degradation, thereby preventing their replication. This system can detect both viral and bacterial pathogens but is antagonized by enteroviruses, such as rhinovirus and poliovirus, which cleave C3 using their 3C protease. The antiviral Rupintrivir inhibits 3C protease and prevents C3 cleavage, rendering enteroviruses susceptible to intracellular complement sensing. Thus, complement C3 allows cells to detect and disable pathogens that have invaded the cytosol. PMID:25190799

  10. Bacterial communities of two parthenogenetic aphid species cocolonizing two host plants across the Hawaiian Islands.

    PubMed

    Jones, Ryan T; Bressan, Alberto; Greenwell, April M; Fierer, Noah

    2011-12-01

    Aphids (Hemiptera: Aphididae) have been the focus of several studies with respect to their interactions with inherited symbionts, but bacterial communities of most aphid species are still poorly characterized. In this research, we used bar-coded pyrosequencing to characterize bacterial communities in aphids. Specifically, we examined the diversity of bacteria in two obligately parthenogenetic aphid species (the melon aphid, Aphis gossypii, and the cardamom aphid, Pentalonia caladii) cocolonizing two plant species (taro, Colocasia esculenta, and ginger, Alpinia purpurata) across four Hawaiian Islands (Hawaii, Kauai, Maui, and Oahu). Results from this study revealed that heritable symbionts dominated the bacterial communities for both aphid species. The bacterial communities differed significantly between the two species, and A. gossypii harbored a more diverse bacterial community than P. caladii. The bacterial communities also differed across aphid populations sampled from the different islands; however, communities did not differ between aphids collected from the two host plants. PMID:21965398

  11. Biophysical Characterization of Chlamydia trachomatis CT584 Supports Its Potential Role as a Type III Secretion Needle Tip Protein

    E-print Network

    Markham, Aaron P.; Jaafar, Zane A.; Kemege, Kyle Evan; Middaugh, C. Russell; Hefty, P. Scott

    2009-11-01

    Chlamydia are obligate intracellular bacterial pathogens that cause a variety of diseases. Likemany Gram-negative bacteria, they employ type III secretion systems (T3SS) for invasion, establishing and maintaining their unique intracellular niche...

  12. Settling down: the genome of Serratia symbiotica from the aphid Cinara tujafilina zooms in on the process of accommodation to a cooperative intracellular life.

    PubMed

    Manzano-Marín, Alejandro; Latorre, Amparo

    2014-07-01

    Particularly interesting cases of mutualistic endosymbioses come from the establishment of co-obligate associations of more than one species of endosymbiotic bacteria. Throughout symbiotic accommodation from a free-living bacterium, passing through a facultative stage and ending as an obligate intracellular one, the symbiont experiences massive genomic losses and phenotypic adjustments. Here, we scrutinized the changes in the coevolution of Serratia symbiotica and Buchnera aphidicola endosymbionts in aphids, paying particular attention to the transformations undergone by S. symbiotica to become an obligate endosymbiont. Although it is already known that S. symbiotica is facultative in Acyrthosiphon pisum, in Cinara cedri it has established a co-obligate endosymbiotic consortium along with B. aphidicola to fulfill the aphid's nutritional requirements. The state of this association in C. tujafilina, an aphid belonging to the same subfamily (Lachninae) that C. cedri, remained unknown. Here, we report the genome of S. symbiotica strain SCt-VLC from the aphid C. tujafilina. While being phylogenetically and genomically very closely related to the facultative endosymbiont S. symbiotica from the aphid A. pisum, it shows a variety of metabolic, genetic, and architectural features, which point toward this endosymbiont being one step closer to an obligate intracellular one. We also describe in depth the process of genome rearrangements suffered by S. symbiotica and the role mobile elements play in gene inactivations. Finally, we postulate the supply to the host of the essential riboflavin (vitamin B2) as key to the establishment of S. symbiotica as a co-obligate endosymbiont in the aphids belonging to the subfamily Lachninane. PMID:24951564

  13. Intracellular signalling during neutrophil recruitment

    PubMed Central

    Mócsai, Attila; Walzog, Barbara; Lowell, Clifford A.

    2015-01-01

    Recruitment of leucocytes such as neutrophils to the extravascular space is a critical step of the inflammation process and plays a major role in the development of various diseases including several cardiovascular diseases. Neutrophils themselves play a very active role in that process by sensing their environment and responding to the extracellular cues by adhesion and de-adhesion, cellular shape changes, chemotactic migration, and other effector functions of cell activation. Those responses are co-ordinated by a number of cell surface receptors and their complex intracellular signal transduction pathways. Here, we review neutrophil signal transduction processes critical for recruitment to the site of inflammation. The two key requirements for neutrophil recruitment are the establishment of appropriate chemoattractant gradients and the intrinsic ability of the cells to migrate along those gradients. We will first discuss signalling steps required for sensing extracellular chemoattractants such as chemokines and lipid mediators and the processes (e.g. PI3-kinase pathways) leading to the translation of extracellular chemoattractant gradients to polarized cellular responses. We will then discuss signal transduction by leucocyte adhesion receptors (e.g. tyrosine kinase pathways) which are critical for adhesion to, and migration through the vessel wall. Finally, additional neutrophil signalling pathways with an indirect effect on the neutrophil recruitment process, e.g. through modulation of the inflammatory environment, will be discussed. Mechanistic understanding of these pathways provide better understanding of the inflammation process and may point to novel therapeutic strategies for controlling excessive inflammation during infection or tissue damage. PMID:25998986

  14. Review of International Experience with Renewable Energy Obligation Support Mechanisms

    SciTech Connect

    Wiser, R.

    2005-06-01

    The main policy instruments currently used in the EU Member States to achieve the targets set for electricity produced from renewable energy sources are: (1) the quota obligation system; (2) the feed-in tariff system; and (3) the tendering system. The current study aims to review the experience gained with the quota obligation system. The report provides an overview of the regions where obligation systems have been implemented and contains a detailed evaluation of the performance of the obligation systems in the USA, the UK and in Sweden. The obligation systems in these countries have been evaluated based on the following criteria: Effectiveness; Market efficiency; Certainty for the renewable energy industry; Cost effectiveness; Stakeholder support for the obligation system; and Equity. The evaluation of international experiences with the obligation system gives rise to a mixed picture. Although an obligation in theory is effective and cost effective, it seems too early to conclude that the system delivers these promises in practice. On the one hand this is due to the limited period of implementation that makes it hard to distinguish between the direct effect of the system and some teething problems that will be solved in due time. On the other hand, the conclusion can be drawn that the obligation is a complex system, which will only function well if designed carefully. It does seem worthwhile, however, to continue monitoring the experiences with the obligation system abroad, because this will further reveal whether the system is indeed effective and cost effective in practice. In the longer term, e.g. beyond 2010, the introduction of an obligation system in the Netherlands could be considered. Finally, as the design of support schemes is being improved, it appears that the basic concepts of both the obligation system and the feed in system have been refined in such a way that the two systems are gradually converging. An important difference between the two systems however remains, namely that an obligation system relies more on market forces whereas the feed-in system is based on a greater involvement of the government.

  15. The role of TREM-2 in internalization and intracellular survival of Brucella abortus in murine macrophages.

    PubMed

    Wei, Pan; Lu, Qiang; Cui, Guimei; Guan, Zhenhong; Yang, Li; Sun, Changjiang; Sun, Wanchun; Peng, Qisheng

    2015-02-15

    Triggering receptor expressed on myeloid cells-2 (TREM-2) is a cell surface receptor primarily expressed on macrophages and dendritic cells. TREM-2 functions as a phagocytic receptor for bacteria as well as an inhibitor of Toll like receptors (TLR) induced inflammatory cytokines. However, the role of TREM-2 in Brucella intracellular growth remains unknown. To investigate whether TREM-2 is involved in Brucella intracellular survival, we chose bone marrow derived macrophages (BMDMs), in which TREM-2 is stably expressed, as cell model. Colony formation Units (CFUs) assay suggests that TREM-2 is involved in the internalization of Brucella abortus (B. abortus) by macrophages, while silencing of TREM-2 decreases intracellular survival of B. abortus. To further study the underlying mechanisms of TREM-2-mediated bacterial intracellular survival, we examined the activation of B. abortus-infected macrophages through determining the kinetics of activation of the three MAPKs, including ERK, JNK and p38, and measuring TNF? production in response to lipopolysaccharide (LPS) of Brucella (BrLPS) or B. abortus stimulation. Our data show that TREM-2 deficiency promotes activation of Brucella-infected macrophages. Moreover, our data also demonstrate that macrophage activation promotes killing of Brucella by enhancing nitric oxygen (NO), but not reactive oxygen species (ROS) production, macrophage apoptosis or cellular death. Taken together, these findings provide a novel interpretation of Brucella intracellular growth through inhibition of NO production produced by TREM-2-mediated activated macrophages. PMID:25563793

  16. Cell Extract-Containing Medium for Culture of Intracellular Fastidious Bacteria

    PubMed Central

    Singh, Sudhir; Kowalczewska, Malgorzata; Edouard, Sophie; Eldin, Carole; Perreal, Céline; Weber, Pascal; Azza, Said

    2013-01-01

    The culture of fastidious microorganisms is a critical step in infectious disease studies. As a proof-of-concept experiment, we evaluated an empirical medium containing eukaryotic cell extracts for its ability to support the growth of Coxiella burnetii. Here, we demonstrate the exponential growth of several bacterial strains, including the C. burnetii Nine Mile phase I and phase II strains, and C. burnetii isolates from humans and animals. Low-oxygen-tension conditions and the presence of small hydrophilic molecules and short peptides were critical for facilitating growth. Moreover, bacterial antigenicity was conserved, revealing the potential for this culture medium to be used in diagnostic tests and in the elaboration of vaccines against C. burnetii. We were also able to grow the majority of previously tested intracellular and fastidious bacterial species, including Tropheryma whipplei, Mycobacterium bovis, Leptospira spp., Borrelia spp., and most putative bioterrorism agents. However, we were unable to culture Rickettsia africae and Legionella spp. in this medium. The versatility of this medium should encourage its use as a replacement for the cell-based culture systems currently used for growing several facultative and putative intracellular bacterial species. PMID:23740722

  17. Identification and antimicrobial susceptibility of obligate anaerobic bacteria from clinical samples of animal origin.

    PubMed

    Mayorga, Melissa; Rodríguez-Cavallini, Evelyn; López-Ureña, Diana; Barquero-Calvo, Elías; Quesada-Gómez, Carlos

    2015-12-01

    The etiology of veterinary infectious diseases has been the focus of considerable research, yet relatively little is known about the causative agents of anaerobic infections. Susceptibility studies have documented the emergence of antimicrobial resistance and indicate distinct differences in resistance patterns related to veterinary hospitals, geographic regions, and antibiotic-prescribing regimens. The aim of the present study was to identify the obligate anaerobic bacteria from veterinary clinical samples and to determinate the in vitro susceptibility to eight antimicrobials and their resistance-associated genes. 81 clinical specimens obtained from food-producing animals, pets and wild animals were examined to determine the relative prevalence of obligate anaerobic bacteria, and the species represented. Bacteroides spp, Prevotella spp and Clostridium spp represented approximately 80% of all anaerobic isolates. Resistance to metronidazole, clindamycin, tetracycline and fluoroquinolones was found in strains isolated from food-producing animals. Ciprofloxacin, enrofloxacin and cephalotin showed the highest resistance in all isolates. In 17%, 4% and 14% of tetracycline-resistant isolates, the resistance genes tetL, tetM and tetW were respectively amplified by PCR whereas in 4% of clindamycin-resistant strains the ermG gene was detected. 26% of the isolates were positive for cepA, while only 6% harbored the cfxA (resistance-conferring genes to beta-lactams). In this study, the obligate anaerobic bacteria from Costa Rica showed a high degree of resistance to most antimicrobials tested. Nevertheless, in the majority of cases this resistance was not related to the resistance acquired genes usually described in anaerobes. It is important to address and regulate the use of antimicrobials in the agricultural industry and the empirical therapy in anaerobic bacterial infections in veterinary medicine, especially since antibiotics and resistant bacteria can persist in the environment. PMID:26385434

  18. Stochastic resonance in an intracellular genetic perceptron

    NASA Astrophysics Data System (ADS)

    Bates, Russell; Blyuss, Oleg; Zaikin, Alexey

    2014-03-01

    Intracellular genetic networks are more intelligent than was first assumed due to their ability to learn. One of the manifestations of this intelligence is the ability to learn associations of two stimuli within gene-regulating circuitry: Hebbian-type learning within the cellular life. However, gene expression is an intrinsically noisy process; hence, we investigate the effect of intrinsic and extrinsic noise on this kind of intracellular intelligence. We report a stochastic resonance in an intracellular associative genetic perceptron, a noise-induced phenomenon, which manifests itself in noise-induced increase of response in efficiency after the learning event under the conditions of optimal stochasticity.

  19. Cellular/Mollecular Intracellular Patch Electrochemistry: Regulation of Cytosolic

    E-print Network

    Sulzer, David

    Cellular/Mollecular Intracellular Patch Electrochemistry: Regulation of Cytosolic Catecholamines intracellular patch electrochemistry (IPE), a tech- nique that for the first time provides direct measurements

  20. Pregnancy Complications: Bacterial Vaginosis

    MedlinePLUS

    ... Global Map Premature birth report card Careers Archives Pregnancy Before or between pregnancies Nutrition, weight & fitness Prenatal ... complications > Bacterial vaginosis and pregnancy Bacterial vaginosis and pregnancy Now playing: E-mail to a friend Please ...

  1. Intracellular FRET-based probes: a review

    NASA Astrophysics Data System (ADS)

    Rowland, Clare E.; Brown, Carl W., III; Medintz, Igor L.; Delehanty, James B.

    2015-12-01

    Probes that exploit Förster resonance energy transfer (FRET) in their feedback mechanism are touted for their sensitivity, robustness, and low background, and thanks to the exceptional distance dependence of the energy transfer process, they provide a means of probing lengthscales well below the resolution of light. These attributes make FRET-based probes superbly suited to an intracellular environment, and recent developments in biofunctionalization and expansion of imaging capabilities have put them at the forefront of intracellular studies. Here, we present an overview of the engineering and execution of a variety of recent intracellular FRET probes, highlighting the diversity of this class of materials and the breadth of application they have found in the intracellular environment.

  2. Polymer physics of intracellular phase transitions

    NASA Astrophysics Data System (ADS)

    Brangwynne, Clifford P.; Tompa, Peter; Pappu, Rohit V.

    2015-11-01

    Intracellular organelles are either membrane-bound vesicles or membrane-less compartments that are made up of proteins and RNA. These organelles play key biological roles, by compartmentalizing the cell to enable spatiotemporal control of biological reactions. Recent studies suggest that membrane-less intracellular compartments are multicomponent viscous liquid droplets that form via phase separation. Proteins that have an intrinsic tendency for being conformationally heterogeneous seem to be the main drivers of liquid-liquid phase separation in the cell. These findings highlight the relevance of classical concepts from the physics of polymeric phase transitions for understanding the assembly of intracellular membrane-less compartments. However, applying these concepts is challenging, given the heteropolymeric nature of protein sequences, the complex intracellular environment, and non-equilibrium features intrinsic to cells. This provides new opportunities for adapting established theories and for the emergence of new physics.

  3. Introduction Many cnidarians symbiotic with the intracellular

    E-print Network

    , peroxynitrite, oxidative stress, Symbiodinium, zooxanthellae, coral bleaching, Cnidaria, di2804 Introduction Many cnidarians symbiotic with the intracellular dinoflagellate Symbiodinium sp., such as the ecologically important reef-building corals, lose their algal partners in response to a variety of stresses

  4. A Macrophage Subversion Factor Is Shared by Intracellular and Extracellular Pathogens

    PubMed Central

    Laubier, Aurélie; Bleves, Sophie; Blanc-Potard, Anne-Béatrice

    2015-01-01

    Pathogenic bacteria have developed strategies to adapt to host environment and resist host immune response. Several intracellular bacterial pathogens, including Salmonella enterica and Mycobacterium tuberculosis, share the horizontally-acquired MgtC virulence factor that is important for multiplication inside macrophages. MgtC is also found in pathogenic Pseudomonas species. Here we investigate for the first time the role of MgtC in the virulence of an extracellular pathogen, Pseudomonas aeruginosa. A P. aeruginosa mgtC mutant is attenuated in the systemic infection model of zebrafish embryos, and strikingly, the attenuated phenotype is dependent on the presence of macrophages. In ex vivo experiments, the P. aeruginosa mgtC mutant is more sensitive to macrophage killing than the wild-type strain. However, wild-type and mutant strains behave similarly toward macrophage killing when macrophages are treated with an inhibitor of the vacuolar proton ATPase. Importantly, P. aeruginosa mgtC gene expression is strongly induced within macrophages and phagosome acidification contributes to an optimal expression of the gene. Thus, our results support the implication of a macrophage intracellular stage during P. aeruginosa acute infection and suggest that Pseudomonas MgtC requires phagosome acidification to play its intracellular role. Moreover, we demonstrate that P. aeruginosa MgtC is required for optimal growth in Mg2+ deprived medium, a property shared by MgtC factors from intracellular pathogens and, under Mg2+ limitation, P. aeruginosa MgtC prevents biofilm formation. We propose that MgtC shares a similar function in intracellular and extracellular pathogens, which contributes to macrophage resistance and fine-tune adaptation to host immune response in relation to the different bacterial lifestyles. In addition, the phenotypes observed with the mgtC mutant in infection models can be mimicked in wild-type P. aeruginosa strain by producing a MgtC antagonistic peptide, thus highlighting MgtC as a promising new target for anti-virulence strategies. PMID:26080006

  5. Intracellular bacteria and adverse pregnancy outcomes.

    PubMed

    Baud, D; Greub, G

    2011-09-01

    This review considers the role of intracellular bacteria in adverse pregnancy outcomes, such as miscarriage, stillbirths, and preterm labour. The cause of miscarriage, stillbirth and preterm labour often remains unexplained. Intracellular bacteria that grow either poorly or not at all on media used routinely to detect human pathogens could be the aetiological agents of these obstetric conditions. For example, Listeria monocytogenes and Coxiella burnetti are intracellular bacteria that have a predilection for the fetomaternal unit and may induce fatal disease in the mother and/or fetus. Both are important foodborne or zoonotic pathogens in pregnancy. Preventive measures, diagnostic tools and treatment will be reviewed. Moreover, we will also address the importance in adverse pregnancy outcomes of other intracellular bacteria, including Brucella abortus and various members of the order Chlamydiales. Indeed, there is growing evidence that Chlamydia trachomatis, Chlamydia abortus and Chlamydia pneumoniae infections may also result in adverse pregnancy outcomes in humans and/or animals. Moreover, newly discovered Chlamydia-like organisms have recently emerged as new pathogens of both animals and humans. For example, Waddlia chondrophila, a Chlamydia-related bacterium isolated from aborted bovine fetuses, has also been implicated in human miscarriages. Future research should help us to better understand the pathophysiology of adverse pregnancy outcomes caused by intracellular bacteria and to determine the precise mode of transmission of newly identified bacteria, such as Waddlia and Parachlamydia. These emerging pathogens may represent the tip of the iceberg of a large number of as yet unknown intracellular pathogenic agents. PMID:21884294

  6. 31 CFR 223.18 - Performance of agency obligations.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ...Relating to Money and Finance (Continued) FISCAL SERVICE, DEPARTMENT OF THE TREASURY FINANCIAL MANAGEMENT SERVICE SURETY COMPANIES DOING BUSINESS WITH THE UNITED STATES § 223.18 Performance of agency obligations. (a)...

  7. 31 CFR 223.18 - Performance of agency obligations.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ...Relating to Money and Finance (Continued) FISCAL SERVICE, DEPARTMENT OF THE TREASURY FINANCIAL MANAGEMENT SERVICE SURETY COMPANIES DOING BUSINESS WITH THE UNITED STATES § 223.18 Performance of agency obligations. (a)...

  8. 31 CFR 223.18 - Performance of agency obligations.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ...Relating to Money and Finance (Continued) FISCAL SERVICE, DEPARTMENT OF THE TREASURY FINANCIAL MANAGEMENT SERVICE SURETY COMPANIES DOING BUSINESS WITH THE UNITED STATES § 223.18 Performance of agency obligations. (a)...

  9. 31 CFR 223.18 - Performance of agency obligations.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ...Relating to Money and Finance (Continued) FISCAL SERVICE, DEPARTMENT OF THE TREASURY FINANCIAL MANAGEMENT SERVICE SURETY COMPANIES DOING BUSINESS WITH THE UNITED STATES § 223.18 Performance of agency obligations. (a)...

  10. 7 CFR 984.54 - Establishment of obligation.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ...NUTS), DEPARTMENT OF AGRICULTURE WALNUTS GROWN IN CALIFORNIA Order Regulating Handling Reserve Walnuts § 984.54 Establishment of obligation...kernelweight of certified merchantable walnuts equal to a quantity derived by the...

  11. 7 CFR 984.54 - Establishment of obligation.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ...Nuts), DEPARTMENT OF AGRICULTURE WALNUTS GROWN IN CALIFORNIA Order Regulating Handling Reserve Walnuts § 984.54 Establishment of obligation...kernelweight of certified merchantable walnuts equal to a quantity derived by the...

  12. 7 CFR 984.54 - Establishment of obligation.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ...NUTS), DEPARTMENT OF AGRICULTURE WALNUTS GROWN IN CALIFORNIA Order Regulating Handling Reserve Walnuts § 984.54 Establishment of obligation...kernelweight of certified merchantable walnuts equal to a quantity derived by the...

  13. 7 CFR 984.54 - Establishment of obligation.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ...Nuts), DEPARTMENT OF AGRICULTURE WALNUTS GROWN IN CALIFORNIA Order Regulating Handling Reserve Walnuts § 984.54 Establishment of obligation...kernelweight of certified merchantable walnuts equal to a quantity derived by the...

  14. 7 CFR 984.54 - Establishment of obligation.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ...Nuts), DEPARTMENT OF AGRICULTURE WALNUTS GROWN IN CALIFORNIA Order Regulating Handling Reserve Walnuts § 984.54 Establishment of obligation...kernelweight of certified merchantable walnuts equal to a quantity derived by the...

  15. 22 CFR 230.07 - Fiscal Agent obligations.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...false Fiscal Agent obligations. 230.07 Section 230.07 Foreign Relations AGENCY FOR INTERNATIONAL DEVELOPMENT ISRAEL LOAN GUARANTEES ISSUED UNDER THE EMERGENCY WARTIME SUPPLEMENTAL APPROPRIATIONS ACT OF 2003, PUB. L. 108-11-STANDARD...

  16. 22 CFR 230.07 - Fiscal Agent obligations.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ...false Fiscal Agent obligations. 230.07 Section 230.07 Foreign Relations AGENCY FOR INTERNATIONAL DEVELOPMENT ISRAEL LOAN GUARANTEES ISSUED UNDER THE EMERGENCY WARTIME SUPPLEMENTAL APPROPRIATIONS ACT OF 2003, PUB. L. 108-11-STANDARD...

  17. 22 CFR 230.07 - Fiscal Agent obligations.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ...false Fiscal Agent obligations. 230.07 Section 230.07 Foreign Relations AGENCY FOR INTERNATIONAL DEVELOPMENT ISRAEL LOAN GUARANTEES ISSUED UNDER THE EMERGENCY WARTIME SUPPLEMENTAL APPROPRIATIONS ACT OF 2003, PUB. L. 108-11-STANDARD...

  18. 22 CFR 230.07 - Fiscal Agent obligations.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ...false Fiscal Agent obligations. 230.07 Section 230.07 Foreign Relations AGENCY FOR INTERNATIONAL DEVELOPMENT ISRAEL LOAN GUARANTEES ISSUED UNDER THE EMERGENCY WARTIME SUPPLEMENTAL APPROPRIATIONS ACT OF 2003, PUB. L. 108-11-STANDARD...

  19. 22 CFR 230.07 - Fiscal Agent obligations.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...false Fiscal Agent obligations. 230.07 Section 230.07 Foreign Relations AGENCY FOR INTERNATIONAL DEVELOPMENT ISRAEL LOAN GUARANTEES ISSUED UNDER THE EMERGENCY WARTIME SUPPLEMENTAL APPROPRIATIONS ACT OF 2003, PUB. L. 108-11-STANDARD...

  20. 12 CFR 612.2270 - Purchase of System obligations.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ...of Conduct § 612.2270 Purchase of System obligations...Funding Corporation, may only purchase joint, consolidated, or...with a member of the selling group designated by the Federal...Banks Funding Corporation may purchase or otherwise acquire,...

  1. 38 CFR 17.608 - Deferment of obligated service.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... (a) Request for deferment. A participant receiving a degree from a school of medicine, osteopathy, dentistry, optometry, or podiatry, may request deferment of obligated service to complete an approved program of advanced clinical...

  2. 32 CFR 220.9 - Rights and obligations of beneficiaries.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ...PARTY PAYERS OF REASONABLE CHARGES FOR HEALTHCARE SERVICES § 220.9 Rights and obligations...be required. (b) Availability of healthcare services unaffected. The availability of healthcare services in any facility of the...

  3. 12 CFR 560.42 - State and local government obligations.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... obligations. (a) What limitations apply? Pursuant to HOLA section 5(c)(1)(H), a Federal savings association... detailed analysis of credit quality. In doing so, you must consider, as appropriate, the interest...

  4. 12 CFR 560.42 - State and local government obligations.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... obligations. (a) What limitations apply? Pursuant to HOLA section 5(c)(1)(H), a Federal savings association... detailed analysis of credit quality. In doing so, you must consider, as appropriate, the interest...

  5. 12 CFR 560.42 - State and local government obligations.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... obligations. (a) What limitations apply? Pursuant to HOLA section 5(c)(1)(H), a Federal savings association... detailed analysis of credit quality. In doing so, you must consider, as appropriate, the interest...

  6. 12 CFR 560.42 - State and local government obligations.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... obligations. (a) What limitations apply? Pursuant to HOLA section 5(c)(1)(H), a Federal savings association... detailed analysis of credit quality. In doing so, you must consider, as appropriate, the interest...

  7. 12 CFR 560.42 - State and local government obligations.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... obligations. (a) What limitations apply? Pursuant to HOLA section 5(c)(1)(H), a Federal savings association... detailed analysis of credit quality. In doing so, you must consider, as appropriate, the interest...

  8. 47 CFR 25.701 - Public interest obligations.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... Telecommunication FEDERAL COMMUNICATIONS COMMISSION (CONTINUED) COMMON CARRIER SERVICES SATELLITE COMMUNICATIONS Public Interest Obligations...Entities licensed to operate satellites in the 12.2 to 12.7 GHz...

  9. 47 CFR 25.701 - Public interest obligations.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... Telecommunication FEDERAL COMMUNICATIONS COMMISSION (CONTINUED) COMMON CARRIER SERVICES SATELLITE COMMUNICATIONS Public Interest Obligations...Entities licensed to operate satellites in the 12.2 to 12.7 GHz...

  10. 47 CFR 25.701 - Public interest obligations.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... Telecommunication FEDERAL COMMUNICATIONS COMMISSION (CONTINUED) COMMON CARRIER SERVICES SATELLITE COMMUNICATIONS Public Interest Obligations...Entities licensed to operate satellites in the 12.2 to 12.7 GHz...

  11. 47 CFR 25.701 - Public interest obligations.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... Telecommunication FEDERAL COMMUNICATIONS COMMISSION (CONTINUED) COMMON CARRIER SERVICES SATELLITE COMMUNICATIONS Public Interest Obligations...Entities licensed to operate satellites in the 12.2 to 12.7 GHz...

  12. 47 CFR 54.405 - Carrier obligation to offer Lifeline.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... (CONTINUED) UNIVERSAL SERVICE Universal Service Support for Low-Income Consumers § 54.405 Carrier obligation... within the 60-day time period. A carrier providing Lifeline service in a state that has...

  13. 47 CFR 54.405 - Carrier obligation to offer Lifeline.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... (CONTINUED) UNIVERSAL SERVICE Universal Service Support for Low-Income Consumers § 54.405 Carrier obligation... demonstrate continued eligibility within the 60-day time period. A carrier providing Lifeline service in...

  14. 29 CFR 1980.102 - Obligations and prohibited acts.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ...DISCRIMINATION COMPLAINTS UNDER SECTION 806 OF THE CORPORATE AND CRIMINAL FRAUD ACCOUNTABILITY ACT OF 2002, TITLE VIII OF THE SARBANES-OXLEY ACT OF 2002 Complaints, Investigations, Findings and Preliminary Orders § 1980.102 Obligations...

  15. 29 CFR 1981.102 - Obligations and prohibited acts.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ...CONTINUED) PROCEDURES FOR THE HANDLING OF DISCRIMINATION COMPLAINTS UNDER SECTION 6 OF THE PIPELINE SAFETY IMPROVEMENT ACT OF 2002 Complaints, Investigations, Findings, and Preliminary Orders § 1981.102 Obligations and prohibited acts....

  16. 29 CFR 1981.102 - Obligations and prohibited acts.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ...CONTINUED) PROCEDURES FOR THE HANDLING OF DISCRIMINATION COMPLAINTS UNDER SECTION 6 OF THE PIPELINE SAFETY IMPROVEMENT ACT OF 2002 Complaints, Investigations, Findings, and Preliminary Orders § 1981.102 Obligations and prohibited acts....

  17. 29 CFR 1980.102 - Obligations and prohibited acts.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ...LABOR (CONTINUED) PROCEDURES FOR THE HANDLING OF RETALIATION COMPLAINTS UNDER SECTION 806 OF THE SARBANES-OXLEY ACT OF 2002, AS AMENDED Complaints, Investigations, Findings and Preliminary Orders § 1980.102 Obligations and prohibited...

  18. 29 CFR 1980.102 - Obligations and prohibited acts.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ...DISCRIMINATION COMPLAINTS UNDER SECTION 806 OF THE CORPORATE AND CRIMINAL FRAUD ACCOUNTABILITY ACT OF 2002, TITLE VIII OF THE SARBANES-OXLEY ACT OF 2002 Complaints, Investigations, Findings and Preliminary Orders § 1980.102 Obligations...

  19. 32 CFR 220.9 - Rights and obligations of beneficiaries.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ...PARTY PAYERS OF REASONABLE CHARGES FOR HEALTHCARE SERVICES § 220.9 Rights and obligations...be required. (b) Availability of healthcare services unaffected. The availability of healthcare services in any facility of the...

  20. 32 CFR 220.9 - Rights and obligations of beneficiaries.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ...PARTY PAYERS OF REASONABLE CHARGES FOR HEALTHCARE SERVICES § 220.9 Rights and obligations...be required. (b) Availability of healthcare services unaffected. The availability of healthcare services in any facility of the...

  1. 32 CFR 220.9 - Rights and obligations of beneficiaries.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ...PARTY PAYERS OF REASONABLE CHARGES FOR HEALTHCARE SERVICES § 220.9 Rights and obligations...be required. (b) Availability of healthcare services unaffected. The availability of healthcare services in any facility of the...

  2. 32 CFR 220.9 - Rights and obligations of beneficiaries.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ...PARTY PAYERS OF REASONABLE CHARGES FOR HEALTHCARE SERVICES § 220.9 Rights and obligations...be required. (b) Availability of healthcare services unaffected. The availability of healthcare services in any facility of the...

  3. 34 CFR 686.43 - Obligation to repay the grant.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ...Offices of the Department of Education (Continued) OFFICE OF POSTSECONDARY EDUCATION, DEPARTMENT OF EDUCATION (CONTINUED) TEACHER EDUCATION ASSISTANCE FOR COLLEGE AND HIGHER EDUCATION (TEACH) GRANT PROGRAM Service and Repayment Obligations §...

  4. 34 CFR 686.43 - Obligation to repay the grant.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ...Offices of the Department of Education (Continued) OFFICE OF POSTSECONDARY EDUCATION, DEPARTMENT OF EDUCATION (CONTINUED) TEACHER EDUCATION ASSISTANCE FOR COLLEGE AND HIGHER EDUCATION (TEACH) GRANT PROGRAM Service and Repayment Obligations §...

  5. 34 CFR 686.43 - Obligation to repay the grant.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ...Regulations of the Offices of the Department of Education (Continued) OFFICE OF POSTSECONDARY EDUCATION, DEPARTMENT OF EDUCATION TEACHER EDUCATION ASSISTANCE FOR COLLEGE AND HIGHER EDUCATION (TEACH) GRANT PROGRAM Service and Repayment Obligations §...

  6. 34 CFR 686.43 - Obligation to repay the grant.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ...Offices of the Department of Education (Continued) OFFICE OF POSTSECONDARY EDUCATION, DEPARTMENT OF EDUCATION (CONTINUED) TEACHER EDUCATION ASSISTANCE FOR COLLEGE AND HIGHER EDUCATION (TEACH) GRANT PROGRAM Service and Repayment Obligations §...

  7. 34 CFR 686.43 - Obligation to repay the grant.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ...Offices of the Department of Education (Continued) OFFICE OF POSTSECONDARY EDUCATION, DEPARTMENT OF EDUCATION (CONTINUED) TEACHER EDUCATION ASSISTANCE FOR COLLEGE AND HIGHER EDUCATION (TEACH) GRANT PROGRAM Service and Repayment Obligations §...

  8. 23 CFR 450.332 - Annual listing of obligated projects.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...OF TRANSPORTATION PLANNING AND RESEARCH PLANNING ASSISTANCE AND STANDARDS Metropolitan Transportation Planning and Programming § 450.332 Annual listing of obligated projects. (a) In metropolitan planning areas, on an annual...

  9. 7 CFR 760.507 - Obligations of a participant.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ...PROGRAMS INDEMNITY PAYMENT PROGRAMS Tree Assistance Program § 760.507 Obligations... (a) Eligible orchardists and nursery tree growers must execute all required documents... (b) Eligible orchardist or nursery tree growers must allow representatives...

  10. 7 CFR 760.507 - Obligations of a participant.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ...PROGRAMS INDEMNITY PAYMENT PROGRAMS Tree Assistance Program § 760.507 Obligations... (a) Eligible orchardists and nursery tree growers must execute all required documents... (b) Eligible orchardist or nursery tree growers must allow representatives...

  11. 7 CFR 760.507 - Obligations of a participant.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ...PROGRAMS INDEMNITY PAYMENT PROGRAMS Tree Assistance Program § 760.507 Obligations... (a) Eligible orchardists and nursery tree growers must execute all required documents... (b) Eligible orchardist or nursery tree growers must allow representatives...

  12. 7 CFR 783.7 - Obligations of a participant.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ...Regulations of the Department of Agriculture (Continued) FARM SERVICE AGENCY, DEPARTMENT OF AGRICULTURE SPECIAL PROGRAMS TREE ASSISTANCE PROGRAM § 783.7 Obligations of a participant. (a) Eligible orchardists must execute all required...

  13. 7 CFR 760.507 - Obligations of a participant.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ...PROGRAMS INDEMNITY PAYMENT PROGRAMS Tree Assistance Program § 760.507 Obligations... (a) Eligible orchardists and nursery tree growers must execute all required documents... (b) Eligible orchardist or nursery tree growers must allow representatives...

  14. 16 CFR 436.2 - Obligation to furnish documents.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ...Obligation to furnish documents. In connection with the offer or sale of a franchise...current disclosure document, as described...proposed franchise sale. (b) For any...to the disclosure document without...

  15. 16 CFR 436.2 - Obligation to furnish documents.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ...Obligation to furnish documents. In connection with the offer or sale of a franchise...current disclosure document, as described...proposed franchise sale. (b) For any...to the disclosure document without...

  16. 16 CFR 436.2 - Obligation to furnish documents.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ...Obligation to furnish documents. In connection with the offer or sale of a franchise...current disclosure document, as described...proposed franchise sale. (b) For any...to the disclosure document without...

  17. 16 CFR 436.2 - Obligation to furnish documents.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ...Obligation to furnish documents. In connection with the offer or sale of a franchise...current disclosure document, as described...proposed franchise sale. (b) For any...to the disclosure document without...

  18. 16 CFR 436.2 - Obligation to furnish documents.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ...Obligation to furnish documents. In connection with the offer or sale of a franchise...current disclosure document, as described...proposed franchise sale. (b) For any...to the disclosure document without...

  19. Towards Regulatory Compliance: Extracting Rights and Obligation to

    E-print Network

    Breaux, Travis D.

    Towards Regulatory Compliance: Extracting Rights and Obligation to Align Requirements Regulations and Requirements Traceability in Legal Language Modeling Regulatory Semantics Case Study: HIPAA: From Regulations to Requirements Regulations govern the system "environment." Regulatory language

  20. Polymer-Ag nanocomposites with enhanced antimicrobial activity against bacterial infection.

    PubMed

    Mei, Lin; Lu, Zhentan; Zhang, Xinge; Li, Chaoxing; Jia, Yanxia

    2014-09-24

    Herein, a nontoxic nanocomposite is synthesized by reduction of silver nitrate in the presence of a cationic polymer displaying strong antimicrobial activity against bacterial infection. These nanocomposites with a large concentration of positive charge promote their adsorption to bacterial membranes through electrostatic interaction. Moreover, the synthesized nanocomposites with polyvalent and synergistic antimicrobial effects can effectively kill both Gram-positive and Gram-negative bacteria without the emergence of bacterial resistance. Morphological changes obtained by transmission electron microscope observation show that these nanocomposites can cause leakage and chaos of intracellular contents. Analysis of the antimicrobial mechanism confirms that the lethal action of nanocomposites against the bacteria started with disruption of the bacterial membrane, subsequent cellular internalization of the nanoparticles, and inhibition of intracellular enzymatic activity. This novel antimicrobial material with good cytocompatibility promotes healing of infected wounds in diabetic rats, and has a promising future in the treatment of other infectious diseases. PMID:25170799

  1. Efficient intracellular delivery and improved biocompatibility of colloidal silver nanoparticles towards intracellular SERS immuno-sensing.

    PubMed

    Bhardwaj, Vinay; Srinivasan, Supriya; McGoron, Anthony J

    2015-06-21

    High throughput intracellular delivery strategies, electroporation, passive and TATHA2 facilitated diffusion of colloidal silver nanoparticles (AgNPs) are investigated for cellular toxicity and uptake using state-of-art analytical techniques. The TATHA2 facilitated approach efficiently delivered high payload with no toxicity, pre-requisites for intracellular applications of plasmonic metal nanoparticles (PMNPs) in sensing and therapeutics. PMID:25939798

  2. The COPII complex and lysosomal VAMP7 determine intracellular Salmonella localization and growth.

    PubMed

    Santos, José Carlos; Duchateau, Magalie; Fredlund, Jennifer; Weiner, Allon; Mallet, Adeline; Schmitt, Christine; Matondo, Mariette; Hourdel, Véronique; Chamot-Rooke, Julia; Enninga, Jost

    2015-12-01

    Salmonella invades epithelial cells and survives within a membrane-bound compartment, the Salmonella-containing vacuole (SCV). We isolated and determined the host protein composition of the SCV at 30?min and 3?h of infection to identify and characterize novel regulators of intracellular bacterial localization and growth. Quantitation of the SCV protein content revealed 392 host proteins specifically enriched at SCVs, out of which 173 associated exclusively with early SCVs, 124 with maturing SCV and 95 proteins during both time-points. Vacuole interactions with endoplasmic reticulum-derived coat protein complex II vesicles modulate early steps of SCV maturation, promoting SCV rupture and bacterial hyper-replication within the host cytosol. On the other hand, SCV interactions with VAMP7-positive lysosome-like vesicles promote Salmonella-induced filament formation and bacterial growth within the late SCV. Our results reveal that the dynamic communication between the SCV and distinct host organelles affects both intracellular Salmonella localization and growth at successive steps of host cell invasion. PMID:26084942

  3. Chemical development of intracellular protein heterodimerizers.

    PubMed

    Erhart, Dominik; Zimmermann, Mirjam; Jacques, Olivier; Wittwer, Matthias B; Ernst, Beat; Constable, Edwin; Zvelebil, Marketa; Beaufils, Florent; Wymann, Matthias P

    2013-04-18

    Cell activation initiated by receptor ligands or oncogenes triggers complex and convoluted intracellular signaling. Techniques initiating signals at defined starting points and cellular locations are attractive to elucidate the output of selected pathways. Here, we present the development and validation of a protein heterodimerization system based on small molecules cross-linking fusion proteins derived from HaloTags and SNAP-tags. Chemical dimerizers of HaloTag and SNAP-tag (HaXS) show excellent selectivity and have been optimized for intracellular reactivity. HaXS force protein-protein interactions and can translocate proteins to various cellular compartments. Due to the covalent nature of the HaloTag-HaXS-SNAP-tag complex, intracellular dimerization can be easily monitored. First applications include protein targeting to cytoskeleton, to the plasma membrane, to lysosomes, the initiation of the PI3K/mTOR pathway, and multiplexed protein complex formation in combination with the rapamycin dimerization system. PMID:23601644

  4. Intracellular metabolite levels shape sulfur isotope fractionation during microbial sulfate respiration

    PubMed Central

    Wing, Boswell A.; Halevy, Itay

    2014-01-01

    We present a quantitative model for sulfur isotope fractionation accompanying bacterial and archaeal dissimilatory sulfate respiration. By incorporating independently available biochemical data, the model can reproduce a large number of recent experimental fractionation measurements with only three free parameters: (i) the sulfur isotope selectivity of sulfate uptake into the cytoplasm, (ii) the ratio of reduced to oxidized electron carriers supporting the respiration pathway, and (iii) the ratio of in vitro to in vivo levels of respiratory enzyme activity. Fractionation is influenced by all steps in the dissimilatory pathway, which means that environmental sulfate and sulfide levels control sulfur isotope fractionation through the proximate influence of intracellular metabolites. Although sulfur isotope fractionation is a phenotypic trait that appears to be strain specific, we show that it converges on near-thermodynamic behavior, even at micromolar sulfate levels, as long as intracellular sulfate reduction rates are low enough (<<1 fmol H2S?cell?1?d?1). PMID:25362045

  5. Fast, Ultrasensitive Detection of Reactive Oxygen Species Using a Carbon Nanotube Based-Electrocatalytic Intracellular Sensor.

    PubMed

    Rawson, Frankie J; Hicks, Jacqueline; Dodd, Nicholas; Abate, Wondwossen; Garrett, David J; Yip, Nga; Fejer, Gyorgy; Downard, Alison J; Baronian, Kim H R; Jackson, Simon K; Mendes, Paula M

    2015-10-28

    Herein, we report a highly sensitive electrocatalytic sensor-cell construct that can electrochemically communicate with the internal environment of immune cells (e.g., macrophages) via the selective monitoring of a particular reactive oxygen species (ROS), hydrogen peroxide. The sensor, which is based on vertically aligned single-walled carbon nanotubes functionalized with an osmium electrocatalyst, enabled the unprecedented detection of a local intracellular "pulse" of ROS on a short second time scale in response to bacterial endotoxin (lipopolysaccharide-LPS) stimulation. Our studies have shown that this initial pulse of ROS is dependent on NADPH oxidase (NOX) and toll like receptor 4 (TLR4). The results suggest that bacteria can induce a rapid intracellular pulse of ROS in macrophages that initiates the classical innate immune response of these cells to infection. PMID:26438964

  6. Fast, Ultrasensitive Detection of Reactive Oxygen Species Using a Carbon Nanotube Based-Electrocatalytic Intracellular Sensor

    PubMed Central

    2015-01-01

    Herein, we report a highly sensitive electrocatalytic sensor-cell construct that can electrochemically communicate with the internal environment of immune cells (e.g., macrophages) via the selective monitoring of a particular reactive oxygen species (ROS), hydrogen peroxide. The sensor, which is based on vertically aligned single-walled carbon nanotubes functionalized with an osmium electrocatalyst, enabled the unprecedented detection of a local intracellular “pulse” of ROS on a short second time scale in response to bacterial endotoxin (lipopolysaccharide-LPS) stimulation. Our studies have shown that this initial pulse of ROS is dependent on NADPH oxidase (NOX) and toll like receptor 4 (TLR4). The results suggest that bacteria can induce a rapid intracellular pulse of ROS in macrophages that initiates the classical innate immune response of these cells to infection. PMID:26438964

  7. Solving a Bloody Mess: B-Vitamin Independent Metabolic Convergence among Gammaproteobacterial Obligate Endosymbionts from Blood-Feeding Arthropods and the Leech Haementeria officinalis

    PubMed Central

    Manzano-Marín, Alejandro; Oceguera-Figueroa, Alejandro; Latorre, Amparo; Jiménez-García, Luis F.; Moya, Andres

    2015-01-01

    Endosymbiosis is a common phenomenon in nature, especially between bacteria and insects, whose typically unbalanced diets are usually complemented by their obligate endosymbionts. While much interest and focus has been directed toward phloem-feeders like aphids and mealybugs, blood-feeders such as the Lone star tick (Amblyomma americanum), Glossina flies, and the human body louse (Pediculus humanus corporis) depend on obligate endosymbionts which complement their B-vitamin-deficient diets, and thus are required for growth and survival. Glossiphoniid leeches have also been found to harbor distinct endosymbionts housed in specialized organs. Here, we present the genome of the bacterial endosymbiont from Haementeria officinalis, first of a glossiphoniid leech. This as-yet-unnamed endosymbiont belongs to the Gammaproteobacteria, has a pleomorphic shape and is restricted to bacteriocytes. For this bacterial endosymbiont, we propose the name Candidatus Providencia siddallii. This symbiont possesses a highly reduced genome with high A+T content and a reduced set of metabolic capabilities, all of which are common characteristics of ancient obligate endosymbionts of arthropods. Its genome has retained many pathways related to the biosynthesis of B-vitamins, pointing toward a role in supplementing the blood-restricted diet of its host. Through comparative genomics against the endosymbionts of A. americanum, Glossina flies, and P. humanus corporis, we were able to detect a high degree of metabolic convergence among these four very distantly related endosymbiotic bacteria. PMID:26454017

  8. Solving a Bloody Mess: B-Vitamin Independent Metabolic Convergence among Gammaproteobacterial Obligate Endosymbionts from Blood-Feeding Arthropods and the Leech Haementeria officinalis.

    PubMed

    Manzano-Marín, Alejandro; Oceguera-Figueroa, Alejandro; Latorre, Amparo; Jiménez-García, Luis F; Moya, Andres

    2015-10-01

    Endosymbiosis is a common phenomenon in nature, especially between bacteria and insects, whose typically unbalanced diets are usually complemented by their obligate endosymbionts. While much interest and focus has been directed toward phloem-feeders like aphids and mealybugs, blood-feeders such as the Lone star tick (Amblyomma americanum), Glossina flies, and the human body louse (Pediculus humanus corporis) depend on obligate endosymbionts which complement their B-vitamin-deficient diets, and thus are required for growth and survival. Glossiphoniid leeches have also been found to harbor distinct endosymbionts housed in specialized organs. Here, we present the genome of the bacterial endosymbiont from Haementeria officinalis, first of a glossiphoniid leech. This as-yet-unnamed endosymbiont belongs to the Gammaproteobacteria, has a pleomorphic shape and is restricted to bacteriocytes. For this bacterial endosymbiont, we propose the name Candidatus Providencia siddallii. This symbiont possesses a highly reduced genome with high A+T content and a reduced set of metabolic capabilities, all of which are common characteristics of ancient obligate endosymbionts of arthropods. Its genome has retained many pathways related to the biosynthesis of B-vitamins, pointing toward a role in supplementing the blood-restricted diet of its host. Through comparative genomics against the endosymbionts of A. americanum, Glossina flies, and P. humanus corporis, we were able to detect a high degree of metabolic convergence among these four very distantly related endosymbiotic bacteria. PMID:26454017

  9. Asphalt Showing Bacterial Degeneration 

    E-print Network

    Unknown

    2011-08-17

    , has been expanded through a new project, Development of a Synergistic, Comprehensive Statewide Lone Star Healthy Streams Program. The LSHS Program initially developed and tested educational opportunities for cattlemen focusing on bacterial... contami- nation of watersheds by grazing animals and how that bacterial contamination can be reduced. The program also encouraged adoption of best management practices designed to reduce bacterial loading to Texas streams and waterways...

  10. Facial bacterial infections: folliculitis.

    PubMed

    Laureano, Ana Cristina; Schwartz, Robert A; Cohen, Philip J

    2014-01-01

    Facial bacterial infections are most commonly caused by infections of the hair follicles. Wherever pilosebaceous units are found folliculitis can occur, with the most frequent bacterial culprit being Staphylococcus aureus. We review different origins of facial folliculitis, distinguishing bacterial forms from other infectious and non-infectious mimickers. We distinguish folliculitis from pseudofolliculitis and perifolliculitis. Clinical features, etiology, pathology, and management options are also discussed. PMID:25441463

  11. Coxiella burnetii Effector Proteins That Localize to the Parasitophorous Vacuole Membrane Promote Intracellular Replication

    PubMed Central

    Larson, Charles L.; Beare, Paul A.; Voth, Daniel E.; Howe, Dale; Cockrell, Diane C.; Bastidas, Robert J.; Valdivia, Raphael H.

    2014-01-01

    The intracellular bacterial pathogen Coxiella burnetii directs biogenesis of a parasitophorous vacuole (PV) that acquires host endolysosomal components. Formation of a PV that supports C. burnetii replication requires a Dot/Icm type 4B secretion system (T4BSS) that delivers bacterial effector proteins into the host cell cytosol. Thus, a subset of T4BSS effectors are presumed to direct PV biogenesis. Recently, the PV-localized effector protein CvpA was found to promote C. burnetii intracellular growth and PV expansion. We predict additional C. burnetii effectors localize to the PV membrane and regulate eukaryotic vesicle trafficking events that promote pathogen growth. To identify these vacuolar effector proteins, a list of predicted C. burnetii T4BSS substrates was compiled using bioinformatic criteria, such as the presence of eukaryote-like coiled-coil domains. Adenylate cyclase translocation assays revealed 13 proteins were secreted in a Dot/Icm-dependent fashion by C. burnetii during infection of human THP-1 macrophages. Four of the Dot/Icm substrates, termed Coxiella vacuolar protein B (CvpB), CvpC, CvpD, and CvpE, labeled the PV membrane and LAMP1-positive vesicles when ectopically expressed as fluorescently tagged fusion proteins. C. burnetii ?cvpB, ?cvpC, ?cvpD, and ?cvpE mutants exhibited significant defects in intracellular replication and PV formation. Genetic complementation of the ?cvpD and ?cvpE mutants rescued intracellular growth and PV generation, whereas the growth of C. burnetii ?cvpB and ?cvpC was rescued upon cohabitation with wild-type bacteria in a common PV. Collectively, these data indicate C. burnetii encodes multiple effector proteins that target the PV membrane and benefit pathogen replication in human macrophages. PMID:25422265

  12. Coxiella burnetii effector proteins that localize to the parasitophorous vacuole membrane promote intracellular replication.

    PubMed

    Larson, Charles L; Beare, Paul A; Voth, Daniel E; Howe, Dale; Cockrell, Diane C; Bastidas, Robert J; Valdivia, Raphael H; Heinzen, Robert A

    2015-02-01

    The intracellular bacterial pathogen Coxiella burnetii directs biogenesis of a parasitophorous vacuole (PV) that acquires host endolysosomal components. Formation of a PV that supports C. burnetii replication requires a Dot/Icm type 4B secretion system (T4BSS) that delivers bacterial effector proteins into the host cell cytosol. Thus, a subset of T4BSS effectors are presumed to direct PV biogenesis. Recently, the PV-localized effector protein CvpA was found to promote C. burnetii intracellular growth and PV expansion. We predict additional C. burnetii effectors localize to the PV membrane and regulate eukaryotic vesicle trafficking events that promote pathogen growth. To identify these vacuolar effector proteins, a list of predicted C. burnetii T4BSS substrates was compiled using bioinformatic criteria, such as the presence of eukaryote-like coiled-coil domains. Adenylate cyclase translocation assays revealed 13 proteins were secreted in a Dot/Icm-dependent fashion by C. burnetii during infection of human THP-1 macrophages. Four of the Dot/Icm substrates, termed Coxiella vacuolar protein B (CvpB), CvpC, CvpD, and CvpE, labeled the PV membrane and LAMP1-positive vesicles when ectopically expressed as fluorescently tagged fusion proteins. C. burnetii ?cvpB, ?cvpC, ?cvpD, and ?cvpE mutants exhibited significant defects in intracellular replication and PV formation. Genetic complementation of the ?cvpD and ?cvpE mutants rescued intracellular growth and PV generation, whereas the growth of C. burnetii ?cvpB and ?cvpC was rescued upon cohabitation with wild-type bacteria in a common PV. Collectively, these data indicate C. burnetii encodes multiple effector proteins that target the PV membrane and benefit pathogen replication in human macrophages. PMID:25422265

  13. The obligate mutualist Wigglesworthia glossinidia influences reproduction, digestion, and immunity processes of its host, the tsetse fly.

    PubMed

    Pais, Roshan; Lohs, Claudia; Wu, Yineng; Wang, Jingwen; Aksoy, Serap

    2008-10-01

    Tsetse flies (Diptera: Glossinidae) are vectors for trypanosome parasites, the agents of the deadly sleeping sickness disease in Africa. Tsetse also harbor two maternally transmitted enteric mutualist endosymbionts: the primary intracellular obligate Wigglesworthia glossinidia and the secondary commensal Sodalis glossinidius. Both endosymbionts are transmitted to the intrauterine progeny through the milk gland secretions of the viviparous female. We administered various antibiotics either continuously by per os supplementation of the host blood meal diet or discretely by hemocoelic injections into fertile females in an effort to selectively eliminate the symbionts to study their individual functions. A symbiont-specific PCR amplification assay and fluorescence in situ hybridization analysis were used to evaluate symbiont infection outcomes. Tetracycline and rifampin treatments eliminated all tsetse symbionts but reduced the fecundity of the treated females. Ampicillin treatments did not affect the intracellular Wigglesworthia localized in the bacteriome organ and retained female fecundity. The resulting progeny of ampicillin-treated females, however, lacked Wigglesworthia but still harbored the commensal Sodalis. Our results confirm the presence of two physiologically distinct Wigglesworthia populations: the bacteriome-localized Wigglesworthia involved with nutritional symbiosis and free-living Wigglesworthia in the milk gland organ responsible for maternal transmission to the progeny. We evaluated the reproductive fitness, longevity, digestion, and vectorial competence of flies that were devoid of Wigglesworthia. The absence of Wigglesworthia completely abolished the fertility of females but not that of males. Both the male and female Wigglesworthia-free adult progeny displayed longevity costs and were significantly compromised in their blood meal digestion ability. Finally, while the vectorial competence of the young newly hatched adults without Wigglesworthia was comparable to that of their wild-type counterparts, older flies displayed higher susceptibility to trypanosome infections, indicating a role for the mutualistic symbiosis in host immunobiology. The ability to rear adult tsetse that lack the obligate Wigglesworthia endosymbionts will now enable functional investigations into this ancient symbiosis. PMID:18689507

  14. Diversity of extracellular proteins during the transition from the 'proto-apicomplexan' alveolates to the apicomplexan obligate parasites.

    PubMed

    Templeton, Thomas J; Pain, Arnab

    2016-01-01

    The recent completion of high-coverage draft genome sequences for several alveolate protozoans - namely, the chromerids, Chromera velia and Vitrella brassicaformis; the perkinsid Perkinsus marinus; the apicomplexan, Gregarina niphandrodes, as well as high coverage transcriptome sequence information for several colpodellids, allows for new genome-scale comparisons across a rich landscape of apicomplexans and other alveolates. Genome annotations can now be used to help interpret fine ultrastructure and cell biology, and guide new studies to describe a variety of alveolate life strategies, such as symbiosis or free living, predation, and obligate intracellular parasitism, as well to provide foundations to dissect the evolutionary transitions between these niches. This review focuses on the attempt to identify extracellular proteins which might mediate the physical interface of cell-cell interactions within the above life strategies, aided by annotation of the repertoires of predicted surface and secreted proteins encoded within alveolate genomes. In particular, we discuss what descriptions of the predicted extracellular proteomes reveal regarding a hypothetical last common ancestor of a pre-apicomplexan alveolate - guided by ultrastructure, life strategies and phylogenetic relationships - in an attempt to understand the evolution of obligate parasitism in apicomplexans. PMID:26585326

  15. Ion channels enable electrical communication in bacterial communities.

    PubMed

    Prindle, Arthur; Liu, Jintao; Asally, Munehiro; Ly, San; Garcia-Ojalvo, Jordi; Süel, Gürol M

    2015-11-01

    The study of bacterial ion channels has provided fundamental insights into the structural basis of neuronal signalling; however, the native role of ion channels in bacteria has remained elusive. Here we show that ion channels conduct long-range electrical signals within bacterial biofilm communities through spatially propagating waves of potassium. These waves result from a positive feedback loop, in which a metabolic trigger induces release of intracellular potassium, which in turn depolarizes neighbouring cells. Propagating through the biofilm, this wave of depolarization coordinates metabolic states among cells in the interior and periphery of the biofilm. Deletion of the potassium channel abolishes this response. As predicted by a mathematical model, we further show that spatial propagation can be hindered by specific genetic perturbations to potassium channel gating. Together, these results demonstrate a function for ion channels in bacterial biofilms, and provide a prokaryotic paradigm for active, long-range electrical signalling in cellular communities. PMID:26503040

  16. Neutrophil cell surface receptors and their intracellular signal transduction pathways?

    PubMed Central

    Futosi, Krisztina; Fodor, Szabina; Mócsai, Attila

    2013-01-01

    Neutrophils play a critical role in the host defense against bacterial and fungal infections, but their inappropriate activation also contributes to tissue damage during autoimmune and inflammatory diseases. Neutrophils express a large number of cell surface receptors for the recognition of pathogen invasion and the inflammatory environment. Those include G-protein-coupled chemokine and chemoattractant receptors, Fc-receptors, adhesion receptors such as selectins/selectin ligands and integrins, various cytokine receptors, as well as innate immune receptors such as Toll-like receptors and C-type lectins. The various cell surface receptors trigger very diverse signal transduction pathways including activation of heterotrimeric and monomeric G-proteins, receptor-induced and store-operated Ca2 + signals, protein and lipid kinases, adapter proteins and cytoskeletal rearrangement. Here we provide an overview of the receptors involved in neutrophil activation and the intracellular signal transduction processes they trigger. This knowledge is crucial for understanding how neutrophils participate in antimicrobial host defense and inflammatory tissue damage and may also point to possible future targets of the pharmacological therapy of neutrophil-mediated autoimmune or inflammatory diseases. PMID:23994464

  17. Intracellular trafficking of Mycobacterium avium ss. paratuberculosis in macrophages.

    PubMed

    Cheville, N F; Hostetter, J; Thomsen, B V; Simutis, F; Vanloubbeeck, Y; Steadham, E

    2001-06-01

    The granulomatous enteric lesions of cattle with Johne's disease are composed of infected macrophages, and grow by accumulation, re-infection, and expansion of macrophage populations in the intestinal wall. We have examined the growth of bacteria in macrophages to define characteristics of intracellular trafficking for exocytosis, replication, and antigen presentation. Using immunocytochemical markers for light, confocal and electron microscopy, we have examined potential pathway tropisms using data for bacterial attachment, phagosomal acidification, phagolysosomal degradation and apoptosis. Our hypotheses are that pathogenic/wild-type strains block phagosomal acidification so that the phagosome fails to obtain markers of the late phagosome and phagolysosome, and this leads to the replication pathway within bacteriophorous vacuoles. Non-pathogenic strains appear to be processed to exocytosis, and avirulent mutant strains may be degraded and have preference of antigen processing pathways that involve transport vesicles bearing MHC II antigens. Pathogenicity in a nude mouse model of intestinal infection reveals lesion development and confirms pathway preferences of virulent strains for bacteriophorous vacuole formation. PMID:11449907

  18. Identification of Bacterial Populations in Drinking Water Using 16S rRNA-Based Sequence Analyses

    EPA Science Inventory

    Intracellular RNA is rapidly degraded in stressed cells and is more unstable outside of the cell than DNA. As a result, RNA-based methods have been suggested to study the active microbial fraction in environmental matrices. The aim of this study was to identify bacterial populati...

  19. Metablic Coevolution in the bacterial symbiosis of whiteflies and related plant sap-feeding insects

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In animals dependent on intracellular bacteria with very small genomes, the host cell is adapted to support the function of its bacterial symbionts, but the molecular basis of these adaptations is poorly understood. We investigated the metabolic coevolution between the whitefly Bemisia tabaci and th...

  20. Physical Features of Intracellular Proteins that Moonlight on the Cell Surface

    PubMed Central

    Amblee, Vaishak; Jeffery, Constance J.

    2015-01-01

    Moonlighting proteins comprise a subset of multifunctional proteins that perform two or more biochemical functions that are not due to gene fusions, multiple splice variants, proteolytic fragments, or promiscuous enzyme activities. The project described herein focuses on a sub-set of moonlighting proteins that have a canonical biochemical function inside the cell and perform a second biochemical function on the cell surface in at least one species. The goal of this project is to consider the biophysical features of these moonlighting proteins to determine whether they have shared characteristics or defining features that might suggest why these particular proteins were adopted for a second function on the cell surface, or if these proteins resemble typical intracellular proteins. The latter might suggest that many other normally intracellular proteins found on the cell surface might also be moonlighting in this fashion. We have identified 30 types of proteins that have different functions inside the cell and on the cell surface. Some of these proteins are found to moonlight on the surface of multiple species, sometimes with different extracellular functions in different species, so there are a total of 98 proteins in the study set. Although a variety of intracellular proteins (enzymes, chaperones, etc.) are observed to be re-used on the cell surface, for the most part, these proteins were found to have physical characteristics typical of intracellular proteins. Many other intracellular proteins have also been found on the surface of bacterial pathogens and other organisms in proteomics experiments. It is quite possible that many of those proteins also have a moonlighting function on the cell surface. The increasing number and variety of known moonlighting proteins suggest that there may be more moonlighting proteins than previously thought, and moonlighting might be a common feature of many more proteins. PMID:26110848

  1. Analysis of the Proteome of Intracellular Shigella flexneri Reveals Pathways Important for Intracellular Growth

    PubMed Central

    Pieper, Rembert; Fisher, C. R.; Suh, Moo-Jin; Huang, S.-T.; Parmar, P.

    2013-01-01

    Global proteomic analysis was performed with Shigella flexneri strain 2457T in association with three distinct growth environments: S. flexneri growing in broth (in vitro), S. flexneri growing within epithelial cell cytoplasm (intracellular), and S. flexneri that were cultured with, but did not invade, Henle cells (extracellular). Compared to in vitro and extracellular bacteria, intracellular bacteria had increased levels of proteins required for invasion and cell-to-cell spread, including Ipa, Mxi, and Ics proteins. Changes in metabolic pathways in response to the intracellular environment also were evident. There was an increase in glycogen biosynthesis enzymes, altered expression of sugar transporters, and a reduced amount of the carbon storage regulator CsrA. Mixed acid fermentation enzymes were highly expressed intracellularly, while tricarboxylic acid (TCA) cycle oxidoreductive enzymes and most electron transport chain proteins, except CydAB, were markedly decreased. This suggested that fermentation and the CydAB system primarily sustain energy generation intracellularly. Elevated levels of PntAB, which is responsible for NADPH regeneration, suggested a shortage of reducing factors for ATP synthesis. These metabolic changes likely reflect changes in available carbon sources, oxygen levels, and iron availability. Intracellular bacteria showed strong evidence of iron starvation. Iron acquisition systems (Iut, Sit, FhuA, and Feo) and the iron starvation, stress-associated Fe-S cluster assembly (Suf) protein were markedly increased in abundance. Mutational analysis confirmed that the mixed-acid fermentation pathway was required for wild-type intracellular growth and spread of S. flexneri. Thus, iron stress and changes in carbon metabolism may be key factors in the S. flexneri transition from the extra- to the intracellular milieu. PMID:24101689

  2. The intracellular citrus huanglongbing bacterium, 'Candidatus Liberibacter asiaticus' encodes two novel autotransporters.

    PubMed

    Hao, Guixia; Boyle, Michael; Zhou, Lijuan; Duan, Yongping

    2013-01-01

    Proteins secreted by the type V secretion system (T5SS), known as autotransporters, are large extracellular virulence proteins localized to the bacterial poles. In this study, we characterized two novel autotransporter proteins of 'Candidatus Liberibacter asiaticus' (Las), and redesignated them as LasAI and LasAII in lieu of the previous names HyvI and HyvII. As a phloem-limited, intracellular bacterial pathogen, Las has a significantly reduced genome and causes huanglongbing (HLB), a devastating disease of citrus worldwide. Bioinformatic analyses revealed that LasAI and LasAII share the structural features of an autotransporter family containing large repeats of a passenger domain and a unique C-terminal translocator domain. When fused to the GFP gene and expressed in E. coli, the LasAI C-terminus and the full length LasAII were localized to the bacterial poles, similar to other members of autotransporter family. Despite the absence of a typical signal peptide, LasAI was found to localize at the cell surface by immuno-dot blot using a monoclonal antibody against the partial LasAI protein. Its surface localization was also confirmed by the removal of the LasAI antigen using a proteinase K treatment of the intact bacterial cells. When co-inoculated with a P19 gene silencing suppressor and transiently expressed in tobacco leaves, the GFP-LasAI translocator targeted to the mitochondria. This is the first report that Las encodes novel autotransporters that target to mitochondria when expressed in the plants. These findings may lead to a better understanding of the pathogenesis of this intracellular bacterium. PMID:23874813

  3. 76 FR 16707 - Rule 17Ad-17; Transfer Agents', Brokers', and Dealers' Obligation To Search for Lost...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-03-25

    ...Brokers', and Dealers' Obligation To Search for Lost Securityholders; Paying Agents' Obligation To Search for Missing Securityholders AGENCY: Securities...Transfer Agents' Obligation to Search for Lost Securityholders'' to:...

  4. 31 CFR 225.3 - Pledge of Government obligations in lieu of a bond with surety or sureties.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ...2011-07-01 false Pledge of Government obligations in lieu of a bond with... ACCEPTANCE OF BONDS SECURED BY GOVERNMENT OBLIGATIONS IN LIEU OF BONDS WITH SURETIES § 225.3 Pledge of Government obligations in lieu of a bond...

  5. 31 CFR 225.3 - Pledge of Government obligations in lieu of a bond with surety or sureties.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ...2010-07-01 false Pledge of Government obligations in lieu of a bond with... ACCEPTANCE OF BONDS SECURED BY GOVERNMENT OBLIGATIONS IN LIEU OF BONDS WITH SURETIES § 225.3 Pledge of Government obligations in lieu of a bond...

  6. 26 CFR 1.165-12 - Denial of deduction for losses on registration-required obligations not in registered form.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...or obligation (including conversion privilege) was issued after...customer, custodial or nominee relationship and such institution agrees...such paragraph. (4) Conversion of obligations into registered...transfer agent or the issuer for conversion of the obligation into...

  7. 24 CFR 811.110 - Refunding of obligations issued to finance Section 8 projects.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...false Refunding of obligations issued to finance Section 8 projects. 811.110 Section...110 Refunding of obligations issued to finance Section 8 projects. (a) This...savings and, if necessary, HUD will finance in refunding bond debt service...

  8. 24 CFR 811.110 - Refunding of obligations issued to finance Section 8 projects.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ...false Refunding of obligations issued to finance Section 8 projects. 811.110 Section...110 Refunding of obligations issued to finance Section 8 projects. (a) This...savings and, if necessary, HUD will finance in refunding bond debt service...

  9. 24 CFR 811.110 - Refunding of obligations issued to finance Section 8 projects.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ...false Refunding of obligations issued to finance Section 8 projects. 811.110 Section...110 Refunding of obligations issued to finance Section 8 projects. (a) This...savings and, if necessary, HUD will finance in refunding bond debt service...

  10. 24 CFR 811.110 - Refunding of obligations issued to finance Section 8 projects.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ...false Refunding of obligations issued to finance Section 8 projects. 811.110 Section...110 Refunding of obligations issued to finance Section 8 projects. (a) This...savings and, if necessary, HUD will finance in refunding bond debt service...

  11. 24 CFR 811.110 - Refunding of obligations issued to finance Section 8 projects.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...false Refunding of obligations issued to finance Section 8 projects. 811.110 Section...110 Refunding of obligations issued to finance Section 8 projects. (a) This...savings and, if necessary, HUD will finance in refunding bond debt service...

  12. 31 CFR 103.52 - Photographic or other reproductions of Government obligations.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ...2010-07-01 false Photographic or other reproductions of Government obligations. 103... § 103.52 Photographic or other reproductions of Government obligations. Nothing...authorize the microfilming or other reproduction of (a) Currency or other...

  13. 31 CFR 1010.940 - Photographic or other reproductions of Government obligations.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ...2011-07-01 false Photographic or other reproductions of Government obligations. 1010...1010.940 Photographic or other reproductions of Government obligations. Nothing...authorize the microfilming or other reproduction of: (a) Currency or other...

  14. 47 CFR 14.61 - Obligations with respect to internet browsers built into mobile phones.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... false Obligations with respect to internet browsers built into mobile phones...EQUIPMENT BY PEOPLE WITH DISABILITIES Internet Browsers Built Into Telephones Used With...14.61 Obligations with respect to internet browsers built into mobile phones....

  15. 47 CFR 14.61 - Obligations with respect to internet browsers built into mobile phones.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... false Obligations with respect to internet browsers built into mobile phones...EQUIPMENT BY PEOPLE WITH DISABILITIES Internet Browsers Built Into Telephones Used With...14.61 Obligations with respect to internet browsers built into mobile phones....

  16. 42 CFR 64a.105 - What are the conditions of obligated service?

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ...105 Section 64a.105 Public Health PUBLIC HEALTH SERVICE, DEPARTMENT OF HEALTH AND HUMAN SERVICES FELLOWSHIPS, INTERNSHIPS, TRAINING OBLIGATED SERVICE FOR MENTAL HEALTH TRAINEESHIPS § 64a.105 What are the conditions of obligated...

  17. 76 FR 36482 - Affirmative Action and Nondiscrimination Obligations of Contractors and Subcontractors Regarding...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-06-22

    ...Action and Nondiscrimination Obligations of Contractors and Subcontractors Regarding Protected Veterans AGENCY: Office of Federal...Action and Nondiscrimination Obligations of Contractors and Subcontractors Regarding Protected Veterans'' (76 FR 23358)....

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    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-04-26

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    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-02-10

    ...Action and Nondiscrimination Obligations of Contractors and Subcontractors Regarding Individuals With Disabilities AGENCY: Office...Action and Nondiscrimination Obligations of Contractors and Subcontractors Regarding Individuals with Disabilities'' (76 FR...

  20. 76 FR 77055 - Affirmative Action and Nondiscrimination Obligations of Contractors and Subcontractors Regarding...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-12-09

    ...Non-discrimination Obligations of Contractors and Subcontractors Regarding Individuals With Disabilities...Nondiscrimination Obligations of Contractors and Subcontractors Regarding Individuals With Disabilities...by covered Federal contractors and subcontractors against individuals on the basis...

  1. 12 CFR 1511.5 - Obligations of Funding Corporation; no adverse claims.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ...2010-01-01 false Obligations of Funding Corporation; no adverse claims. ...DEPARTMENT OF THE TREASURY RESOLUTION FUNDING CORPORATION BOOK-ENTRY PROCEDURE § 1511.5 Obligations of Funding Corporation; no adverse...

  2. 40 CFR 80.1107 - How is the Renewable Volume Obligation calculated?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ...2011-07-01 2011-07-01 false How is the Renewable Volume Obligation calculated? 80.1107...REGULATION OF FUELS AND FUEL ADDITIVES Renewable Fuel Standard § 80.1107 How is the Renewable Volume Obligation calculated?...

  3. 40 CFR 80.1106 - To whom does the Renewable Volume Obligation apply?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ...2011-07-01 false To whom does the Renewable Volume Obligation apply? 80.1106...REGULATION OF FUELS AND FUEL ADDITIVES Renewable Fuel Standard § 80.1106 To whom does the Renewable Volume Obligation apply?...

  4. 40 CFR 80.1107 - How is the Renewable Volume Obligation calculated?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ...2010-07-01 2010-07-01 false How is the Renewable Volume Obligation calculated? 80.1107...REGULATION OF FUELS AND FUEL ADDITIVES Renewable Fuel Standard § 80.1107 How is the Renewable Volume Obligation calculated?...

  5. 40 CFR 80.1106 - To whom does the Renewable Volume Obligation apply?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ...2010-07-01 false To whom does the Renewable Volume Obligation apply? 80.1106...REGULATION OF FUELS AND FUEL ADDITIVES Renewable Fuel Standard § 80.1106 To whom does the Renewable Volume Obligation apply?...

  6. 32 CFR 220.2 - Statutory obligation of third party payer to pay.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ...COLLECTION FROM THIRD PARTY PAYERS OF REASONABLE CHARGES FOR HEALTHCARE SERVICES § 220.2 Statutory obligation of third party...obligation to pay the United States the reasonable charges for healthcare services provided in or through any facility of...

  7. 40 CFR 152.97 - Rights and obligations of data submitters.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ...2010-07-01 false Rights and obligations of data submitters. 152.97 Section 152...PROCEDURES Procedures To Ensure Protection of Data Submitters' Rights § 152.97 Rights and obligations of data submitters. (a) Right to be...

  8. 78 FR 63276 - Interim Policy, FAA Review of Solar Energy System Projects on Federally Obligated Airports

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-10-23

    ...Interim Policy, FAA Review of Solar Energy System Projects on Federally Obligated...obligated airports to construct solar energy systems on airport property. FAA...measuring ocular impact of proposed solar energy systems which are effective...

  9. Sixth Northwest Conservation and Electric Power Plan Chapter 13: Bonneville's Obligations

    E-print Network

    's balancing area. Its obligations to provide flexibility for wind-power balancing also are driven by its obligations under NERC standards as the host balancing authority for wind-power resources that are meeting

  10. 31 CFR 1010.311 - Filing obligations for reports of transactions in currency.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ...obligations for reports of transactions in currency. 1010.311 Section 1010.311...obligations for reports of transactions in currency. Each financial institution other...each deposit, withdrawal, exchange of currency or other payment or transfer,...

  11. 31 CFR 1010.311 - Filing obligations for reports of transactions in currency.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ...obligations for reports of transactions in currency. 1010.311 Section 1010.311...obligations for reports of transactions in currency. Each financial institution other...each deposit, withdrawal, exchange of currency or other payment or transfer,...

  12. 31 CFR 1010.311 - Filing obligations for reports of transactions in currency.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ...obligations for reports of transactions in currency. 1010.311 Section 1010.311...obligations for reports of transactions in currency. Each financial institution other...each deposit, withdrawal, exchange of currency or other payment or transfer,...

  13. 31 CFR 1010.311 - Filing obligations for reports of transactions in currency.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ...obligations for reports of transactions in currency. 1010.311 Section 1010.311...obligations for reports of transactions in currency. Each financial institution other...each deposit, withdrawal, exchange of currency or other payment or transfer,...

  14. 40 CFR 80.1407 - How are the Renewable Volume Obligations calculated?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ...following formulas: (1) Cellulosic biofuel. RVOCB,i = (RFStdCB,i ...Renewable Volume Obligation for cellulosic biofuel for an obligated party for calendar year...RFStdCB,i = The standard for cellulosic biofuel for calendar year i, determined by...

  15. 40 CFR 80.1407 - How are the Renewable Volume Obligations calculated?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ...following formulas: (1) Cellulosic biofuel. RVOCB,i = (RFStdCB,i ...Renewable Volume Obligation for cellulosic biofuel for an obligated party for calendar year...RFStdCB,i = The standard for cellulosic biofuel for calendar year i, determined by...

  16. 32 CFR 220.2 - Statutory obligation of third party payer to pay.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ...COLLECTION FROM THIRD PARTY PAYERS OF REASONABLE CHARGES FOR HEALTHCARE SERVICES § 220.2 Statutory obligation of third party...obligation to pay the United States the reasonable charges for healthcare services provided in or through any facility of...

  17. 32 CFR 220.2 - Statutory obligation of third party payer to pay.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ...COLLECTION FROM THIRD PARTY PAYERS OF REASONABLE CHARGES FOR HEALTHCARE SERVICES § 220.2 Statutory obligation of third party...obligation to pay the United States the reasonable charges for healthcare services provided in or through any facility of...

  18. 32 CFR 220.2 - Statutory obligation of third party payer to pay.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ...COLLECTION FROM THIRD PARTY PAYERS OF REASONABLE CHARGES FOR HEALTHCARE SERVICES § 220.2 Statutory obligation of third party...obligation to pay the United States the reasonable charges for healthcare services provided in or through any facility of...

  19. 32 CFR 220.2 - Statutory obligation of third party payer to pay.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ...COLLECTION FROM THIRD PARTY PAYERS OF REASONABLE CHARGES FOR HEALTHCARE SERVICES § 220.2 Statutory obligation of third party...obligation to pay the United States the reasonable charges for healthcare services provided in or through any facility of...

  20. 7 CFR 984.66 - Assistance of the Board in meeting reserve obligation.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ...false Assistance of the Board in meeting reserve obligation. 984.66 Section 984...CALIFORNIA Order Regulating Handling Reserve Walnuts § 984.66 Assistance of the Board in meeting reserve obligation. The Board may assist...

  1. 7 CFR 984.66 - Assistance of the Board in meeting reserve obligation.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ...false Assistance of the Board in meeting reserve obligation. 984.66 Section 984...CALIFORNIA Order Regulating Handling Reserve Walnuts § 984.66 Assistance of the Board in meeting reserve obligation. The Board may assist...

  2. 40 CFR 80.1407 - How are the Renewable Volume Obligations calculated?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ...following formulas: (1) Cellulosic biofuel. RVOCB,i = (RFStdCB,i ...Renewable Volume Obligation for cellulosic biofuel for an obligated party for calendar year...RFStdCB,i = The standard for cellulosic biofuel for calendar year i, determined by...

  3. 40 CFR 80.1407 - How are the Renewable Volume Obligations calculated?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ...following formulas: (1) Cellulosic biofuel. RVOCB,i = (RFStdCB,i ...Renewable Volume Obligation for cellulosic biofuel for an obligated party for calendar year...RFStdCB,i = The standard for cellulosic biofuel for calendar year i, determined by...

  4. 40 CFR 80.1407 - How are the Renewable Volume Obligations calculated?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ...following formulas: (1) Cellulosic biofuel. RVOCB,i = (RFStdCB,i ...Renewable Volume Obligation for cellulosic biofuel for an obligated party for calendar year...RFStdCB,i = The standard for cellulosic biofuel for calendar year i, determined by...

  5. 76 FR 7975 - Commodity Pool Operators and Commodity Trading Advisors: Amendments to Compliance Obligations

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-02-11

    ...Pool Operators and Commodity Trading Advisors: Amendments to Compliance Obligations...Pool Operators and Commodity Trading Advisors: Amendments to Compliance Obligations...Pool Operators and Commodity Trading Advisors. The Commission is proposing a new...

  6. 77 FR 11251 - Commodity Pool Operators and Commodity Trading Advisors: Compliance Obligations

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-02-24

    ...Pool Operators and Commodity Trading Advisors: Compliance Obligations; Harmonization...Pool Operators and Commodity Trading Advisors: Compliance Obligations AGENCY: Commodity...Pool Operators and Commodity Trading Advisors. The Commission is also adopting...

  7. 28 CFR 45.10 - Procedures to promote compliance with crime victims' rights obligations.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... Procedures to promote compliance with crime victims' rights obligations. 45.10... Procedures to promote compliance with crime victims' rights obligations. (a...that relate to protection of the rights of crime victims. See 18 U.S.C. 3771....

  8. 28 CFR 45.10 - Procedures to promote compliance with crime victims' rights obligations.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... Procedures to promote compliance with crime victims' rights obligations. 45.10... Procedures to promote compliance with crime victims' rights obligations. (a...that relate to protection of the rights of crime victims. See 18 U.S.C. 3771....

  9. 16 CFR 437.2 - The obligation to furnish written documents.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ...false The obligation to furnish written documents. 437.2 Section 437.2 Commercial...437.2 The obligation to furnish written documents. In connection with the offer for sale, sale, or promotion of a business...

  10. 16 CFR 437.2 - The obligation to furnish written documents.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ...false The obligation to furnish written documents. 437.2 Section 437.2 Commercial...437.2 The obligation to furnish written documents. In connection with the offer for sale, sale, or promotion of a business...

  11. Origins, Diversity, and Diversification of the Native Hawaiian Leafhoppers (Hemiptera: Cicadellidae: Nesophrosyne) and Their Obligate Endosymbionts

    E-print Network

    O'Grady, Patrick M.

    Origins, Diversity, and Diversification of the Native Hawaiian Leafhoppers (Hemiptera: Cicadellidae Leafhoppers (Hemiptera: Cicadellidae: Nesophrosyne) and Their Obligate Endosymbionts Copyright 2012 By Gordon Leafhoppers (Hemiptera: Cicadellidae: Nesophrosyne) and Their Obligate Endosymbionts by Gordon Morse Bennett

  12. The Complete Genome of Teredinibacter turnerae T7901: An Intracellular Endosymbiont of Marine Wood-Boring Bivalves (Shipworms)

    PubMed Central

    Yang, Joyce C.; Madupu, Ramana; Durkin, A. Scott; Ekborg, Nathan A.; Pedamallu, Chandra S.; Hostetler, Jessica B.; Radune, Diana; Toms, Bradley S.; Henrissat, Bernard; Coutinho, Pedro M.; Schwarz, Sandra; Field, Lauren; Trindade-Silva, Amaro E.; Soares, Carlos A. G.; Elshahawi, Sherif; Hanora, Amro; Schmidt, Eric W.; Haygood, Margo G.; Posfai, Janos; Benner, Jack; Madinger, Catherine; Nove, John; Anton, Brian; Chaudhary, Kshitiz; Foster, Jeremy; Holman, Alex; Kumar, Sanjay; Lessard, Philip A.; Luyten, Yvette A.; Slatko, Barton; Wood, Nicole; Wu, Bo; Teplitski, Max; Mougous, Joseph D.; Ward, Naomi; Eisen, Jonathan A.; Badger, Jonathan H.; Distel, Daniel L.

    2009-01-01

    Here we report the complete genome sequence of Teredinibacter turnerae T7901. T. turnerae is a marine gamma proteobacterium that occurs as an intracellular endosymbiont in the gills of wood-boring marine bivalves of the family Teredinidae (shipworms). This species is the sole cultivated member of an endosymbiotic consortium thought to provide the host with enzymes, including cellulases and nitrogenase, critical for digestion of wood and supplementation of the host's nitrogen-deficient diet. T. turnerae is closely related to the free-living marine polysaccharide degrading bacterium Saccharophagus degradans str. 2–40 and to as yet uncultivated endosymbionts with which it coexists in shipworm cells. Like S. degradans, the T. turnerae genome encodes a large number of enzymes predicted to be involved in complex polysaccharide degradation (>100). However, unlike S. degradans, which degrades a broad spectrum (>10 classes) of complex plant, fungal and algal polysaccharides, T. turnerae primarily encodes enzymes associated with deconstruction of terrestrial woody plant material. Also unlike S. degradans and many other eubacteria, T. turnerae dedicates a large proportion of its genome to genes predicted to function in secondary metabolism. Despite its intracellular niche, the T. turnerae genome lacks many features associated with obligate intracellular existence (e.g. reduced genome size, reduced %G+C, loss of genes of core metabolism) and displays evidence of adaptations common to free-living bacteria (e.g. defense against bacteriophage infection). These results suggest that T. turnerae is likely a facultative intracellular ensosymbiont whose niche presently includes, or recently included, free-living existence. As such, the T. turnerae genome provides insights into the range of genomic adaptations associated with intracellular endosymbiosis as well as enzymatic mechanisms relevant to the recycling of plant materials in marine environments and the production of cellulose-derived biofuels. PMID:19568419

  13. 48 CFR 252.232-7007 - Limitation of Government's obligation.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... accordingly. (e) If, solely by reason of failure of the Government to allot additional funds, by the dates... 48 Federal Acquisition Regulations System 3 2011-10-01 2011-10-01 false Limitation of Government's... of Provisions And Clauses 252.232-7007 Limitation of Government's obligation. As prescribed in...

  14. 48 CFR 252.232-7007 - Limitation of Government's obligation.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... accordingly. (e) If, solely by reason of failure of the Government to allot additional funds, by the dates... 48 Federal Acquisition Regulations System 3 2010-10-01 2010-10-01 false Limitation of Government's... of Provisions And Clauses 252.232-7007 Limitation of Government's obligation. As prescribed in...

  15. Asset retirement obligations: a reporting concern for healthcare facilities.

    PubMed

    Berg, Gary G; Bayes, Paul E; Morgan, Robert G

    2008-11-01

    FASB statements and SEC guidelines give direction as to how healthcare organizations should account for their asset retirement obligations (AROs) where environmental issues are concerned. A key consideration is that current costs associated with environmental problems, such as encapsulating asbestos, are to be accounted for as part of an asset's cost and depreciated over the asset's remaining life. PMID:18990844

  16. 7 CFR 1488.12 - Coverage of bank obligations.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 10 2010-01-01 2010-01-01 false Coverage of bank obligations. 1488.12 Section 1488.12 Agriculture Regulations of the Department of Agriculture (Continued) COMMODITY CREDIT CORPORATION, DEPARTMENT OF AGRICULTURE LOANS, PURCHASES, AND OTHER OPERATIONS FINANCING OF SALES OF AGRICULTURAL COMMODITIES Financing of Export Sales...

  17. 18 CFR 154.1 - Application; Obligation to file.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... to file. 154.1 Section 154.1 Conservation of Power and Water Resources FEDERAL ENERGY REGULATORY... Provisions and Conditions § 154.1 Application; Obligation to file. (a) The provisions of this part apply to filings pursuant to section 4 of the Natural Gas Act. (b) Every natural gas company must file with...

  18. 18 CFR 154.1 - Application; Obligation to file.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... to file. 154.1 Section 154.1 Conservation of Power and Water Resources FEDERAL ENERGY REGULATORY... Provisions and Conditions § 154.1 Application; Obligation to file. (a) The provisions of this part apply to filings pursuant to section 4 of the Natural Gas Act. (b) Every natural gas company must file with...

  19. 18 CFR 154.1 - Application; Obligation to file.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... to file. 154.1 Section 154.1 Conservation of Power and Water Resources FEDERAL ENERGY REGULATORY... Provisions and Conditions § 154.1 Application; Obligation to file. (a) The provisions of this part apply to filings pursuant to section 4 of the Natural Gas Act. (b) Every natural gas company must file with...

  20. 47 CFR 76.309 - Customer service obligations.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 47 Telecommunication 4 2013-10-01 2013-10-01 false Customer service obligations. 76.309 Section 76.309 Telecommunication FEDERAL COMMUNICATIONS COMMISSION (CONTINUED) BROADCAST RADIO SERVICES MULTICHANNEL VIDEO AND CABLE TELEVISION SERVICE General Operating Requirements § 76.309 Customer...

  1. 47 CFR 76.309 - Customer service obligations.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 47 Telecommunication 4 2011-10-01 2011-10-01 false Customer service obligations. 76.309 Section 76.309 Telecommunication FEDERAL COMMUNICATIONS COMMISSION (CONTINUED) BROADCAST RADIO SERVICES MULTICHANNEL VIDEO AND CABLE TELEVISION SERVICE General Operating Requirements § 76.309 Customer...

  2. 47 CFR 76.309 - Customer service obligations.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 47 Telecommunication 4 2012-10-01 2012-10-01 false Customer service obligations. 76.309 Section 76.309 Telecommunication FEDERAL COMMUNICATIONS COMMISSION (CONTINUED) BROADCAST RADIO SERVICES MULTICHANNEL VIDEO AND CABLE TELEVISION SERVICE General Operating Requirements § 76.309 Customer...

  3. 47 CFR 76.309 - Customer service obligations.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 47 Telecommunication 4 2010-10-01 2010-10-01 false Customer service obligations. 76.309 Section 76.309 Telecommunication FEDERAL COMMUNICATIONS COMMISSION (CONTINUED) BROADCAST RADIO SERVICES MULTICHANNEL VIDEO AND CABLE TELEVISION SERVICE General Operating Requirements § 76.309 Customer...

  4. 7 CFR 1948.92 - Grant approval and fund obligation.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 13 2014-01-01 2013-01-01 true Grant approval and fund obligation. 1948.92 Section 1948.92 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS-COOPERATIVE SERVICE, RURAL UTILITIES SERVICE, AND FARM SERVICE AGENCY, DEPARTMENT OF AGRICULTURE (CONTINUED) PROGRAM REGULATIONS...

  5. 7 CFR 1416.705 - Obligations of a participant.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... ASSISTANCE PROGRAMS 2005 Hurricane Tree Assistance Program § 1416.705 Obligations of a participant. (a) Eligible producers must execute all required documents and complete the 2005 Hurricane TAP funded practice... becomes ineligible for all or part of a 2005 Hurricane TAP benefit, the person and successor shall...

  6. 24 CFR 291.565 - Continuing obligations after purchase.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... purchase. 291.565 Section 291.565 Housing and Urban Development Regulations Relating to Housing and Urban... Neighbor Next Door Sales Program § 291.565 Continuing obligations after purchase. To remain in compliance.../her sole residence, the home purchased through the GNND Sales Program; and (b) Certify initially...

  7. 24 CFR 291.565 - Continuing obligations after purchase.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... purchase. 291.565 Section 291.565 Housing and Urban Development Regulations Relating to Housing and Urban... Neighbor Next Door Sales Program § 291.565 Continuing obligations after purchase. To remain in compliance.../her sole residence, the home purchased through the GNND Sales Program; and (b) Certify initially...

  8. 7 CFR 1488.12 - Coverage of bank obligations.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 10 2011-01-01 2011-01-01 false Coverage of bank obligations. 1488.12 Section 1488.12 Agriculture Regulations of the Department of Agriculture (Continued) COMMODITY CREDIT CORPORATION, DEPARTMENT... Financing of Export Sales of Agricultural Commodities From Private Stocks Under CCC Export Credit...

  9. 31 CFR 223.18 - Performance of agency obligations.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 31 Money and Finance: Treasury 2 2010-07-01 2010-07-01 false Performance of agency obligations. 223.18 Section 223.18 Money and Finance: Treasury Regulations Relating to Money and Finance (Continued) FISCAL SERVICE, DEPARTMENT OF THE TREASURY FINANCIAL MANAGEMENT SERVICE SURETY COMPANIES DOING...

  10. 31 CFR 223.18 - Performance of agency obligations.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 31 Money and Finance:Treasury 2 2011-07-01 2011-07-01 false Performance of agency obligations. 223.18 Section 223.18 Money and Finance: Treasury Regulations Relating to Money and Finance (Continued) FISCAL SERVICE, DEPARTMENT OF THE TREASURY FINANCIAL MANAGEMENT SERVICE SURETY COMPANIES DOING...

  11. Life-Long Learning and Social Responsibility Obligations

    ERIC Educational Resources Information Center

    Mayes, Robin

    2013-01-01

    The literature affirms that widespread lapses in corporate social responsibility obligations (unethical behaviors) have periodically brought about extensive forfeitures of economic wealth and countless job losses leaving the world economy in recession or depression. Put forth as a resolution to unemployment issues the academic literature champions…

  12. Obama states obligation to act on climate change

    NASA Astrophysics Data System (ADS)

    Showstack, Randy

    2012-11-01

    Obama states obligation to act on climate change Noting increased global temperatures, Arctic ice melt, and severe weather events, President Barack Obama said that climate change is real and called for a conversation across the country to determine what can be done about it.

  13. 47 CFR 27.1168 - Triggering a Reimbursement Obligation.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ...apply the following test to determine when an AWS entity or MSS/ATC entity has triggered...cost-sharing obligation and therefore must pay an AWS relocator, MSS relocator (including MSS...initially co-channel with the licensed AWS band(s) of the AWS entity or the...

  14. 47 CFR 27.1184 - Triggering a reimbursement obligation.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ...the following test to determine when an AWS entity has triggered a cost-sharing obligation and therefore must pay an AWS relocator of a BRS system in accordance...initially co-channel with the licensed AWS band(s) of the AWS entity;...

  15. 47 CFR 27.1184 - Triggering a reimbursement obligation.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ...the following test to determine when an AWS entity has triggered a cost-sharing obligation and therefore must pay an AWS relocator of a BRS system in accordance...initially co-channel with the licensed AWS band(s) of the AWS entity;...

  16. 47 CFR 27.1168 - Triggering a Reimbursement Obligation.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ...apply the following test to determine when an AWS entity or MSS/ATC entity has triggered...cost-sharing obligation and therefore must pay an AWS relocator, MSS relocator (including MSS...initially co-channel with the licensed AWS band(s) of the AWS entity or the...

  17. 47 CFR 27.1168 - Triggering a Reimbursement Obligation.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ...apply the following test to determine when an AWS entity or MSS/ATC entity has triggered...cost-sharing obligation and therefore must pay an AWS relocator, MSS relocator (including MSS...initially co-channel with the licensed AWS band(s) of the AWS entity or the...

  18. 47 CFR 27.1168 - Triggering a Reimbursement Obligation.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ...the following test to determine when an AWS entity has triggered a cost-sharing obligation and therefore must pay an AWS relocator, MSS relocator, or a voluntarily...initially co-channel with the licensed AWS band(s) of the AWS entity or the...

  19. 47 CFR 27.1184 - Triggering a reimbursement obligation.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ...the following test to determine when an AWS entity has triggered a cost-sharing obligation and therefore must pay an AWS relocator of a BRS system in accordance...initially co-channel with the licensed AWS band(s) of the AWS entity;...

  20. 47 CFR 27.1184 - Triggering a reimbursement obligation.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ...the following test to determine when an AWS entity has triggered a cost-sharing obligation and therefore must pay an AWS relocator of a BRS system in accordance...initially co-channel with the licensed AWS band(s) of the AWS entity;...

  1. 47 CFR 27.1168 - Triggering a Reimbursement Obligation.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ...the following test to determine when an AWS entity has triggered a cost-sharing obligation and therefore must pay an AWS relocator, MSS relocator, or a voluntarily...initially co-channel with the licensed AWS band(s) of the AWS entity or the...

  2. 47 CFR 27.1184 - Triggering a reimbursement obligation.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ...the following test to determine when an AWS entity has triggered a cost-sharing obligation and therefore must pay an AWS relocator of a BRS system in accordance...initially co-channel with the licensed AWS band(s) of the AWS entity;...

  3. 47 CFR 54.405 - Carrier obligation to offer Lifeline.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... (CONTINUED) UNIVERSAL SERVICE Universal Service Support for Low-Income Consumers § 54.405 Carrier obligation...-day time period. A carrier providing Lifeline service in a state that has dispute resolution... information must be reported by month and must be submitted to the Commission at the time an...

  4. 47 CFR 54.1006 - Public interest obligations.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ...) UNIVERSAL SERVICE Mobility Fund § 54.1006 Public interest obligations. (a) Deadline for construction—3G... to enable the use of real time applications, such as VoIP. (b) Deadline for construction—4G networks... low enough to enable the use of real time applications, such as VoIP. (c) Coverage test data....

  5. 47 CFR 54.1006 - Public interest obligations.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ...) UNIVERSAL SERVICE Mobility Fund § 54.1006 Public interest obligations. (a) Deadline for construction—3G... to enable the use of real time applications, such as VoIP. (b) Deadline for construction—4G networks... low enough to enable the use of real time applications, such as VoIP. (c) Coverage test data....

  6. 47 CFR 54.405 - Carrier obligation to offer Lifeline.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... (CONTINUED) UNIVERSAL SERVICE Universal Service Support for Low-Income Consumers § 54.405 Carrier obligation...-day time period. A carrier providing Lifeline service in a state that has dispute resolution... information must be reported by month and must be submitted to the Commission at the time an...

  7. 47 CFR 54.405 - Carrier obligation to offer Lifeline.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... (CONTINUED) UNIVERSAL SERVICE Universal Service Support for Low-Income Consumers § 54.405 Carrier obligation...-day time period. A carrier providing Lifeline service in a state that has dispute resolution... information must be reported by month and must be submitted to the Commission at the time an...

  8. 47 CFR 54.1006 - Public interest obligations.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ...) UNIVERSAL SERVICE Mobility Fund § 54.1006 Public interest obligations. (a) Deadline for construction—3G... to enable the use of real time applications, such as VoIP. (b) Deadline for construction—4G networks... low enough to enable the use of real time applications, such as VoIP. (c) Coverage test data....

  9. 21 CFR 26.75 - Suspension of recognition obligations.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... MUTUAL RECOGNITION OF PHARMACEUTICAL GOOD MANUFACTURING PRACTICE REPORTS, MEDICAL DEVICE QUALITY SYSTEM AUDIT REPORTS, AND CERTAIN MEDICAL DEVICE PRODUCT EVALUATION REPORTS: UNITED STATES AND THE EUROPEAN... obligations under subpart A or B of this part, in whole or in part, if: (a) A party suffers a loss of...

  10. 13 CFR 120.171 - Compliance with child support obligations.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 13 Business Credit and Assistance 1 2014-01-01 2014-01-01 false Compliance with child support... Compliance with child support obligations. Any holder of 50% or more of the ownership interest in the... to pay child support arising under: (a) An administrative order; (b) A court order; (c) A...

  11. 13 CFR 120.171 - Compliance with child support obligations.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 13 Business Credit and Assistance 1 2011-01-01 2011-01-01 false Compliance with child support... Compliance with child support obligations. Any holder of 50% or more of the ownership interest in the... to pay child support arising under: (a) An administrative order; (b) A court order; (c) A...

  12. 22 CFR 230.07 - Fiscal Agent obligations.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 22 Foreign Relations 1 2010-04-01 2010-04-01 false Fiscal Agent obligations. 230.07 Section 230.07 Foreign Relations AGENCY FOR INTERNATIONAL DEVELOPMENT ISRAEL LOAN GUARANTEES ISSUED UNDER THE EMERGENCY WARTIME SUPPLEMENTAL APPROPRIATIONS ACT OF 2003, PUB. L. 108-11-STANDARD TERMS AND CONDITIONS §...

  13. 22 CFR 230.07 - Fiscal Agent obligations.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 22 Foreign Relations 1 2013-04-01 2013-04-01 false Fiscal Agent obligations. 230.07 Section 230.07 Foreign Relations AGENCY FOR INTERNATIONAL DEVELOPMENT ISRAEL LOAN GUARANTEES ISSUED UNDER THE EMERGENCY WARTIME SUPPLEMENTAL APPROPRIATIONS ACT OF 2003, PUB. L. 108-11-STANDARD TERMS AND CONDITIONS §...

  14. 22 CFR 230.07 - Fiscal Agent obligations.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 22 Foreign Relations 1 2011-04-01 2011-04-01 false Fiscal Agent obligations. 230.07 Section 230.07 Foreign Relations AGENCY FOR INTERNATIONAL DEVELOPMENT ISRAEL LOAN GUARANTEES ISSUED UNDER THE EMERGENCY WARTIME SUPPLEMENTAL APPROPRIATIONS ACT OF 2003, PUB. L. 108-11-STANDARD TERMS AND CONDITIONS §...

  15. 22 CFR 230.07 - Fiscal Agent obligations.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 22 Foreign Relations 1 2012-04-01 2012-04-01 false Fiscal Agent obligations. 230.07 Section 230.07 Foreign Relations AGENCY FOR INTERNATIONAL DEVELOPMENT ISRAEL LOAN GUARANTEES ISSUED UNDER THE EMERGENCY WARTIME SUPPLEMENTAL APPROPRIATIONS ACT OF 2003, PUB. L. 108-11-STANDARD TERMS AND CONDITIONS §...

  16. 22 CFR 230.07 - Fiscal Agent obligations.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 22 Foreign Relations 1 2014-04-01 2014-04-01 false Fiscal Agent obligations. 230.07 Section 230.07 Foreign Relations AGENCY FOR INTERNATIONAL DEVELOPMENT ISRAEL LOAN GUARANTEES ISSUED UNDER THE EMERGENCY WARTIME SUPPLEMENTAL APPROPRIATIONS ACT OF 2003, PUB. L. 108-11-STANDARD TERMS AND CONDITIONS §...

  17. 7 CFR 760.507 - Obligations of a participant.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... AGRICULTURE SPECIAL PROGRAMS INDEMNITY PAYMENT PROGRAMS Tree Assistance Program § 760.507 Obligations of a participant. (a) Eligible orchardists and nursery tree growers must execute all required documents and... nursery tree growers must allow representatives of FSA to visit the site for the purposes of...

  18. 7 CFR 760.507 - Obligations of a participant.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... AGRICULTURE SPECIAL PROGRAMS INDEMNITY PAYMENT PROGRAMS Tree Assistance Program § 760.507 Obligations of a participant. (a) Eligible orchardists and nursery tree growers must execute all required documents and... nursery tree growers must allow representatives of FSA to visit the site for the purposes of...

  19. 7 CFR 760.507 - Obligations of a participant.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... AGRICULTURE SPECIAL PROGRAMS INDEMNITY PAYMENT PROGRAMS Tree Assistance Program § 760.507 Obligations of a participant. (a) Eligible orchardists and nursery tree growers must execute all required documents and... nursery tree growers must allow representatives of FSA to visit the site for the purposes of...

  20. 7 CFR 760.507 - Obligations of a participant.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... AGRICULTURE SPECIAL PROGRAMS INDEMNITY PAYMENT PROGRAMS Tree Assistance Program § 760.507 Obligations of a participant. (a) Eligible orchardists and nursery tree growers must execute all required documents and... nursery tree growers must allow representatives of FSA to visit the site for the purposes of...

  1. 34 CFR 686.40 - Documenting the service obligation.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... EDUCATION (TEACH) GRANT PROGRAM Service and Repayment Obligations § 686.40 Documenting the service... ceasing enrollment in a program of study for which a TEACH Grant was received, the grant recipient must..., science, a foreign language, bilingual education, English language acquisition, special education, or as...

  2. 25 CFR 226.9 - Rental and drilling obligations.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 25 Indians 1 2013-04-01 2013-04-01 false Rental and drilling obligations. 226.9 Section 226.9... RESERVATION LANDS FOR OIL AND GAS MINING Leasing Procedure, Rental and Royalty § 226.9 Rental and drilling... in the lease terms, or 12 months from the date the Superintendent consents to drilling on...

  3. 43 CFR 3162.2 - Drilling, producing, and drainage obligations.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 43 Public Lands: Interior 2 2014-10-01 2014-10-01 false Drilling, producing, and drainage obligations. 3162.2 Section 3162.2 Public Lands: Interior Regulations Relating to Public Lands (Continued... OPERATIONS Requirements for Operating Rights Owners and Operators § 3162.2 Drilling, producing, and...

  4. 25 CFR 226.9 - Rental and drilling obligations.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 25 Indians 1 2010-04-01 2010-04-01 false Rental and drilling obligations. 226.9 Section 226.9... RESERVATION LANDS FOR OIL AND GAS MINING Leasing Procedure, Rental and Royalty § 226.9 Rental and drilling... in the lease terms, or 12 months from the date the Superintendent consents to drilling on...

  5. 43 CFR 3162.2 - Drilling, producing, and drainage obligations.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 43 Public Lands: Interior 2 2013-10-01 2013-10-01 false Drilling, producing, and drainage obligations. 3162.2 Section 3162.2 Public Lands: Interior Regulations Relating to Public Lands (Continued... OPERATIONS Requirements for Operating Rights Owners and Operators § 3162.2 Drilling, producing, and...

  6. 25 CFR 226.9 - Rental and drilling obligations.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 25 Indians 1 2014-04-01 2014-04-01 false Rental and drilling obligations. 226.9 Section 226.9... RESERVATION LANDS FOR OIL AND GAS MINING Leasing Procedure, Rental and Royalty § 226.9 Rental and drilling... in the lease terms, or 12 months from the date the Superintendent consents to drilling on...

  7. 25 CFR 226.9 - Rental and drilling obligations.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 25 Indians 1 2012-04-01 2011-04-01 true Rental and drilling obligations. 226.9 Section 226.9... RESERVATION LANDS FOR OIL AND GAS MINING Leasing Procedure, Rental and Royalty § 226.9 Rental and drilling... in the lease terms, or 12 months from the date the Superintendent consents to drilling on...

  8. 43 CFR 3162.2 - Drilling, producing, and drainage obligations.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 43 Public Lands: Interior 2 2012-10-01 2012-10-01 false Drilling, producing, and drainage obligations. 3162.2 Section 3162.2 Public Lands: Interior Regulations Relating to Public Lands (Continued... OPERATIONS Requirements for Operating Rights Owners and Operators § 3162.2 Drilling, producing, and...

  9. 25 CFR 226.9 - Rental and drilling obligations.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 25 Indians 1 2011-04-01 2011-04-01 false Rental and drilling obligations. 226.9 Section 226.9... RESERVATION LANDS FOR OIL AND GAS MINING Leasing Procedure, Rental and Royalty § 226.9 Rental and drilling... in the lease terms, or 12 months from the date the Superintendent consents to drilling on...

  10. 43 CFR 3162.2 - Drilling, producing, and drainage obligations.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 43 Public Lands: Interior 2 2011-10-01 2011-10-01 false Drilling, producing, and drainage obligations. 3162.2 Section 3162.2 Public Lands: Interior Regulations Relating to Public Lands (Continued... OPERATIONS Requirements for Operating Rights Owners and Operators § 3162.2 Drilling, producing, and...

  11. 28 CFR 811.3 - Notice of obligation to register.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... COLUMBIA SEX OFFENDER REGISTRATION § 811.3 Notice of obligation to register. (a) Sex offenders may be... Sex Offender Registration Act of 1999 (D.C. Official Code sections 22-4003, 4005, 4007) (relating to.... 14071(b)(1) (notice under federal law standards for state sex offender registration programs). (b)...

  12. 32 CFR 901.25 - Obligation of cadet appointment.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 901.25 National Defense Department of Defense (Continued) DEPARTMENT OF THE AIR FORCE MILITARY TRAINING AND SCHOOLS APPOINTMENT TO THE UNITED STATES AIR FORCE ACADEMY Nomination Procedures and Requirements § 901.25 Obligation of cadet appointment. (a) A cadet who enters the Air Force Academy...

  13. 31 CFR 225.5 - Pledge of definitive Government obligations.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... (Continued) FISCAL SERVICE, DEPARTMENT OF THE TREASURY FINANCIAL MANAGEMENT SERVICE ACCEPTANCE OF BONDS... the bond official under this part shall be made at the risk and expense of the obligor. Upon receipt of definitive Government obligations, the bond official will issue the obligor a receipt. (c) Risk...

  14. 7 CFR 1450.105 - Obligations of participant.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 10 2011-01-01 2011-01-01 false Obligations of participant. 1450.105 Section 1450.105 Agriculture Regulations of the Department of Agriculture (Continued) COMMODITY CREDIT CORPORATION, DEPARTMENT OF AGRICULTURE LOANS, PURCHASES, AND OTHER OPERATIONS BIOMASS CROP ASSISTANCE PROGRAM (BCAP)...

  15. 7 CFR 1450.105 - Obligations of participant.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 10 2012-01-01 2012-01-01 false Obligations of participant. 1450.105 Section 1450.105 Agriculture Regulations of the Department of Agriculture (Continued) COMMODITY CREDIT CORPORATION, DEPARTMENT OF AGRICULTURE LOANS, PURCHASES, AND OTHER OPERATIONS BIOMASS CROP ASSISTANCE PROGRAM (BCAP)...

  16. 7 CFR 1450.105 - Obligations of participant.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 10 2013-01-01 2013-01-01 false Obligations of participant. 1450.105 Section 1450.105 Agriculture Regulations of the Department of Agriculture (Continued) COMMODITY CREDIT CORPORATION, DEPARTMENT OF AGRICULTURE LOANS, PURCHASES, AND OTHER OPERATIONS BIOMASS CROP ASSISTANCE PROGRAM (BCAP)...

  17. 7 CFR 1450.105 - Obligations of participant.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 10 2014-01-01 2014-01-01 false Obligations of participant. 1450.105 Section 1450.105 Agriculture Regulations of the Department of Agriculture (Continued) COMMODITY CREDIT CORPORATION, DEPARTMENT OF AGRICULTURE LOANS, PURCHASES, AND OTHER OPERATIONS BIOMASS CROP ASSISTANCE PROGRAM (BCAP)...

  18. 22 CFR 211.5 - Obligations of cooperating sponsor.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... USE IN DISASTER RELIEF, ECONOMIC DEVELOPMENT AND OTHER ASSISTANCE § 211.5 Obligations of cooperating... abortions as a method of family planning or to motivate or coerce any person to practice abortions. (5.... Except in the case of emergency or disaster situations, the donation of commodities furnished for...

  19. 22 CFR 211.5 - Obligations of cooperating sponsor.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... USE IN DISASTER RELIEF, ECONOMIC DEVELOPMENT AND OTHER ASSISTANCE § 211.5 Obligations of cooperating... abortions as a method of family planning or to motivate or coerce any person to practice abortions. (5.... Except in the case of emergency or disaster situations, the donation of commodities furnished for...

  20. 22 CFR 211.5 - Obligations of cooperating sponsor.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... USE IN DISASTER RELIEF, ECONOMIC DEVELOPMENT AND OTHER ASSISTANCE § 211.5 Obligations of cooperating... abortions as a method of family planning or to motivate or coerce any person to practice abortions. (5.... Except in the case of emergency or disaster situations, the donation of commodities furnished for...