oleracea and P. sativum PSII complexes (Adir 1999). Very similar results were obtained

for the N. tabacum PSII described here. If a single detergent was present in the drops, only spherulites could be grown. More promising crystals were grown in mixtures of α- or β-DDM with α- or β-OG (similar results were obtained if the n-HTG instead of OG anomers were used) (Table 2). The most successful combination contained α-DDM and β-OG. In these conditions, at least two types of morphologically distinguishable crystals were grown. The balance between the two crystal forms depended on the amount of the detergent mixture in the crystallization AUY-922 clinical trial drop (0.1–2%). With 0.2–0.5% (w/v) concentration of every component of the detergent mixture mainly group A crystals (Fig. 3) were formed after 7 days. Smaller group B crystals (Fig. 4) appeared later, after 12–15 days. An increase of the detergent concentration shifted the balance from group A to group B crystals. At the highest detergent concentrations, the growth Midostaurin purchase of group A crystals was completely suppressed and only group B crystals were formed. Fig. 3 Crystals of PSII core complex. a Typical morphology of crystals in the crystallization drops. b Diffraction pattern under cryogenic conditions with a limiting resolution of 7.0–7.8 Å. c SDS-PAGE analysis (Coomassie staining)

of the protein content of the crystals. Crystals were harvested from a crystallization drop, washed extensively and dissolved in loading buffer. Lane 1 was loaded with molecular marker, lane 2 with washing buffer and lane 3 with the solution

many containing the dissolved crystals. The complex was composed of the subunits CP47, CP43, PsbO, D1, D2 and PsbE. The subunit identification was based on the analyses of Barber et al. (1997) and Fey et al. (2008) Fig. 4 Crystals of CP43. a Typical morphology of crystals in the crystallization drops. b Diffraction pattern recorded at room temperature with a limiting resolution of 12–14 Å. c SDS-PAGE analysis of the protein content in the crystals. Lane 1 shows the molecular marker, lanes 2 and 3 (Coomassie and silver stained, respectively) show the protein sample obtained from the dissolved crystals after extensive washing. The observed single band was attributed to the CP43 subunit of PSII Analysis of group A crystals Crystals of group A could be routinely reproduced with a mixture of α-DDM and β-OG at a concentration 0.5% (w/v) and 50 mM of the H isomers of HT. Crystals grew in 6–8 days and reached a considerable size (maximal linear dimension 0.4–0.6 mm). Coomassie stained SDS-PAGE analysis of the protein mixture in the crystals showed a typical PSII core complex pattern plus the His–PsbE (Fig. 3). In order to cryoprotect crystals, a “mock” crystallization experiment without protein but with 17% PEG 400 or 22% glycerol in the usual crystallization buffer (1 mM CaCl2, 50 mM Bis–Tris, pH 7.0, 4% PEG 4000, 0.5% α-DDM, 0.

In most cases, they are single isolated forms, but they can be multiple and part of familiar syndromes such as MEN 1 syndrome, von Hippel-Lindau disease and neurofibromatosis, type 1. These are mostly (well-differentiated) tumours with relatively

slow growth, even if some of them can have an aggressive behaviour (poorly-differentiated carcinomas). The clinical picture depends on the site of the primary tumour and its ability to secrete neuroamines and peptides at supra-physiological levels (functioning tumours), able to cause a symptomatic response (clinical syndromes). Among functioning tumours, major clinical entities are: carcinoid syndrome, hypoglycaemic syndrome, Zollinger-Ellison syndrome, WDHA (Water Diarrhea-Hypo-kaliemia-Achlorydria) see more syndrome, glucagonoma syndrome. However, 90% of GEP NETs do not produce biologically active hormones (non functioning tumours) and therefore the diagnosis is often made too late, in presence of symptoms due to the mass effect and/or the presence of metastases, mainly hepatic metastases [1]. In cases at advanced stages, with a diagnostic

confirmation of metastasis, as well as in case of disease progression, the prognosis gets worse. In patients with localised well differentiated neuroendocrine carcinomas, 5-year survival is 60-100%. With regional disease or LDE225 concentration distant metastases 5-year survival is 40% and 29%, respectively [6]. As a matter of fact, the median survival in these cases is approximately 1 or 2 years. Around 80% of GEP NETs express somatostatin receptors (SSTRs), located on the cell membrane. There are five different G-protein coupled receptor subtypes (SSTRs 1-5) that are differently expressed in the various types of tumour (Table 1 and 2). Tumours expressing SSTRs often contain one or more receptor subtypes. In addition, recent studies have shown that such receptors are preferably expressed in well-differentiated forms, that some advanced tumours loose particular

Exoribonuclease receptor subtypes while keeping others [7, 8], that SSTR subtypes can form homo/heterodimers at the membrane level, to develop new receptors with different functional features [9], and that this receptor “”association”" may be induced by addition of either dopamine or somatostatin. Table 1 Somatostatin receptorsa in neuroendocrine gastroenteropancreatic tumours [%]   SSTR1 SSTR2 SSTR3 SSTR4 SSTR5 All 68 86 46 93 57 Insulinoma 33 100b 33 100 67 Gastrinoma 33 50 17 83 50 Glucagonoma 67 100 67 67 67 VIPoma 100 100 100 100 100 N-F 80 100 40 100 60 VIP, vasoactive intestinal polypeptide; N-F, Non functioning; aUsing receptor subtype antibodies; bMalignant insulinoma [Modified from Oberg K, Annals of Oncology, 2004] Table 2 Somatostatin receptor subtypes mRNA in neuroendocrine tumours.

Unstimulated or empty vaccinia stimulated cells were used as a negative control. PMA/ION stimulated cells were used a positive control. After 48 hrs of incubation, the cells were removed by washing and a biotinylated antibody against IFN-γ (10 μg/ml in PBS) was added. In the subsequent, the streptavidin conjugated with enzyme ALP was added. Finally, a precipitation substrate (BCIP) for ALP was added and the plates were incubated until spots emerged at the site of the responding cells. The spots were examined and counted in an image analyzer system. The mean number of specific spot-forming cells (SFCs) was calculated by subtracting the mean number of spots from unstimulated cells or empty vaccinia stimulated cells Bafilomycin A1 from

the mean number of spots in cells stimulated with core, E1 and E2 or core peptides or recombinant HCV poly vaccinia. Lymphocytes proliferation assay The CD4+ T cell proliferation was assessed after labeling the lymphocytes derived from the spleen

using CFSE dye (Invitrogen Molecular Probes). Labeling cells with CFSE Ten mM of CFSE stock solution was prepared by adding 90 μl Dimethyl Sulfoxide (DMSO) to 500 μg lyophilized Smoothened Agonist purchase powder of CFSE dye. The stock solution was diluted in sterile PBS/0.1% BSA to get the desired working concentration of 10 μM. Purified lymphocytes were resuspended to a concentration of 50 million cells per ml in PBS/0.1% BSA before the addition of CFSE dye. An equal volume of 10 μM of CFSE dye was added to the cell suspension in a tube 6 times more than the volume of the cell suspension and mixed well by vortexing. The labeled lymphocytes (-)-p-Bromotetramisole Oxalate were incubated for 15 min at 37°C. The staining was quenched by adding 5 volumes ice-cold complete RPMI media followed by a 5 min incubation on ice. The cells were washed three times in complete RPMI media and re-suspended in complete RPMI (2 million cells per ml for the proliferation assay and 40 million cells in 75 μl PBS for injecting to mice). To verify the CFSE-labeled cells, samples of the cell suspensions were run on a flow cytometer and were also

analyzed by fluorescent microscopy. The proliferation was assessed after stimulation of the cells with core, E1 and E2 proteins (10 μg/ml) or core peptides (10 μg/ml). PMA (10 ng/ml) and ionomycine (1 μg/ml) were added to the cells as a positive control. After adding the stimulant, the cells were incubated at 37° in 5% CO2 for 4 days. The stimulated cells were then harvested by centrifugation at 1600 rpm for 5 min. The prodedures for statining and manipulation of CFSE labeled cells should be done in the dark. Surface stain each stimulated cell with CD3 TC and CD4 PE for 3 colour flow cytometry The cells were incubated 15 min in the dark at room temperature. After washing with PBS/0.1 azide/5% FCS, the cells were immediately analyzed on FacScan or were fixed by adding an equal volume of 2% paraformaldehyde and stored overnight at 4°C before the analysis. Cells stained with CFSE have very bright fluorescence.

PubMedCrossRef 9. Noble S, Markham A. Cyclosporin. A review of the pharmacokinetic properties, clinical efficacy and tolerability MAPK inhibitor of a microemulsion-based formulation (Neoral). Drugs. 1995;50:924–41.PubMedCrossRef

10. Nashan B, Cole E, Levy G, Thervet E. Clinical validation studies of Neoral C2 monitoring: a review. Transplantation. 2002;73:S3–11.PubMedCrossRef 11. Tanaka H, Nakahata T, Ito E. Single-dose daily administration of cyclosporin A for relapsing nephrotic syndrome. Pediatr Nephrol. 2004;19:1055–8.PubMedCrossRef 12. Takeda A, Horike K, Onoda H, Ohtsuka Y, Yoshida A, Uchida K, et al. Benefits of cyclosporine absorption profiling in nephrotic syndrome: preprandial once-daily administration of cyclosporine microemulsion improves slow absorption and can standardize the absorption profile. Nephrology. 2007;12:197–204.PubMedCrossRef 13. Shirai S, Yasuda T, Tsuchida H, Kuboshima S, Konno Y, Shima Y, et al. Preprandial microemulsion cyclosporine administration is effective for patients with refractory nephrotic syndrome. Clin Exp Nephrol. 2009;13:123–9.PubMedCrossRef

14. Ehrenreich T, Churg J. Pathology of membranous nephropathy. In: Sommers SC, editor. The pathology annual no. 3. New York: Appleton-Century-Crofts; 1968. p. 145–86. 15. Cattran DC, Feehally J, Cook HT, Fervenza FC, Floege J, Gipson DS, et al. KDIGO clinical Transmembrane Transporters inhibitor practice guideline for glomerulonephritis. Kidney Int Suppl. 2012;2:S139–274. 16. Cattran DC, Alexopoulos E, Heering P, Hoyer PF, Johnston A, Meyrier A, et al. Cyclosporin in idiopathic glomerular disease associated with the nephrotic syndrome: Protein kinase N1 workshop recommendations. Kidney Int. 2007;72:1429–47.PubMedCrossRef 17. Matsuo S, Imai E, Saito T, Taguchi T, Yokoyama H, Narita I. Guidelines for the treatment

of nephrotic syndrome. Nihon Jinzo Gakkai Shi. 2011;53:78–122. 18. Rostoker G, Belghiti D, BenMaadi A, Rémy P, Lang P, Weil B, et al. Long-term cyclosporin A therapy for severe idiopathic membranous nephropathy. Nephron. 1993;63:335–41.PubMedCrossRef 19. Frische L, Budde K, Färber L, Charissé G, Kunz R, Gaedeke J, et al. Treatment of membranous glomerulopathy with cyclosporin A: how much patience is required? Nephrol Dial Transplant. 1999;14:1036–8.CrossRef 20. Iida H, Naito T, Sakai N, Aoki S. Effect of cyclosporine therapy on idiopathic membranous nephropathy presented with refractory nephrotic syndrome. Clin Exp Nephrol. 2000;4:81–5.CrossRef 21. Rifai N, Chao FF, Pham Q, Thiessen J, Soldin SJ. The role of lipoproteins in the transport and uptake of cyclosporine and dihydro-tacrolimus into HepG2 and JURKAT cell lines. Clin Biochem. 1996;29:149–55.PubMedCrossRef 22. Sugioka N, Kokuhu T, Okamoto M, Yoshimura N, Ito Y, Shibata N, et al. Effect of plasma lipid on pharmacokinetics of ciclosporin and its relationship with plasma prednisolone level in renal transplant patients. J Pharm Pharmacol. 2006;58:1193–200.PubMedCrossRef 23. Brunet M, Campistol JM, Millán O, Vidal E, Esforzado N, Rojo I, et al.

Moreover, thermal quenching is found to be more severe for the high energy PL components which lead to an apparent red shift of the PL maximum position at high T. To get further insights into the mechanisms responsible for the observed thermal quenching, we have analyzed Arrhenius plots of the PL intensity at

different detection energies (E det) as shown in Figure  2a. The analysis was performed for constant detection JQ1 in vivo energies since (a) the temperature-induced shift of the bandgap energy is significantly suppressed in GaNP alloys [15], and (b) spectral positions of the excitons bound to various deep-level N-related centers do not one-to-one follow the temperature-induced shift of the bandgap energy. This approximation defines error bars of the deduced values as specified below. All experimental data (shown by the symbols in Figure  2) can be fitted bywhere I(T) is the temperature-dependent PL intensity, I(0) is its value at 4 K, E 1 and E 2 are the activation energies

for two different thermal quenching processes, and k is the Boltzman constant (the results of the fitting are shown by the solid lines in Figure  2a). The first activation process that occurs within the 30 to 100 K temperature range is characterized by the activation energy E 1 ranging between 40 (at E det = 2.17 eV) and 60 meV (at E det = 2.06 eV). The contribution of this process is most pronounced for high energy PL components that correspond to the radiative recombination at the N-related localized states with CT99021 price their energy levels close to the GaNP band

edge. The quenching of the high energy PL components is accompanied by a slight increase in the PL intensity at low E det. Therefore, this process can be attributed to the thermal ionization of the N-related localized states. Such ionization is expected to start from the N-states that are shallower in energy. The thermally activated excitons can then be recaptured by the deeper N states, consistent with our experimental observations. We note that the determined values of E 1 do not one-to-one correspond to the ‘apparent’ depth of the involved localized states deduced simply from the distance between E det and the bandgap energy of the GaNP. Celastrol This is, however, not surprising since such correspondence is only expected for the no-phonon excitonic transitions whereas recombination of excitons at strongly localized states (such as the monitored N states) is usually dominated by phonon-assisted transitions due to strong coupling with phonons. Figure 2 Arrhenius plots of the PL intensity measured at different detection energies from the GaP/GaNP NWs (a) and GaNP epilayer (b). (1) The second thermal quenching process is characterized by the activation energy E 2 of approximately 180 ± 20 meV, which is the same for all detection energies. This process becomes dominant at T > 100 K and leads to an overall quenching of the PL intensity irrespective of detection energies.

A hyphen indicates that the branch was not obtained with the respective reconstruction method. Nucleotide sequence accession numbers are given in parentheses. The affiliation of strains to subclades of the OM60/NOR5 group is based on [13]. The sequence of Alcanivorax borkumensis [GenBank:Y12579] was used as outgroup (not shown). Designations given in red color indicate that the respective strains produce BChl a and/or encode genes for a photosynthetic apparatus; names in blue indicate the presence of proteorhodopsin encoding genes. Strains that were tested with specific PCR primers for the presence of pufLM and soxB genes are labeled with red and yellow circles,

respectively. Closed circles indicate a positive PCR reaction and open circles a negative reaction. The bar represents an estimated sequence divergence of 5%. It was not possible to amplify genes encoding proteorhodopsin Panobinostat datasheet or the sulfate thiol esterase SoxB from the non-phototrophic species shown in Figure  1. For the PCR screening with

the proteorhodopsin primer set PR1-3 [26] we used genomic DNA from Dokdonia sp. PRO95 [27] as well as total DNA isolated from the North Sea as positive control. However, a proteorhodopsin-positive control strain belonging to this phylogenetic group was not available and the pop gene sequence of strain IMCC3088 revealed some mismatches to the used proteorhodopsin oligonucleotide primers. Thus, either the tested strains do not encode pop genes, or the genes are such different at the primer binding sites that no PCR amplification was possible. Phenotypic characterization Morphology ICG-001 of cells and colonies Size and shape of cells of the newly isolated Docetaxel strain Ivo14T were determined upon growth in SYPHC medium, which was optimal for cultivation of this strain and the related species C. litoralis, H. rubra and Chromatocurvus halotolerans. Cells of Ivo14T were non motile and appeared

coccoid or as short straight-to-bent rods. Occurrence of pleomorphic cells was observed in all four BChl a-containing strains and depended to some extent on the composition of the growth medium, which makes it important to use the same medium for comparison of size and shape. Especially, growth on the nutrient-rich medium Marine Broth 2216 led in cultures of H. rubra, C. litoralis and Chromatocurvus halotolerans to cells with irregular shapes, swelling of cells and accumulation of highly refractile storage compounds, whereas these effects were less pronounced in cultures of Ivo14T. The storage compound cyanophycin, which is a characteristic of C. litoralis was not detected in cells of Ivo14T or Chromatocurvus halotolerans, which both accumulate polyhydroxyalkanoates in addition to polyphosphates. The intracellular carbon storage compound of H. rubra could be distinguished from cyanophycin or polyhydroxyalkanoates by a positive reaction of the acidified cell extract with the anthrone reagent, which detects carbohydrates.

2009; Proteasome inhibitor Zhang et al. 2009a). Concluding remarks The familial status of Neophaeosphaeria under Leptosphaeriaceae is confirmed, although this family remains poorly supported in phylogenetic studies. Nodulosphaeria Rabenh., Klotzschii Herb. Viv. Mycol., Edn 2: no. 725 (in sched.) (1858). (Phaeosphaeriaceae) Generic description Habitat terrestrial, saprobic or

hemibiotrophic. Ascomata small, immersed to erumpent, globose or subglobose, black, papillate, ostiolate. Papilla with numerous setae in the pore-like ostiole. Peridium thin, composed of thick- or thin-walled large cells. Hamathecium of cellular pseudoparaphyses, septate and branching. Asci 8-spored, bitunicate, fissitunicate, clavate to cylindro-clavate, with a very short, furcate pedicel and a small ocular chamber. Ascospores filamentous, hyaline or pale brown, multi-septate, one of the upper cells swollen. Anamorphs reported for genus:

none. Literature: Barr 1992a; Holm 1957, 1961; Shoemaker 1984b; Shoemaker and Babcock 1987. Type species Nodulosphaeria hirta Rabenh., Klotzschii Herb. Viv. Mycol., Edn 2: no. 725 (in sched.) (1858). (Fig. 67) Fig. 67 Nodulosphaeria hirta (from BR 101945–95, holotype). a Appearance of ascomata on the host surface. b Vertical section of an ascoma. Note the setae at the apex and in the ostiole. c Section of a partial peridium. Note the outer layer cells of textura angularis and inner layer compressed cells. d Squash mount showing asci in pseudoparaphyses. e, f. The light brown filiform ascospores. Scale bars: a = 0.5 mm, b = 100 μm, c = 50 μm, find more d = 20 μm, e, f = 10 μm Ascomata 260–330 μm high × 260–330 μm diam., scattered, or in small groups, immersed to erumpent, globose or subglobose, black, papillate, ostiolate. Papilla 50–80 μm high, numerous setae occur in the pore-like ostiole (Fig. 67a and b). Peridium 15–30 μm wide at the sides, thinner at the base, coriaceous, comprising two types of cells, outer cells of 1–2 layers of heavily pigmented cells of textura angularis, cells 6–8 μm diam., cell wall Methane monooxygenase 1.5–3 μm thick, inner of compressed cells,

5 × 13–3 × 8 μm diam., wall 2–3 μm thick (Fig. 67c). Hamathecium of long cellular pseudoparaphyses 2–3 μm broad, septate and branching, mucilage not observed. Asci 100–123 × 12.5–15(−17.5) μm (\( \barx = 110.8 \times 14.3\mu m \), n = 10), 8-spored, bitunicate, fissitunicate, clavate to cylindro-clavate, with a very short, furcate pedicel, with a small ocular chamber (to 2 μm wide × 1 μm high) (Fig. 67d). Ascospores 48–63 × 5–6.5 μm (\( \barx = 55.3 \times 5.6\mu m \), n = 10), 4-seriate, filamentous, pale brown, 8-septate, the 4th upper cell broader than the others, smooth-walled, without sheath (Fig. 67e and f). Anamorph: none reported. Material examined: GERMANY, Dresdae, in herbarum caulibus emortuis perrara, exeunte majo, 1858 (BR 101945–95, holotype, as Nodulosphaeria hirta).

17,048 WGHs are found in the 1,668 eukaryotic genomes. The top three phyla in the numbers of FACs are also top three in the numbers of WGHs; and 2,328, 5,444 and 5,171 WGHs are encoded in three phyla Arthropoda, Ascomycota and Streptophyta, respectively. The top four eukaryotic genomes in the numbers

of WGHs are from the phylum Streptophyta, and they are Oryza sativa sp japonica (Rice) (828 WGHs), Arabidopsis thaliana (Mouse-ear cress) (678 WGHs), Vitis vinifera (Grape) (602 WGHs) and Zea mays (Maize) (284 WGHs). It is interesting to observe that there are 272 and 224 WGHs in the human and mouse genomes, respectively. Besides two other plant genomes, i.e. Oryza sativa subsp. indica (Rice) (258 WGHs) and Physcomitrella patens

PD0325901 cell line sp patens (Moss) (226 WGHs), all the other 6 eukaryotic genomes encoding more than 200 WGHs are from the fungal phylum Ascomycota. No cellulosome components were identified in the eukaryotic genomes. 200 learn more (~73.53%) human WGHs are homologous to mouse WGHs with NCBI BLAST E-values < e-23. So the majority of these enzymes have been in the genomes of human and mouse at least before their divergence 75 million years ago [36]. Identified glydromes in metagenomes Overall, 63 FACs and 6,072 WGHs are found in 42 metagenomes except for TM7b which was sampled from the human mouth. The top two metagenomes in the numbers of glycosyl hydrolases are from termite guts (12 FACs and 1,150 WGHs) and diversa silage soil (13 FACs and 820 WGHs). Since the number of proteins in metagenomes varies from 452 in termite gut fosmids to 185,274 in the diversa silage soil, we calculated the percentage of the glycosyl hydrolases in each metagenome. On average, 0.65% of a metagenome encode glycosyl hydrolases. We noted that all the metagenomes with

more than 1% encoding glycosyl hydrolases are from the animal guts (including Progesterone human, mouse and termite). This is confirmed by an independent study using BLAST mapping [37]. No cellulosome components were identified in any metagenome. Utility The query interface of GASdb All the annotated glydromes were organized into an easy-to-use database GASdb (Figure 2). A user can find the proteins of interest through browsing, and searching using keywords or BLAST. The overall organization of each glydrome can be displayed; and the high resolution images of each protein can be downloaded for the publication purpose, as shown in Figure 3. A user can also display the signal peptide and functional domains of a given protein and its homologs using BLAST with E-value cutoff 1e-20, as shown in Figure 3. Figure 2 The database interfaces: the main page, the browsing page, the searching page, and the BLAST page. Figure 3 The displaying pages for the domain architectures of the glydrome of Clostridium acetobutylicum , and domain architectures of the protein Clostridium acetobutylicum CelA and its homolog.

25) and for all further analysis the wave velocities of both strains were combined. Availability of supporting data The data sets supporting the results of this article are available in the 3TU.Datacentrum repository [56], [doi:10.4121/uuid:f5603abf-bf15-4732-84c0-a413ce7d12d3], [http://dx.doi.org/10.4121/uuid:f5603abf-bf15-4732-84c0-a413ce7d12d3]. Acknowledgments We thank Martin Ackermann, Robert H. Austin, Jean-Baptiste

Boulé, Cees Dekker, Alex Hall, Rutger Hermsen and Pieter Schoustra for valuable comments and discussion CYC202 in vitro and Orsolya Haja for measuring the bulk growth curves. The project described was supported by Grant Number U54CA143803 from the National Cancer Institute. The content is solely the responsibility of the authors and does

not necessarily represent the official views of the National Cancer Institute or the National Institutes of Health. P.G. was supported by the “Lendület” program of the Hungarian Academy of Sciences. Electronic supplementary material Additional file 1: Growth curves of strains JEK1036 and JEK1037 in bulk conditions. Growth curves are shown for strains JEK1036 (in green) and JEK1037 (in red), for each strain 3 independent cultures were grown in 200 ml LB in 500 ml flasks at 30°C. For each sample the OD600 was measured in triplicate and their average value was check details used. Error bars indicate sem. The inset shows the growth curve using linear y-scale for the first 15 hours. (PDF 104 KB) Additional file 2: Overview of all devices with separate inlets (type 1).

(A) Each kymograph shows the average occupancy per patch in a single habitat. Kymographs for the five parallel habitats in a single device are shown next to each other. Note that all habitats on the same device are inoculated from the same culture set. (B) The device-wide averages of the occupancies of strains JEK1037 (R red) and JEK1036 (G green) and the red fraction (f r black) are shown as function of time. Dashed lines indicate mean ± sem. The red fraction (f r ) is calculated for each habitat as f r  = r/(r + g), where r and g are the habitat-wide average G protein-coupled receptor kinase occupancies of strains JEK1037 (red) and JEK1036 (green) respectively. Habitats where one (or both) of the strains failed to enter (e.g. when there is a constriction in one of the inlet channels) were excluded from the analysis and are shown as grey panels in this figure. (PDF 443 KB) Additional file 3: Overview of all devices with a single inlet (type 2). (A) Each kymograph shows the average occupancy per patch in a single habitat. Kymographs for the five parallel habitats in a single device are shown next to each other. Note that all habitats on the same device are inoculated from the same culture set. (B) The device-wide averages of the occupancies of strains JEK1037 (R, red) and JEK1036 (G, green) and the red fraction (f r black) are shown as function of time. Dashed lines indicate mean ± sem.

We investigated

the morphology and structure of the as-obtained precipitate by TEM, SEM, and SAED, respectively. When the solvent of the whole system is only water (none of EG), a dark-green precipitate is produced immediately after the FeSO4 solution is dropped into excessive NaOH solution. In contrast to pure aqueous solution, the precipitate of ferrous hydroxide in the H2O-EG mixture solution was white at the beginning and turns green then dark-green gradually. The precipitate of ferrous hydroxide obtained in pure aqueous solution is also known as ‘green rust’ in the crystal lattice of which iron(II) ions are easily substituted by iron(III) ions produced by its progressive oxidation [35–37]. However, the oxidation process is inhibited in the H2O-EG mixture solution because of the reducing power of EG. All forms of green rust 5-Fluoracil are more complex and variable than the ideal iron(II) hydroxide compound. TEM images of the precipitate (Figure 4a) obtained in H 89 concentration pure aqueous solution show that there are two kinds of products at least; one of them is a very thin nanoplate with a diameter of about 50 nm, and the other is a needle-shaped nanoparticle. TEM and SEM images (Figures 4b and 5a,b) of the end product of this precipitate after aging for 24h in 90°C show that the obtained product is a mixture of polygonal particles and fiber-like particles. The sizes

of the polygonal particles are about 50 to 100 nm. However, no rod-like or fiber-like nanoparticles can be found in the TEM and SEM images of the as-obtained ferrous hydroxide precipitate (Figure 4c,d) in the H2O-EG mixture solution. Ferrous hydroxide obtained in the H2O-EG mixture solution forms a large-scaled film rather than plate-like and rod-like nanoparticles in pure aqueous solution. Also, according to its SAED pattern (Figure 4e), the ferrous hydroxide film has a polycrystalline structure. TEM and SEM images of the Fe3O4 nanoplate obtained in the EG-H2O mixture solution with the ratio of EG/H2O = 3:1 and 5:1 are shown in Figure 5c,d,e,f. It

can be seen that the thickness of the Fe3O4 nanoplates decreases, and the shape of the nanoplate becomes more irregular when the concentration of EG increases. From the analysis of the above experiments, Ribonucleotide reductase it is obvious that the addition of EG affects the formation of Fe3O4 nanoplate. Figure 4 Fe(OH) 2 and the as-prepared Fe 3 O 4 . (a) TEM images of Fe(OH)2and (b) low-magnification SEM images of the as-prepared Fe3O4obtained in pure aqueous solution. It can be seen that the product is a mixture of polygonal particles and fiber-like particles. (c) SEM and (d) TEM images and (e) the SAED pattern of Fe(OH)2 obtained in the EG-H2O mixture. Figure 5 The Fe 3 O 4 nanoparticles and nanoplates prepared under different conditions. (a) TEM and (b) SEM images of the as-prepared Fe3O4 nanoparticle (EG/H2O = 0:1). (c) TEM and (d) SEM images of Fe3O4 nanoplates prepared under the condition of EG/H2O = 3:1.