Numbers at branch-points are percentages of 1000 bootstrap resamp

Numbers at branch-points are percentages of 1000 bootstrap resamplings

that support the topology of the tree. Sequencing was carried out on the fliC gene of sixteen randomly selected isolates of R. pickettii, and the type strain of R. insidiosa. The phylogenetic analysis of the fliC gene can be seen in Figure 2b, with the isolates divided into two branches with B. cepacia as an out-group. The isolates identified as R. insidiosa in-group two grouped together with groups three BIBW2992 cell line and four. These however were not supported by high bootstrapping values. Group 1 is made up of R. pickettii isolates from clinical and environmental sources with 97-100% similarity to the R. pickettii type strain. Group 2 is made up of R. insidiosa with 85% similarity to the R. pickettii type strain; Group 3 is made up of both R. insidiosa and R. pickettii with 86-87% similarity to the R. pickettii type strain and Group 4 is made up of the available sequenced R. pickettii strains with 87% similarity to the R. pickettii type strain. The division of the groups did not correlate to clinical or environmental association or on their isolation location. These Ricolinostat solubility dmso results indicate that there see more is variation in

the flagellin gene of R. pickettii. RAPD PCR results and analysis RAPD analysis was carried out using four different primers, three of which (P3, P15 and M13) have been shown to discriminate between selleckchem closely related strains of Ralstonia spp. including R. mannitolilytica and Cupriavidus pauculus [Ralstonia paucula] [47, 48]. The reproducibility of the RAPD method was tested by repeating the RAPD assays at least three times for each primer used (data not shown). The results revealed that apart from some variations in the band intensity, no significant differences were observed between the profiles

obtained, confirming the reproducibility of the method. Fifty-nine isolates of R. pickettii and R. insidiosa were characterised by RAPD analysis using all four primers and all isolates were placed into genotypes (Table 3). Percent similarities based on the Pearson correlation coefficients and clustering by the UPGMA method for these isolates using the OPA03U primer is presented in Figure 3a. Dendograms for the other primers (P3, P15 and M13) are presented in Additional File 2, Figure S1, S2 and S3. Fragments ranged from approximately 100 to 1800 bp for all primers. Clusters were distinguished at a similarity cut-off level of 80%. No major differentiation between the clinical, industrial, laboratory purified water and type strains could be observed, as these all fell into separate groups (Table 3) with each primer. For each of the primers there were a number of groups, with M13 there were twenty-one groups, OPA3OU there were 15 groups, P3 there were twenty-five groups and with primer P15 there were twenty-one groups.

J Clin

J Clin Endocrinol Metab 86:192–199PubMedCrossRef 12. Van Pottelbergh I, Goemaere S, Kaufman JM (2003) Bioavailable estradiol and an aromatase gene polymorphism are determinants of bone mineral density changes in men over 70 years of age. J Clin Endocrinol Metab 88:3075–3081PubMedCrossRef 13. Fink HA, Ewing SK, Ensrud KE, Barrett-Connor E, Taylor BC, Cauley JA, Orwoll ES (2006) Association of testosterone and estradiol deficiency with osteoporosis

and rapid bone loss in older men. J Clin Endocrinol Metab 91:3908–3915PubMedCrossRef 14. Khosla S, Melton LJ 3rd, Robb RA, Camp JJ, Atkinson EJ, Oberg AL, Rouleau PA, Riggs BL (2005) Relationship of volumetric BMD and structural parameters at different skeletal sites to sex steroid levels in men.

J Bone Miner Res 20:730–740PubMedCrossRef 15. Szulc P, Uusi-Rasi K, Claustrat B, Marchand F, Beck TJ, Delmas PD (2004) Role of sex steroids in the regulation of bone find more morphology in men. The MINOS study. Osteoporos Int 15:909–917PubMedCrossRef 16. Lauretani F, Bandinelli S, Russo CR, Maggio M, Di Iorio A, Cherubini A, Maggio D, Ceda GP, Valenti G, Guralnik JM, Ferrucci L (2006) Correlates of bone quality in older persons. Bone 39:915–921PubMedCrossRef 17. Bjornerem A, Ahmed LA, Joakimsen RM, Berntsen GK, Fonnebo V, Jorgensen L, Oian P, Seeman E, Straume B (2007) GF120918 mw A prospective study of sex steroids, sex hormone-binding globulin, and non-vertebral fractures in women and men: the Tromso Study. Eur J Endocrinol 157:119–125PubMedCrossRef 18. Bjornerem A, Emaus N, Berntsen GK, Joakimsen RM, Fonnebo V, Wilsgaard T, Oian P, Seeman E, Straume B (2007) Circulating sex steroids, sex hormone-binding globulin, and longitudinal changes in forearm bone mineral density in postmenopausal women and men: the tromso study. Calcif Tissue Int 81:65–72PubMedCrossRef 19. Goderie-Plomp HW, van der Klift M, de Ronde W, Hofman A, de Jong FH, Pols HAP (2004) Endogenous sex hormones, sex hormone-binding globulin, and the risk of incident vertebral fractures in elderly men and women: the Rotterdam study. J Clin Endocrinol Metab 89:3261–3269PubMedCrossRef many 20. Lee DM, O’Neill TW, Pye

SR, Silman AJ, Finn JD, Pendleton N, Tajar A, PCI32765 Bartfai G, Casanueva F, Forti G, Giwercman A, Huhtaniemi IT, Kula K, Punab M, Boonen S, Vanderschueren D, Wu FC (2009) The European Male Ageing Study (EMAS): design, methods and recruitment. Int J Androl 32:11–24PubMedCrossRef 21. Augat P, Reeb H, Claes L (1996) Prediction of fracture load at different skeletal sites by geometric properties of the cortical shell. J Bone Miner Res 11:1356–1363PubMedCrossRef 22. Schiessl H, Ferretti J, Tysarczyk-Niemeyer G, Willnecker J (1996) Noninvasive bone strength index as analyzed by peripheral quantitative computed tomography (pQCT). In: Schoenau E (ed) Paediatric osteology: new developments in diagnostics and therapy. Elsevier, Amsterdam, pp 141–146 23.

PubMedCrossRef 23 André T, Boni C, Mounedji-Boudiaf

L, N

PubMedCrossRef 23. André T, Boni C, Mounedji-Boudiaf

L, Navarro M, Tabernero J, Hickish T, Topham C, Zaninelli M, Clingan P, Bridgewater J, Tabah-Fisch I, de Gramont A: Oxaliplatin, fluorouracil, and leucovorin as adjuvant treatment for colon cancer. N Engl J Med 2004,350(23):2343–2351.PubMedCrossRef 24. Thorsteinsson MKL, Lund LR, Sørensen LT, Gerds TA, Jess P, Olsen J: Gene expression profiles in stage II and III colon cancer. Application of a 128-gene signature. Int J Colorectal Dis 2012,27(12):1579–1586.PubMedCrossRef 25. Smith JJ, Deane NG, Wu F, Merchant NB, Zhang B, Jiang A, Lu P, Johnson JC, Schmidt C, Bailey CE, Eschrich S, Kis C, Levy S, Washington MK, Heslin MJ, Coffey RJ, Yeatman TJ, Shyr Y, Beauchamp RD: Experimentally derived metastasis gene expression profile predicts recurrence and death in patients with colon cancer. Gastroenterology 2010,138(3):958–968.PubMedCentralPubMedCrossRef Selleckchem HM781-36B 26. Staub E, Groene J, Heinze M, Mennerich D, Roepcke S, Klaman I, Hinzmann B, Castanos-Velez E, Pilarsky C, Mann B, Brümmendorf T, Weber B, Buhr HJ, Rosenthal A: An expression module of WIPF1-coexpressed genes identifies patients with favorable prognosis in three tumor types. J Mol Med (Berl) 2009,87(6):633–644.CrossRef 27.

de Sousa E Melo F, Colak S, Buikhuisen J, Koster J, Cameron K, de Jong JH, Tuynman JB, Prasetyanti PR, Fessler E, van den Bergh SP, Rodermond H, Dekker E, van der selleck chemical Loos CM, Pals ST, van de Vijver MJ, Versteeg R, Richel DJ, Vermeulen L, Medema JP: Methylation of cancer-stem-cell-associated Wnt target genes predicts poor prognosis in colorectal cancer patients. Cell Stem Cell 2011,9(5):476–485.PubMedCrossRef 28. Reid JF, Gariboldi M, Sokolova V, Capobianco P, Lampis A, Perrone F, Signoroni S, Costa A, Leo E, Pilotti S, Pierotti MA: Integrative approach for prioritizing cancer genes in sporadic colon cancer. Genes Chromosomes Cancer 2009,48(11):953–962.PubMedCrossRef 29. Kaiser S, Park YK, Franklin JL, Halberg RB, Ribociclib Yu M, Jessen WJ, Freudenberg J, Chen X, Haigis K, Jegga AG, Kong S, Sakthivel B, Xu H, Reichling T, Azhar M, Boivin GP, Roberts RB, Bissahoyo AC, Gonzales F, Bloom GC, Eschrich S, Carter SL, Aronow JE, Kleimeyer J, Kleimeyer M, Ramaswamy V, Settle

SH, Boone B, Levy S, Graff JM, et al.: Transcriptional recapitulation and subversion of embryonic colon development by mouse colon tumor Selleck MM-102 models and human colon cancer. Genome Biol 2007,8(7):R131.PubMedCentralPubMedCrossRef 30. Gryfe R, Kim H, Hsieh ET, Aronson MD, Holowaty EJ, Bull SB, Redston M, Gallinger S: Tumor microsatellite instability and clinical outcome in young patients with colorectal cancer. N Engl J Med 2000,342(2):69–77.PubMedCrossRef 31. Jorissen RN, Lipton L, Gibbs P, Chapman M, Desai J, Jones IT, Yeatman TJ, East P, Tomlinson IP, Verspaget HW, Aaltonen LA, Kruhøffer M, Orntoft TF, Andersen CL, Sieber OM: DNA copy-number alterations underlie gene expression differences between microsatellite stable and unstable colorectal cancers.

Peripheral quantitative computed tomography Peripheral QCT measur

Peripheral quantitative computed tomography Peripheral QCT measurements of the non-dominant radius

were made in men recruited to the Manchester and Leuven centres using XCT-2000 scanners (Stratec, Pforzheim, Germany). At the distal (4%) site total and trabecular BMD (mg/cm3) and bone cross-sectional area (mm2) were measured (voxel size, 0.4 mm); the slice location at the 4% and 50% site was more distal in Leuven compared with Manchester; the reference line was placed at the distal Nec-1s mw border of the radial endplate in Leuven, in Manchester the line is placed to bisect the lateral border of the endplate these differences result in a scan site difference approximately 1–2 mm between centers. At the diaphysis (50% MGCD0103 clinical trial site, voxel size 0.6 mm), cortical BMD click here (mg/cm3), cortical BMC (mg/mm), total, cortical and medullary areas (mm2), cortical thickness (mm), stress strain index (SSI, mm3) and muscle cross-sectional area, as a proxy for muscle strength (CSMA, mm2), were measured. SSI provides a measure of a bone’s torsional strength [21, 22]. A detailed methodology for these measurements has been described previously [23]. For cross-calibration between Leuven and Manchester the European Forearm Phantom (EFP) was measured [24]; 10 repeat

measurements were taken in slices 1–4. There were no differences greater than precision error for trabecular, total and cortical BMD, BMC or cortical area. Therefore no cross-calibration was performed

between the two centres. These data and decisions were reviewed by Dr Klaus Engelke a CT expert from University of Erlangen, Germany and the scanner manufacturer Stratec Medizintechnik GmbH, Profzheim, Germany (Dr. Johannes Willnecker—personal communication). The short term precision of two repeat radius measurements with repositioning in adults were: Manchester (n = 22) Leuven (n = 40) trabecular BMD 1.27%, 1.42%; total BMD 2.1%, 1.3%; cortical BMD 0.77%, 0.71%; cortical area 2.4%, 1.3%; muscle area 3.7%, 1.1%. Manufacturer’s standard quality assurance procedures were followed in both Amylase centres. Sex hormone measurement A single-fasting morning (before 10.00 h) venous blood sample was obtained from all subjects. Serum was separated immediately after phlebotomy and stored at −80°C until assay at the end of the baseline study. Measurement of T and E2 were carried out by gas chromatography mass spectrometry as described in Labrie et al. [25, 26]. The lower limit of T quantitation was 0.17 nmol/L and E2 was 7.34 pmol/L. The coefficients of variation of T measurements were 2.9% within runs and 3.4% between runs, and for E2, were 3.5% within runs and 3.7% between runs. SHBG was measured by the Modular E170 platform electrochemiluminescence immunoassay (Roche Diagnostics, Mannheim, Germany) as previously described [27].

After additional solvent development, the contrast curve (Figure 

After additional solvent development, the contrast curve (Figure 1b) shows a mixed behavior, rather than a simple positive or negative tone behavior. At very low exposure doses, since the unexposed LCZ696 mw resist is soluble in pentyl acetate developer whereas electron beam exposure decomposed the resist to generate less soluble decomposition product, the resist exhibited a negative tone. At higher doses, on the one hand, the resist was increasingly decomposed and vaporized with increasing doses, which led to the tendency of positive tone; on the other hand, as the degree of decomposition increased, the decomposition product became less soluble

in the solvent developer, resulting in the tendency of negative tone after solvent development. As a consequence of those two competing trends, there exists a turning point exposure dose (approximately 1,200 μC/cm2) that gave a maximum remaining thickness. Such an exposure behavior can lead to complex structure as shown in Figure 2b, which is due to proximity exposure at the surrounding area beyond the directly exposed area. In fact, such kind of mixed exposure property is well known for a long time for PMMA that displays a positive tone at low doses and becomes a negative tone at approximately 10 times higher doses [21], which was also employed to

generate complex structures [22]. Though less known, another popular resist ZEP-520A actually also exhibits a mixed tone behavior just like PMMA [23]. However, unlike PMMA and ZEP for which the negative tone behavior JNK-IN-8 manufacturer appears only after roughly 10 times higher doses, for nitrocellulose, the negative tone behavior proceeds the positive tone, and the

dose ranges for the two tones have a large overlap and thus they are not clearly separated. E-beam working distance optimization using nitrocellulose resist Figure 3a illustrates the pattern design within the 1 mm × 1 mm writing field that consists of five identical wheel-structure array at the center and four corners, respectively, with the inset showing the wheel-structure array having exponentially increasing line doses from the upper left to the lower right wheel. A broad range of exposure dose is critical because Protein tyrosine phosphatase a relatively low dose is needed to reveal the high resolution capability when the beam is well focused, yet a high dose is essential to self-develop the resist to a certain visible depth when the beam is seriously enlarged. The wheel design is advantageous as it contains lines along various directions, which ensures that some lines (those roughly along the beam spot CH5424802 concentration elongation direction when there is severe astigmatism) would be adequately self-developed to become visible under SEM. Figure 3 CAD pattern design and structures exposed in nitrocellulose. (a) The CAD pattern design consisting of five identical wheel array structures (see right side for zoom-in view) at the 1 mm × 1 mm writing field center and four corners.

Enhancement of apoptosis in synchronized cell To determine whethe

Enhancement of Syk inhibitor apoptosis in synchronized cell To determine whether the improvement of HSV-tk gene transfer efficiency by cell synchronization resulted into an increased GCV-mediated cell death, we measured the level of cell apoptosis after GCV treatment using annexin V-FITC. The presence of apoptosis observed with annexin V labeling was confirmed by the DNA fragmentation method (Figure 5). Annexin V labeling

was increased in MTX-treated DHDK12 and HT29 cells transduced with HSV-tk gene and then treated for 72 hr by GCV. Figure 5 Internucleosomal Quisinostat DNA fragmentation induced by GCV. Lane 1 and lane 4 show DHDK12 cells and HT29 cells transduced with TG 9344 and treated for 96 hr with 20 μM GCV, respectively. Lane 3 and 5 show DHDK12 cells and HT29 cells transduced with TG 9344 after a 24 hr pretreatment with MTX and treated for 96 hr with 20 μM GCV, respectively. Lane 6 and 7 show DHDK12 cells and HT29 cells treated for 24 h with MTX, respectively. Lane 2 shows pBR 322 base pair size markers. Qualitative detection of DNA was achieved by ethidium bromide staining. In non-transduced cells, 5% of

MTX treated cells GS-1101 cost were labeled for annexin V-FITC after treatment by GCV (Figure 6A). This corresponds to the intrinsic toxicity of MTX. Figure 6 Induction of apoptosis. Untransduced DHDK12 cells (A) were treated with MTX, GCV or the combination of MTX plus GCV for 24 h. Transduced DHDK12 cells (B) and transduced HT29 cells (C) were treated for 24 hr with (filled Megestrol Acetate square) or without (open square) MTX. Cells were transduced with TG 9344 at the indicated times after MTX washout and 48 hr after transduction were treated with 20 μM GCV for 72

hr. Quantitative detection of apoptosis was determined by biparametric flow cytometry analysis of fluorescein labeled-annexin V cells and PI. Apoptotic cells were annexin V positive, PI negative. Data are expressed as the mean ± SE from at least three separate experiments. * P <.05 vs untreated cells The percentage of MTX-treated DHDK12 cells undergoing apoptosis (Annexin V+, PI-) was two fold higher after MTX withdrawal (46% vs. 23% in the untreated cell population). The difference was maximal in cells transduced 20 hr after MTX withdrawal (Figure 6B). In HT29 cells, the maximum percentage of MTX-treated cells undergoing apoptosis was 28% while it was 20% in untreated cells. The highest level of cell apoptosis was maximal 6 hr after MTX withdrawal (Figure 6C). Discussion The objective of this work was to improve the efficiency of retroviral transfer of the suicide gene HSV-tk in colon cancer cells. This aim was achieved through the pharmacological control of the target cells cell cycle. Our results are consistent with previous reports showing that retroviral-mediated gene transfer depends on the cell cycle of target cells.

Unpaired Student t test was used to compare ALL

Unpaired Student t test was used to compare ALL this website with Control group. Statistical significances are shown between groups only when p ≤ 0.05. Figure 4 Levels of PBX1 – 4 in healthy volunteers vs. patients with leukemia. Box plot graphics showing ΔCP values RG7420 taking ACTB (left panel) or RPL32 (right panel) as reference genes. The graphics display median (dark lines), 25‒75th percentile (boxes), interquartile ranges (whiskers), and outliers (*) from the 14 patients with Acute lymphoblastic leukemia (ALL) and the 19 controls (C). Unpaired Student

t test was used to compare ALL with Control group. Statistical significances are shown between groups only when p ≤ 0.05 MEIS1 Silencing Decreases the Proliferation Rate of Leukemic-derived Cell Lines Because we determined a consistent up-regulation of MEIS1 and PREP1 in cell lines and in samples A-1210477 order of patients with ALL, it was interesting to us to determine which type of advantage provides the high expression of these genes to leukemic cells. First we analyzed the role of MEIS1. The MEIS1 gene has been localized in chromosome 2 and it has been described that Jurkat cells are monosomic for this chromosome, CEM cells have two copies, and K562 cells are trisomic [21]; in this regard, expression of MEIS1 ought to be different in these cell lines. To test this hypothesis, we analyzed MEIS1 baseline

expression in these cell lines by qRT-PCR (Figure 5A). As expected, Jurkat was the cell line with the lowest MEIS1 expression, followed by CEM and K562 expressing highest levels. Taking advantage of the existing different levels of MEIS1 in the Florfenicol cell lines, we utilized Jurkat and K562 cells to investigate whether high MEIS1 expression is related with

increased proliferation. We observed that K562 have a higher proliferation rate than Jurkat cells (Figure 5B). To demonstrate the direct involvement of MEIS1 in this exacerbated proliferation, we performed silencing assays in both cell lines. We employed short hairpin RNAs shRNAs directed to two different regions of MEIS1 mRNA: one was directed to Exon 9 (E9), and the other to Exon 13 (E13). By using recombinant virus, we introduced these sequences into Jurkat and K562 cells. To assure that all infected cells were carrying the construction, a resistance gene to puromycin was also introduced and the infected cells were selected with this antibiotic. Additionally, we infected the cells with an empty virus (without shRNA) and selected them also with puromycin in order to posses a control for the selection and infection process. We then tested MEIS1 (mRNA) levels by qRT-PCR. As shown in Figures 5C and 5E, MEIS1 mRNA levels decrease with both shRNAs in both cell lines to nearly 50% of the initial expression. Employing the MEIS1-silenced cells, we then measured the proliferation rate and observed that proliferation was affected in all clones in which MEIS1 was silenced (Figures 5D and 5F).

JZ participated in the design of the study and revised manuscript

JZ participated in the design of the study and revised manuscript. JS participated in the design this website of the experiment, performed the analysis, and organized the final version of the paper. All authors read and approved the final manuscript.”
“Background Silicon nano-wires (SiNWs) have attracted the attention of many researchers due to their structural, optical, electrical and thermoelectric properties. They are expected to be

important building blocks in the future nano-electronic and photonic devices including solar cells, field-effect transistors, memory devices and chemical and biomedical sensors. Owing to their compatibility with the Si-base technology, SiNWs can be used not only as the functional units of the devices but also as the interconnects [1–6]. Various methods have been reported for SiNW fabrication, including both bottom-up and top-down techniques. Bottom-up growth methods include laser ablation, evaporation, solution-based methods and chemical vapour deposition (CVD). The CVD growth Copanlisib datasheet usually takes place via vapour-liquid-solid (VLS) route [7]. Many catalyst materials, mainly metals including Au, Al, Ni, Fe and Ag, have been used for the SiNW growth [1, 8]. Among these metals, Au as catalyst has been the most popular and most widely investigated due to its chemical inertness and low eutectic temperature of Au-Si system. However, Au introduces deep impurity levels in Si bandgap

and degrades the charge carrier mobility [8]. Therefore, alternative catalyst investigation is of crucial Thiamine-diphosphate kinase importance. One of the important parameters when considering the nano-wire fabrication process is the growth temperature, as this can determine the variety of substrates that could be used, especially when there is a prefabricated layer of some temperature-dependent material. The nano-wire growth temperature is determined by the eutectic temperature of the catalyst-precursor alloy [9]; thus,

the low-temperature growth will depend on the appropriate catalysts choice. Considering the BIBW2992 concentration characteristics of Ga, including the Ga/Si alloy low eutectic point of 29.774°C, wide temperature range for silicon solubility and its non-reactivity to form solid compound with silicon, Ga has been suggested as a good alternative to Au to grow SiNWs at low-temperatures. It is important to note that Ga does not act as catalyst for the decomposition of precursor gas as it does not assist the dissociation of SiH4 below its thermal decomposition point. Therefore, Ga acts only as a solvent, and the decomposition is achieved by plasma treatment (by the use of plasma-enhanced chemical vapour deposition (PECVD) system) [10]. In this study, Ga catalyst is used with an aim to grow SiNWs at a lowest temperature using PECVD technique. The growth temperature was varied between 100°C and 400°C. The grown nano-structures were characterised using scanning electron microscopy (SEM), Ultra Violet Visible spectroscopy (UV-Vis) and Raman spectroscopy.

Plant Ecol 149:181–193CrossRef Kessler M (2001a) Pteridophyte spe

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of plants and animals along a tropical land-use gradient. Ecol Appl 19:2142–2156PubMedCrossRef Kluge J, Kessler M, Dunn RR (2006) What drives elevational patterns of diversity? learn more A test of eFT-508 manufacturer geometric constraints, climate and species pool effects for pteridophytes on an elevational gradient in Costa Rica. selleck chemicals llc Global Ecol Biogeogr 15:358–371CrossRef Kluge J, Bach K, Kessler M (2008) Elevational distribution and zonation of tropical pteridophyte assemblages in Costa Rica. Basic Appl Ecol 9:35–43CrossRef Legendre P, Legendre L (1998) Numerical Ecology, vol 2. Elsevier, Amsterdam, pp 557–558 McCain CM (2009) Global analysis of bird elevational diversity. Global Ecol Biogeogr 18:346–360CrossRef McCune B,

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Hence, our data show that ColRS system and TtgABC pump are involv

Hence, our data show that ColRS system and TtgABC pump are involved in phenol tolerance of P. putida only under growth conditions

indicating that BAY 80-6946 ic50 especially growth-related processes of phenol tolerance are affected by both these systems. Presence of phenol in growth medium enhances proportion of cells with higher DNA content Flow cytometry is a technique which allows to analyse microbial population at single cell level and to detect distinct subpopulations with different functional and structural parameters. We have previously shown that population of solid medium-grown P. putida is heterogeneous by its DNA content and membrane permeability to propidium iodide (PI) when analysed with flow cytometry [10]. In order to assess how the wild-type P. putida and its colR- and ttgC-deficient derivatives change their population structure as well as membrane permeability when growing on different media supplemented with phenol, the microbial populations were analysed at single cell level. Flow cytometry analysis of solid medium-grown bacteria stained with the mixture of SYTO9 and PI demonstrated highly heterogeneous population structure with seven clearly distinguishable subpopulations (Fig. 4). Cells in the first three subpopulations, named as C1, BIBF1120 C2 and C3+, are considered completely

healthy as they do not stain with PI. These three populations differ from each other by their SYTO9 fluorescence intensity which most probably reflects their different DNA content. Next three populations, C1_perm, C2_perm and C3+_perm,

are considered together as cells with membrane permeable to PI but they can be also distinguished by different DNA content analogous to populations C1, C2 and C3+. This was supported by comparative analysis of SYTO9-only and SYTO9+PI-stained populations which revealed that subpopulations C1, C2 and C3+ observed with SYTO9 alone were equal to the sums of their respective healthy and PI-permeable subpopulations in case of SYTO9 and PI double tetracosactide staining (Additional File 2). Seventh subpopulation, marked as Dead, is clearly present only in glucose-grown colR-deficient cells (Fig. 4 and Additional File 3) and correlates with cell lysis. Therefore, this subpopulation most probably represents dead cells with strongly damaged membranes and even lowered DNA content. Latter is supported by observation that glucose-grown colR-deficient cells had subpopulation with remarkably lower green fluorescence when stained with SYTO9 only (Additional File 3). In addition, Dead subpopulation has lower side scatter (SSC) indicating that these cells have less complex intracellular structure compared to other cells (Additional File 3). Figure 4 Visualization of subpopulations by flow cytometry analysis. P.