Implant Dent 22:71–76PubMedCrossRef 22. Kuroshima buy TPCA-1 S, Go VA, Yamashita J (2012) Increased numbers of nonattached osteoclasts after long-term zoledronic acid therapy in mice. Endocrinology 153:17–28PubMedCrossRef 23. Yamashita J, Koi K, Yang DY, McCauley LK (2011)

Effect of zoledronate on oral wound healing in rats. Clin Cancer Res 17:1405–1414PubMedCentralPubMedCrossRef 24. Enlow DH (1966) Osteocyte necrosis in normal bone. J Dent Res 45:213PubMedCrossRef 25. Bonnet N, Lesclous P, Saffar JL, Ferrari S (2013) Zoledronate effects on systemic and jaw osteopenias in ovariectomized periostin-deficient mice. PLoS One 8:e58726 26. McDonald MM, Dulai S, Godfrey C, Amanat N, Sztynda T, Little DG (2008) Bolus or weekly zoledronic acid administration does not delay endochondral fracture repair but

weekly dosing enhances delays in hard callus remodeling. Bone 43:653–662PubMedCrossRef 27. Peter CP, Cook WO, Nunamaker DM, Provost MT, Seedor JG, Rodan GA (1996) Effect of alendronate on fracture healing and bone remodeling in dogs. J Orthop Res 14:74–79PubMedCrossRef 28. Allen MR, Chu TM, Ruggiero SL (2013) Absence BTK inhibitor of exposed bone following dental extraction in beagle dogs treated with 9 months of high-dose zoledronic acid combined with dexamethasone. J Oral Maxillofac Surg 71:1017–1026PubMedCrossRef 29. Watts NB, Diab DL (2010) Long-term use of bisphosphonates in osteoporosis. J Clin Endocrinol Metab 95:1555–1565PubMedCrossRef 30. McMillan MD Tau-protein kinase (1975) An ultrastructural study of the relationship of oral bacteria to the epithelium of healing tooth extraction wounds. Arch

Oral Biol 20:815–822PubMedCrossRef 31. Ravanelli A, J, K (2006) Cranifofacial development. Lippincott Williams & Wikins, Philadelphia 32. Eames BF, Helms JA (2004) Conserved molecular program regulating cranial and MRT67307 concentration appendicular skeletogenesis. Dev Dyn 231:4–13PubMedCrossRef 33. Aghaloo TL, Kang B, Sung EC, Shoff M, Ronconi M, Gotcher JE, Bezouglaia O, Dry SM, Tetradis S (2011) Periodontal disease and bisphosphonates induce osteonecrosis of the jaws in the rat. J Bone Miner Res 26:1871–1882PubMedCentralPubMedCrossRef 34. Aguirre JI, Akhter MP, Kimmel DB, Pingel JE, Williams A, Jorgensen M, Kesavalu L, Wronski TJ (2012) Oncologic doses of zoledronic acid induce osteonecrosis of the jaw-like lesions in rice rats (Oryzomys palustris) with periodontitis. J Bone Miner Res 27:2130–2143PubMedCentralPubMedCrossRef 35. Lopez-Jornet P, Camacho-Alonso F, Martinez-Canovas A, Molina-Minano F, Gomez-Garcia F, Vicente-Ortega V (2011) Perioperative antibiotic regimen in rats treated with pamidronate plus dexamethasone and subjected to dental extraction: a study of the changes in the jaws. J Oral Maxillofac Surg 69:2488–2493PubMedCrossRef 36.

Structure of mature biofilms The quantitative representation of the used species was most convincing when biofilms were grown in iHS medium. T. selleck compound denticola established in high numbers and the biofilms showed the best stability during the following staining procedures. Therefore, structural analysis was focused on these biofilms. CLSM analyses of FISH stained biofilms enabled us to determine all 10 species used in the model and locate their position in the biofilms. The top layer (approximately 30 μm from the biofilm surface) and basal layer (approximately 50 μm from the disc surface) of the biofilms showed clear structural differences and a fluent transition between these layers was observed. Selleck RG-7388 Biofilms grown in mFUM4 showed

a dominance of F. nucleatum and streptococci in the basal layer (Figure 5A). In biofilms grown in iHS, however, F. nucleatum was detectable by FISH only in the top layer as dispersed cells, while streptococci were very abundant throughout the whole biofilm (Figure 5B). Aggregations of streptococci were often mixed with V. dispar in the whole biofilm except in the top layer, where V. dispar occurred as compact microcolonies (Figure 6). In biofilms grown in mFUM4, which had a lower thickness,

this growth pattern of V. dispar was observed throughout the biofilm (Figure 5A). P. intermedia was found predominantly in the lower half of the biofilms Selleck MK5108 forming microcolonies with diameters of about 50 μm on average (Figure 7A). T. forsythia was found mainly in the top layer of the biofilm, while none were detected in the lower half of the biofilms (Figure 7A). T. denticola grew loosely in the top layer alongside with P. gingivalis, which displayed the highest density in close proximity to T. denticola accumulations (Figure 7B). A. oris appeared as loose EPS-embedded microcolonies located in the upper half of the biofilms (Figure 8A). Campylobacter rectus was dispersed throughout the biofilm and did not form own microcolonies, but showed higher density in the top layer of the biofilm

(Figure 8B). Figure 5 Biofilms grown for 64.5 h in or mFUM4- (A) or iHS medium(B). FISH staining of a fixed biofilm; Endonuclease the biofilm base in the side views is directed towards the top view. (A) red: F. nucleatum, white: V. dispar, green: non-hybridised cells, DNA staining (YoPro-1 + Sytox), blue: EPS. (B) cyan: streptococci, red: F. nucleatum, green: non-hybridised cells, DNA staining (YoPro-1 + Sytox). Figures show a representative area of one disc. Scale bars: 20 μm. Figure 6 Biofilms grown for 64.5 h in iHS medium. FISH staining of a fixed biofilm; the biofilm base in the side views is directed towards the top view. Cyan: V. dispar, green: non-hybridised cells, DNA staining (YoPro-1 + Sytox). Arrows: Microcolonies of V. dispar. Shown is a representative area of one disc. Scale bar: 30 μm. Figure 7 3D-reconstructions of a 146 x 146 μm section of biofilms grown for 64.5 h in iHS medium. FISH staining of a fixed biofilm. P. gingivalis and T.

braziliensis (day 1) and were given 250 μg of the respective antibody every 3 days for 3 weeks thereafter. Statistical analyses The data are reported as the mean ± SEM and are representative of two or three independent experiments. The means between different groups were compared by the analysis of variance (ANOVA) followed by the Tukey test for

unpaired values. P<0.05 was considered to be statistically significant. Results Kinetics of the SGE effect in the recruitment of leucocytes to the site of inoculation in BALB/c mice We analyzed the accumulation of leukocytes in the dermis after 0, 6, 12, 24 and 48 hours post-intradermal inoculation of Lutzomyia longipalpis PF-573228 mouse salivary gland extract (SGE) (0.5 pair of glands/mouse) into the ears of BALB/c mice. SGE induced neutrophils (GR1+MHC-II–) cell recruitment 6 hours this website post-inoculation, which persisted for 48 hours. CD4+ T cells, CD8+ T cells and CD4+CD25+ T cells appeared 12 hours post-inoculation and persisted during all period analyzed. Macrophages (F4/80+CD11c- MHC-II+) cell accumulation was observed 12 hours after inoculation, and dendritic cells (CD11b+CD11c+MHC-II+) levels did not change (Figure  1A). Figure 1 Kinetics of the inflammatory infiltrate induced check details by Lutzomyia longipalpis saliva at the site

of inoculation. BALB/c mice were inoculated intradermally within the ear dermis with half of the salivary gland extract (SGE) generated from the two salivary glands diluted in 10 μl of PBS (A) or a injection with PBS only (B). The leucocytes from three mouse ears/group were obtained at 0, 6, 12, 24 and 48 h after inoculation,

and different populations were identified using flow cytometry. The data showed represent the mean ± SEM and are representative of three independent experiments (n = 3). *P < 0.05 compared to 0 hours (naive). To determine that the leukocyte migration is SGE-specific and not due damage inflicted by the needle injection, the kinect of leucocyte migration after similar amounts of PBS (10 μL) inoculated into ears of mice was performed. As showed, the amounts of dendritic cells, neutrophils, macrophages in PBS-inoculated mice was similar in all time points analyzed and was comparable that those recovered Quisqualic acid from naïve ears mice (Figure  1B), confirming the specificity of SGE in the leukocyte recruitment. Inflammatory infiltrate after one or three inocula of SGE Next, we determined whether saliva promotes or protects against leishmaniasis. First, we compared the inflammatory infiltrate after different injections of SGE. BALB/c mice received one or three intradermal ear injections of SGE, and the emigrated leucocytes were analyzed. As a control group, BALB/c mice were inoculated with PBS (time 0). Our results show that the SGE-1X group had an increased recruitment of different subtypes analyzed: CD4+ T cells, CD8+ T cells, CD4+CD25+ cells, macrophage and neutrophil (Figure  2).

PubMedCrossRef 8. Beersma MF, Dirven K, Van Dam AP, Templeton KE, Claas EC, Goossens H: Evaluation of 12 commercial tests and the complement fixation

test for MLN2238 clinical trial Mycoplasma pneumoniae specific immunoglobulin G (IgG) and IgM antibodies, with PCR used as the “”gold standard”". J Clin Microbiol 2005, 43:2277–2285.PubMedCrossRef 9. Dorigo-Zetsma JW, Zaat SA, Wertheim-van Dillen PM, Spanjaard L, Rijntjes , Van Waveren G, Jensen JS, Angulo AF, Dankert J: Comparison of PCR, culture, and serological tests for diagnosis of Mycoplasma pneumoniae respiratory tract infection in children. J Clin Microbiol 1999, 37:14–17.PubMed 10. Suni J, Vainionpaa R, Tuuminen T: Multicenter evaluation of the novel enzyme immunoassay based on P1-enriched protein GS-4997 for the detection of Mycoplasma pneumoniae infection. J Microbiol Methods 2001, 47:65–71.PubMedCrossRef 11. Tuuminen T, Suni J, Kleemola M, Jacobs E: Improved sensitivity

and specificity learn more of enzyme immunoassays with P1-adhesin enriched antigen to detect acute Mycoplasma pneumoniae infection. J Microbiol Methods 2001, 44:27–37.PubMedCrossRef 12. Csango PA, Pedersen JE, Hess RD: Comparison of four Mycoplasma pneumoniae IgM-, IgG- and IgA-specific enzyme immunoassays in blood donors and patients. Clin Microbiol Infect 2004, 10:1094–1098.PubMedCrossRef 13. Chaudhry R, Nisar N, Hora B, Chirasani SR, Malhotra P: Expression and immunological characterization of the carboxy-terminal region of the P1 adhesin protein of Mycoplasma pneumoniae . J Clin Microbiol 2005, 43:321–325.PubMedCrossRef 14. Dallo SF, Su CJ, Horton JR, Baseman JB: Identification of P1 gene domain containing epitope(s) mediating Mycoplasma pneumoniae cytoadherence. J Exp Med 1988, 167:718–723.PubMedCrossRef 15. Drasbek M, Nielsen PK, Persson K, Birkelund S, Christiansen G: Immune response to Mycoplasma pneumoniae P1 and P116 in patients with atypical pneumonia analyzed by ELISA. BMC Microbiol 2004, 4:7–17.PubMedCrossRef 16. Dumke R, Schurwanz N, Jacobs E: Characterisation of subtype- and variant specific antigen regions of the P1 adhesin of Mycoplasma pneumoniae . Int J Med Microbiol 2008,

298:483–491.PubMedCrossRef click here 17. Jacobs E, Bennewitz A, Bredt W: Reaction pattern of human anti- Mycoplasma pneumoniae antibodies in enzyme-linked immunosorbent assays and immunoblotting. J Clin Microbiol 1986, 23:517–522.PubMed 18. Duffy MF, Whithear KG, Noormohammadi AH, Markham PF, Catton M, Leydon J, Browning GF: Indirect enzyme-linked immunosorbent assay for detection of immunoglobulin G reactive with a recombinant protein expressed from the gene encoding the 116-kilodalton protein of Mycoplasma pneumoniae . J Clin Microbiol 1999, 37:1024–1029.PubMed 19. Varshney AK, Chaudhry KR, Kabra SK, Malhotra P: Cloning, expression, and immunological characterization of the P30 protein of Mycoplasma pneumoniae . Clin Vaccine Immunol 2008, 15:215–220.PubMedCrossRef 20.

Up-regulated transport genes have been shown or predicted to be involved in the uptake of L-aminoacids or peptides (aapJ, aapQ, aapP, oppB, oppC, SMc00140, SMc01597, SMc02259, SMb21572, SMb20605), branched-chain aminoacids (livH, livM, livG, livF, livK), uracil/uridine (SMc01823, SMc01824, SMc01825, SMc01827), sugar amines (SMb21151) or other complex N substrates such as the polyamines

spermidine and putrescine (SMc01966, SMc01965, SMc01963). Consequently, loss of hfq also resulted in the up-regulation of an important set of genes likely related to the utilization or modification of amino acids and other N compounds. The transcripts corresponding to the 3 genes specifying the glycine cleavage system, gcvP, gcvH and gcvT (M values 2.06, 2.02 and 3.32, respectively), and to SMc01930 (M value 3.26) encoding a putative methylmalonyl-CoA INCB28060 ic50 epimerase likely operating in the catabolism of branched-chain amino acids were particularly over-represented in the mutant. The proteomic analysis of the other hfq mutant (2011-3.4) used selleck chemicals llc in this study identified periplasmic solute

binding proteins of ABC transporters and metabolic enzymes as the predominant sets of polypeptides which accumulation in the cell was altered by disruption of the hfq gene by the insertion of pK18mobsacB (Fig. 3, lower circle graphs). Down-regulated transport proteins are all involved in the uptake of different sugars; myo-inositol (IbpA), mannose/xylose/glucose (AraA), fructose (FrcB) and α-glucosides (AglE). Accordingly, several enzymes of the central carbon metabolism were also less abundant in the mutant: a putative myo-inositol catabolic protein (IolE), a predicted malonic semialdehyde oxidative decarboxylase (IolD) and a probable acetyl-CoA synthetase (AcsA1). Conversely, the transporters overproduced by the 2011-3.4 mutant are all related to the import of N substrates such as peptides oxyclozanide (DppA1 and DppA2), leucine (LivK), L-amino acids (AapJ and AapP), other aminoacids (SMc02259), glycine betaine (SMc02378) or choline (ChoX). Other up-regulated proteins as a result of the hfq mutation include metabolic

enzymes such as ornithine cyclodeaminase (Ocd), a probable arginase (ArgI1), a putative adenosylhomocysteinase (AhcY) and a phosphoenol pyruvate carboxykinase (PckA). Ocd and ArgI1 selleck inhibitor catalyze enzymatic reactions of the urea cycle whereas AhcY is involved in the metabolism of sulphur-containing aminoacids. PckA catalyzes the conversion of oxalacetate into phospho-enol pyruvate, thus initiating the gluconeogenic pathway. In summary, transcriptomics and proteomics independently suggest that in both S. meliloti hfq knock-out mutants metabolism is biased towards the gluconeogenesis pathway so that growth of free-living bacteria is mainly supported by the utilization of amino acids rather than primary carbon substrates as energy sources. Loss of Hfq affects S.

g. inoculation) post infection. Each https://www.selleckchem.com/products/DAPT-GSI-IX.html sample was analyzed in triplicate and the analysis was repeated at least twice. The CFU of the sample was expressed as the average of the values obtained. The concentrations of bacteria were recorded as CFU/ml of organ homogenate. The limit of bacteria detection in the organ homogenates was 10 CFU/ml. In vitrosynthesis and secretion of the tagged SPI-1 proteins under different cultured conditions After being ingested from contaminated food or polluted water,Salmonellawill encounter a series of extreme environmental changes such as acidity in the

stomach, hypoxia, hyperosmolarity, and other conditions learn more such as fermentation in the gut [22–24]. The expression of bacterial genes including those of SPI-1 is expected to be regulated to allow bacteria to adapt to new environments and to prepare for the invasion of the intestinal epithelium. To investigate the synthesis and secretion of the SPI-1 proteins, each of the tagged strains was grown under five different conditions that resembled the early stages of its natural infection, and the expression of the tagged proteins was

studied. (A) Expression in rich media LB broth Western analyses were carried out to detect the expression of the ATM/ATR cancer tagged proteins with an anti-FLAG antibody (Figure2Aand2B), using the expression of bacterial DnaK protein as the internal control (Figure2C). Normalization of samples was also carried out by loading total protein extracted from the same CFU (e.g. 5 × 107CFU) of bacteria in each lane. SPI-1 proteins PrgI, SopE2, SpaO, SptP, SipB, and SipA were detected inSalmonellacultured in LB broth (Figure2B). Furthermore, SpoE2, SptP, SipB, and SipA but not PrgI or SpaO Glycogen branching enzyme were detected in the culture supernatant (Figure2A, data not

shown). These results are consistent with the previous observations that SpaO and PrgI are the structural components of the needle complex [5]. Figure 2 Western analyses of the synthesis (B) and secretion (A) of the tagged proteins from bacterial strains T-prgI (13 KD)(lane 1), T-spoE2 (29 KD) (lane 2), T-spaO (36 KD) (lane 3), T-sptP (62 KD) (lane 4), T-sipB (65 KD) (lane 5), and T-sipA (76 KD) (lane 6). The expression of bacterial DnaK was used as the internal control (C). Protein samples were separated in SDS-polyacrylamide gels and reacted with antibodies against the FLAG sequence (A-B) and DnaK (C). Each lane was loaded with material from 5 × 107CFU bacteria. The molecular masses of some of the proteins in the PageRuler protein size markers (Fermentas) are shown and given in kiloDaltons (KD). (B) Expression under different pH conditions Acidity in the stomach is the first stressSalmonellameets after being ingested orally. Because the environment in the intestine is relatively basic,Salmonellawill encounter an increase in pH after it reaches the intestine.

Hence, similar to other complex bacterial communities and the intestinal microflora [16] and the bacterial vaginosis microflora [6, 7] in particular, much C646 of the bacterial diversity of the neovaginal microflora – consisting of 8.6 cultivable species on average – remains uncharacterized through culture techniques. Still, using culture, twelve possibly novel species, designated TSW Genotypes A to L, were detected. Specific assays were applied to detect fungal species in the neovagina, as well as to assess the presence of the index species in bacterial vaginosis. While Candida albicans was

not seen on Gram stain, six women were found to harbour some fungal species, of which three remain unidentified. No firm conclusions can be drawn on the virtual absence of Candida albicans, however it may be acknowledged that in biological women Candida species are AZD4547 frequently encountered on the vaginal mucosa and that their colonisation of the vagina is stimulated

by estradiol. It is interesting to notice that the neovagina is colonized by largely the same intestinal species as the vagina in the absence of lactobacilli (some two thirds of transsexual women (64.0%) having evidence of colonisation with G. vaginalis, A. vaginae, or both, and over 80% of transsexual women harbouring M. curtisii) and thus that the same type of colonisation selleck chemical occurs, regardless the type of epithelium, vaginal mucosa of skin. The BVAB species 1 to 3, belonging Palbociclib molecular weight to the phylum Clostridiales, and BVAB2 in particular, were found to be highly specific for BV in biological women [17, 18]. These were however not found in the neovaginal specimens, although it must be noted that the incidence

of these species in the vaginal microflora of female patients of our hospital was quite low as well (unpublished data). It is difficult to establish to what extent a bacterial vaginosis-like condition is present among these transsexual women. In a recent case report, a transsexual woman was diagnosed with bacterial vaginosis based on a Hay-Ison score of 2 (intermediate microflora) [19], although microscopy revealed numerous white blood cells and spirochaete-like organisms consistent with our observations. As a matter of fact, based on the criterion of a departure from a normal lactobacilli-dominated microflora, all patients in our study would qualify as having bacterial vaginosis-like microflora. We failed to document any association between specific bacteria and vaginal complaints – except for the unexplained and possibly spurious correlation between the presence of Mobiluncus and pain during sexual activity. It should be noted that also among biological women vaginitis symptoms are rarely associated with a single micro-organism.

pertussis DNA using primers with the restriction sites BamHI (PrnProF-BamHI) and NdeI (PRNProR-NdeI). The plasmid pSKPD25FpPRN3 was cut with BamHI and NdeI to generate a fragment which had lost the FHA promoter. The PRN promoter AZD5363 in vivo was ligated in its place. After transformation into E. coli and verification by restriction analysis, the resulting plasmid was designated as pSKPD25PRN3 (Figure 5C). The plasmid was

cut with NotI and inserted into pSS4245 cut with the same enzyme. The resulting construct, pSSPD2prn was transferred into E. coli SM10 to conduct the allelic exchange. The resulting B. pertussis strain was designated as Bp-WWE. Integration of the prn gene at its designated position was confirmed by PCR with the primers that specifically bind only to the upstream 5′ (5′FPD2-int and PRNProR-NdeI primers), 3′ (PRNF-int and 3′RPD2-int primers) downstream flanking regions, and inside the prn gene. PT, FHA and PRN expression in shake flask culture The Bp-WWC, Bp-WWD and Bp-WWE strains were grown in shake flasks with 100 mL MSS medium supplemented with methylated β-cyclodextrin (1 g/l) at 35°C with shaking speed of 200 rpm. After 32-48 h of growth, the culture supernatants were collected and assayed by ELISA to quantify the PT and FHA expression level. As PRN releasing from its membrane-bound precursor is the result of an imprecise cleavage by unidentified proteases

[34], PRN expression was determined by Western blot with densitometric buy MI-503 analysis to evaluate the integrity of the antigen. Nutlin-3 in vivo MTMR9 This assay was conducted both on the clarified culture supernatant and the cell extract obtained by heating cell suspension in isotonic buffer (10 mM Tris-HCl pH 8.0, 150 mM NaCl, 0.002% NaN3, and 1 mM PMSF) at 60°C for 30 min and the supernatant was collected after centrifugation

at 10,000 × g, 4°C for 30 min. ELISA assay for PT and FHA Purified rabbit polyclonal antibodies against PT or FHA (NLAC, Thailand) with the dilution of 1:1000 in carbonate/bicarbonate buffer (pH 9.6) were coated in 96-well plates (NUNC Maxisorp, Denmark) for 100 μL per well and incubated overnight at 4°C. After 3 time-washing with phosphate-buffered saline pH 7.4 containing 0.1% Tween 20 (PBST), blocking was performed using 100 μL per well of 3% bovine serum albumin (BSA)-PBST then incubated at 37°C for 1 h. After discarding the blocking buffer and washing, dilutions of the standard PT, FHA or samples were loaded and incubated at 37°C for 1 h. Then, anti-PT mouse monoclonal antibody (Abcam, USA) at 1:30,000 dilution or anti-FHA mouse monoclonal antibody (NIBSC, UK) at 1:10,000 dilution in blocking buffer was added and incubated under the same conditions. After washing the wells for three times with PBST, 100 μL of rabbit anti-mouse (H + L) IgG-HRP conjugate (Abcam, USA) in blocking buffer at 1:10,000 dilution was used as secondary antibody and incubated for 37°C for 1 h.

Acta Biochim Biophys Sin 2007, 38:79–88.CrossRef 47. Li Y, Hu Y, Fu W, Xia B, Jin C: Solution structure of the bacterial chemotaxis adaptor protein CheW from Escherichia

coli . Biochem Biophys Res Commun 2007, 360:863–867.PubMedCrossRef 48. Porter SL, Warren AV, Martin AC, Armitage JP: The third chemotaxis locus of Rhodobacter sphaeroides CFTRinh-172 price is essential for chemotaxis. Mol Microbiol 2002, 46:1081–1094.PubMedCrossRef 49. Stock AM, Robinson VL, Goudreau PN: Idasanutlin price Two-component signal transduction. Ann Rev Biochem 2000, 69:183–215.PubMedCrossRef 50. Jiang ZY, Bauer CE: Component of the Rhodospirillum centenum photosensory apparatus with structural and functional similarity to methyl-accepting chemotaxis protein receptors. J Bacteriol 2001, 183:171–177.PubMedCrossRef 51. Maniatis T, Fritsch EF, Sambrook J: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor: Cold Spring Harbor Press; 1982. 52. Miroux B, Walker JE: Over-production of proteins in Escherichia coli : mutant hosts that allow synthesis of some membrane proteins and globular proteins at high levels. J Mol Biol 1996, 260:289–298.PubMedCrossRef 53. Abouhamad WN, Manson M, Gibson MM, Higgins CF: Peptide transport and chemotaxis in Escherichia coli

and Salmonella typhimurium : characterization of the dipeptide permease see more (Dpp) and the dipeptide-binding protein. Mol Microbiol 1991, 5:1035–1047.PubMedCrossRef 54. Tabor S, Richardson CC: A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes. Proc Natl Acad SciUSA 1985, 82:1074–1078.CrossRef 55. Guzman LM, Belin D, Carson MJ, Beckwith J: Tight regulation, modulation, and high-level expression by vectors containing the arabinose pBAD promoter. J Bacteriol 1995, 177:4121–4130.PubMed 56. Miller J: Experiments in Molecular Genetics. Cold Spring Harbor Laboratory; 1972. 57. Laemmli UK: Cleavage of structural

proteins during the assembly of the head of bacteriophage T4. Nature 1970, 227:680–685.PubMedCrossRef 58. Figurski DH, Helinski DR: Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid Dichloromethane dehalogenase function provided in trans . Proc Natl Acad Sci USA 1979, 76:1648–1652.PubMedCrossRef 59. Willetts N: Conjugation. Methods Microbiol 1984, 17:33–59.CrossRef 60. Kiefer D, Hu X, Dalbey R, Kuhn A: Negatively charged amino acid residues play an active role in orienting the Sec-independent Pf3 coat protein in the Escherichia coli inner membrane. EMBO J 1997, 16:2197–2204.PubMedCrossRef 61. T D, Kuhn A: Hydrophobic forces drive spontaneous membrane insertion of the bacteriophage Pf3 coat protein without topological control. EMBO J 1999, 18:6299–6306.CrossRef 62. Tartoff KD, Hobbs CA: Improved media for growing plasmid and cosmid clones. Bethesda Res Lab Focus 1987, 9:12. 63. Schkölziger S: Klonierung und Expression des ppr -Gens aus Rhodospirillum centenum . In Diploma-thesis. University of Hohenheim, Institute of Microbiology; 2000.

ichthiosmia CECT 4486T 132 – - 122 104 116 110 81 19 102 Environment, Surface water

– NA, Germany, 1986   A. veronii bv. sobria LMG 13067 144 – - 132 115 126 121 8 93 12 Non-human, Frog I Connecticut, USA, NA   A. caviae (n=34) BVH16 9 1 B 9 8 9 9 3 8 8 Human, Respiratory tract C Rambouillet, Fr, 2006   BVH57 43 1 B 43 8 9 9 3 32 8 Human, Blood I Versailles, Fr, 2006   BVH63 47 6 F 12 10 43 41 3 10 41 Human, selleck chemicals llc Blood I Macon, Fr, 2006   BVH84 47 6 F 12 10 43 41 3 10

41 Human, Stool I Aix en Provence, Fr, 2006   BVH98 72 – F 12 10 64 60 37 10 41 Human, Wound I Brest, Fr, NA   ADV118 79 6 F 72 10 43 8 3 10 41 Human, Wound I Montpellier, Fr, 2009   ADV121 81 – F 74 10 43 8 3 3 63 Human, Stool ND Montpellier, Fr, 2009   BVH48 34 2 C 34 10 32 32 27 26 32 Human, Vagina C Monceau les mines, Fr, 2006   A. caviae CCUG 48892 175 2 C 34 10 32 32 27 3 32 Environment, Water   Uppsala, Sweden, 2004   BVH19 11 – C 11 10 3 11 3 10 10 Human, Vagina C Villeneuve sur Lot, Fr, 2006   BVH81 61 – C 34 10 3 11 3 26 32 Human, Stool C Aix en Provence, Fr, CX-5461 in vitro 2006   BVH66 50 – C 34 10 46 44 37 26 32 Human, Wound I Martinique Island, Fr, 2006   BVH55 41 3 C 41 10 39 12 3 26 32 Human, Stool I Saint-Denis, Fr, 2006   BVH87 64 3 C 59 10 39 12 3 26 32 Human, Stool I Aix en Provence, Fr, 2006   BVH4 3 – - 3 3 3 3 3 3 3 Human, Wound I Cahors, Fr, 2006

  BVH15 8 – - 8 7 8 8 6 7 7 Human, Blood I Grasse, Fr, 2006   BVH20 12 – - 12 10 11 12 3 8 11 Human, Protein kinase N1 Stool I Gonesse, Fr, 2006   BVH51 37 – - 37 32 35 35 29 28 35 Human, Blood I Monaco, Fr, 2006   BVH52 38 – - 38 33 36 36 30 29 36 Human, Blood I Monaco, Fr, 2006   BVH67 51 – - 49 32 47 45 3 8 35 Human, Stool ND Martinique Island, Fr, NA   BVH85 62 – - 57 48 55 11 3 40 8 Human, Stool I Aix en Provence, Fr, 2006   BVH86 63 – - 58 49 56 53 43 41 51 Human, Stool C Aix en Provence, Fr, 2006   BVH100 73 – - 67 56 65 61 50 26 58 Human, Wound ND Brest, Fr, ND   ADV106 77 – - 70 59 68 64 52 50 35 Human, Stool ND Montpellier, Fr, 2008   ADV124 82 – - 75 62 71 67 3 53 64 Human, Stool ND Montpellier, Fr, 2009   AK223 98 – - 91 74 86 81 3 8 77 Environment, Waste water HSP inhibitor drugs treatment lagoon – Montracol, Fr, 2006   AK229 101 – - 34 77 89 84 37 3 78 Environment, Waste water treatment lagoon – Montracol, Fr, 2006   AK231 102 – - 94 78 90 85 63 26 32 Environment, Waste water treatment lagoon – Montracol, Fr, 2006   AK234 104 – - 96 10 92 87 65 66 80 Environment, Waste water treatment lagoon – Montracol, Fr, 2006   AK245 115 – - 105 88 100 11 70 71 88 Environment, Water lake – Annecy, Fr, 1998   A.