It has also been suggested that the two components of this particular regulatory system do not always act in tandem specifically in response to acid stress. From the results obtained in this study, we cannot speculate on the overexpression of CpxA in PA adapted cultures-as CpxA is a membrane localized protein and this study focused on soluble proteins. It may be informative, however, to examine the expression profile of CpxA in PA adapted cultures in order to decipher if CpxR works in a concerted manner with CpxA to protect cells from acid stress following the onset of PA-induced acid resistance. Conclusion

It is apparent that long ACP-196 nmr term PA adaptation of S. Enteritidis is associated with differential protein expression, with the synthesis of

certain proteins being significantly upregulated. selleck screening library Of these proteins, Dps and CpxR are those commonly associated with virulence and we have not only demonstrated that they are inducible by PA, but also that they are crucial for PA-induced acid resistance in S. Enteritidis. These results clearly demonstrate that Dps and CpxR play an important role in PA-induced acid resistance. It is also apparent that overexpression of either Dps or CpxR alone in PA adapted cultures is not sufficient to confer increased acid resistance. Acknowledgements This study was supported by a USDA Food Safety Consortium grant. Electronic supplementary material

Additional file 1: Protein https://www.selleckchem.com/products/MS-275.html report C. Mass spectrometry report for RplE (PDF 370 KB) Additional file 2: Protein Report B. Mass spectrometry report for RplF (PDF 262 KB) Additional file 3: Protein Report A. Mass spectrometry report for SodA (PDF 343 KB) Additional file 4: Protein Report D. Mass spectrometry report for CpxR and Dps (PDF 345 KB) References 1. Callaway TR, Edrington TS, Anderson RC, Byrd JA, Nisbet DJ: Gastrointestinal microbial ecology and the safety of our food supply as related to Salmonella . J Anim Sci 2008,86(E suppl):E163-E172.PubMed 2. Foster JW, Hall HK: Adaptive Acidification Thiamine-diphosphate kinase Tolerance Response of Salmonella typhimurium . J Bacteriol 1990, 172:771–778.PubMed 3. Lee IS, Slonczewski JL, Foster JW: A Low-pH-Inducible, Stationary-Phase Acid Tolerance Response in Salmonella typhimurium . J Bacteriol 1994, 176:1422–1426.PubMed 4. Lin J, Lee IS, Frey J, Slonczewski JL, Foster JW: Comparative Analysis of Extreme Acid Survival in Salmonella typhimurium , Shigella flexneri , and Escherichia coli . J Bacteriol 1995, 177:4097–4104.PubMed 5. Kwon YM, Ricke SC: Induction of acid resistance of Salmonella typhimurium by exposure to short-chain fatty acids. Appl Environ Microbiol 1998, 64:3458–3463.PubMed 6. Gahan CG, Hill C: The relationship between acid stress response and virulence in Salmonella typhimurium and Listeria monocytogenes . Int J Food Microbiol 1999, 50:90–100.CrossRef 7.

Western blotting Preparation of nuclear extracts for NF-κB 4T1 and NMuMG cells treated under various conditions were washed with cold PBS and suspended buy SIS3 for 30 min in 0.4 ml of a hypotonic lysis buffer (20 mM Tris–HCl (pH 7.5), 10 mM NaCl, 1 mM EDTA, 2 mM Na3VO4,) containing protease inhibitors (10 μg/ml leupepton, 1 μM pepstatin). The cells were then lysed with 12.5 μl of 10% nonyl phenoxylpolyethoxylethanol (NP-40). The homogenate was centrifuged, and the supernatant, which contained the

cytoplasmic extracts, was stored at −80°C. The nuclear pellet was resuspended in 25 μl of ice-cold nuclear-extraction buffer for 30 min, with intermittent mixing. Then, the extract was centrifuged, and the supernatant containing the nuclear extract was obtained.

The protein content was measured by using the BCA protein assay kit (Pierce, Rockford, IL, USA). The nuclear and cytoplasmic extracts (40 μg of protein) were fractionated on polyacrylamide-sodium dodecyl sulfate (SDS) gels and transferred to polyvinylidene fluoride (PVDF) membranes (Amersham, Selleck DZNeP Arlington Heights, IL, USA). The membranes were blocked with a solution containing 3% skim milk and incubated with the anti-NF-κB p65 antibody (Cell Signaling Technology, Beverly, MA, USA) overnight at 4°C. Subsequently, the membranes were incubated with anti-rabbit IgG sheep antibody coupled to horseradish peroxidase (Amersham) for 1 h at room temperature. The reactive proteins were visualized by using ECL-plus (Amersham) according to the Selleck PU-H71 manufacturer’s instructions. Anti-lamin A antibody (Santa Cruz Biotechnologies, CA, USA) was used as the internal standard; it was used as the primary antibody to detect lamin Progesterone A. Preparation of whole-cell lysates 4T1 and NMuMG cells treated

under various conditions were lysed with a lysis buffer containing 20 mM Tris–HCl (pH 8.0), 150 mM NaCl, 2 mM EDTA, 100 mM NaF, 1% NP-40, 1 μg/ml leupeptin, 1 μg/ml antipain, and 1 mM phenylmethylsulphonyl fluoride. The protein content in the cell lysates was determined using a BCA protein-assay kit. The extracts (40 μg of protein) were fractionated on polyacrylamide-SDS gels and transferred to PVDF membranes (Amersham). The membranes were blocked with a solution containing 3% skim milk and incubated overnight at 4°C with each of the following antibodies: anti-NF-κB p65, anti-phospho-extracellular signal-regulated kinase (ERK) 1/2 antibody, anti-phospho-Akt antibody, anti-phospho-mammalian target of rapamycin (mTOR) antibody, anti-phospho-c-Jun N-terminal kinase (JNK) antibody, anti-phospho-signal transducers and activator of transcription 3 (STAT3) antibody, anti-ERK1/2 antibody, anti-Akt antibody, anti-mTOR antibody, anti-JNK antibody, and anti-STAT3 antibody (Cell Signaling Technology).

Alignments learn more of multiple protein sequences to view areas of conservation amongst A domains were performed using Clustal W http://​www.​ebi.​ac.​uk/​ Generation of 3D-models for FnBPB (N23) types I-VII Theoretical models of the structure of

region A (N23) types I-VII were obtained by submitting the amino acid sequences for this segment of each protein to the Phyre service of the 3D-PSSM website http://​www.​sbg.​bio.​ic.​ac.​uk/​phyre/​. This web-based tool models the structure of these sequences based structure of the equivalent domains of the S. aureus clumping factor ClfA. All structures were viewed using the pyMOL viewing software. Expression of recombinant FnBPB A domain proteins Primers were designed to amplify DNA encoding residues Palbociclib ic50 162-480 (N23 sub-domain) of FnBPB isotype I from strain 8325-4 by PCR. The primers included BamHI and SmaI restriction sites to facilitate cloning into the multiple cloning site of the N-terminal six-histidine tag expression

vector pQE30 (Qiagen) and incorporated a 3′ stop codon. The equivalent N23 regions of FnBPB isotypes types II-VII were PCR-amplified from strains N315, MSSA476, P1, 2, 3077 and 233, respectively. The PCR products were cloned separately into pQE30 and transformed into E. coli cells for protein production. Each construct was verified by sequencing (GATC Biotech AG, Germany) and proteins were purified by selleck chemicals Ni2+ chelate chromatography [35]. Concentrations were determined using the BCA Protein Assay Kit (Pierce). Proteins were dialysed against PBS for 24 h at 4°C, aliquoted and stored at -70°C. Direct binding of recombinant FnBPB A domain proteins to immobilized elastin, fibrinogen

and fibronectin Human aortic elastin (Elastin Products Company; 50 μg/ml) was coated onto microtiter wells for 18 hr under UV light. Wells coated with human fibrinogen (Calbiochem; 10 μg/ml), and fibronectin (Calbiochem; 10 μg/ml) were placed at 4°C overnight. All plates were blocked with 5% skimmed milk in phosphate http://www.selleck.co.jp/products/cobimetinib-gdc-0973-rg7420.html buffered saline (PBS) for 2 hr at 37°C. Following three washes with PBS containing 0.05% v/v Tween 20 (PBST) various concentrations of purified rFnBPB N23 constructs in PBS were added and incubated at 37°C for 2 hr. After three washes with PBST, bound protein was detected by incubation with a 1:500 dilution of monoclonal antibody 7E8 that recognizes the N-terminal hexahistidine fusion tag. After 1 h incubation with shaking at room temperature, the wells were washed three times with PBST followed by 100 μl per well of goat-anti-mouse IgG antibodies conjugated to horseradish-peroxidase (HRP, Dako; Denmark) diluted 1:2000. After incubation for 1 h at room temperature, wells were washed three times with PBST, and bound HRP-conjugated antibodies were detected with 10 μg per well of 3,3′,5,5′-tetramethylbenzidine (TMB; Sigma) in 0.05 M phosphate-citrate buffer containing 0.006% (v/v) hydrogen peroxide.

Evidence also suggests that glucocorticoids may inhibit the action of leptin [27]. Results from a number of studies indicate a general endocrine response to hypocaloric diets that promotes increased hunger, reduces metabolic rate, and threatens the maintenance of lean mass. Studies involving energy restriction, or very low adiposity, report decreases in leptin [1, 10, 28], insulin [1, 2], testosterone [1, 2, 28], and thyroid hormones [1, 29]. Subsequently, increases in ghrelin [1, 10] and cortisol [1, 30, 31] have

been reported with energy restriction. Further, there is evidence to suggest that unfavorable changes in circulating hormone levels persist as subjects attempt to maintain a reduced body weight, even after the cessation of active weight loss [32, 33]. 4SC-202 concentration Low energy intake and minimal body fat are perceived HM781-36B as indicators of energy unavailability, resulting in a homeostatic endocrine response aimed at conserving energy and promoting energy intake. It should be noted that despite alterations in plasma levels of anabolic and catabolic hormones, losses of lean body mass (LBM) often fail to reach statistical significance in studies on bodybuilding

preparation [1, 2]. Although the lack of significance may relate to insufficient statistical power, these findings may indicate that unfavorable, hormone-mediated changes in LBM can potentially be attenuated

by sound training and nutritional practices. Previous research has indicated that structured resistance training [34] and sufficient protein intake [35–37], both commonly employed in bodybuilding contest preparation, preserve LBM during energy restriction. Further, Maestu et al. speculate that losses in LBM are dependent on the magnitude of weight loss and degree of adiposity, as the subjects who lost the greatest amount of weight and achieved the lowest final body fat percentage in the study saw the greatest losses of LBM [2]. The hormonal environment created by low adiposity and energy AICAR cell line restriction appears to promote weight regain and threaten selleck chemicals llc lean mass retention, but more research is needed to determine the chronic impact of these observed alterations in circulating anabolic and catabolic hormones. Weight loss and metabolic rate An individual’s total daily energy expenditure (TDEE) is comprised of a number of distinct components (Figure 1). The largest component, resting energy expenditure (REE), refers to the basal metabolic rate (BMR) [8]. The other component, known as non-resting energy expenditure (NREE), can be further divided into exercise activity thermogenesis (EAT), non-exercise activity thermogenesis (NEAT), and the thermic effect of food (TEF) [8]. Figure 1 Components of total daily energy expenditure (TDEE).

J Am Geriatr Soc 2001,49(12):1691–1699.PubMedCrossRef 17. Min LC, Elliott MN, Wenger NS, Saliba D: Higher vulnerable elders survey scores predict death and functional decline in vulnerable older people. J Am Geriatr Soc 2006,54(3):507–511.PubMedCrossRef 18. Min L,

Yoon W, Mariano J, Wenger NS, Elliott MN, Kamberg C, et al.: The vulnerable elders-13 survey predicts 5-year functional decline and mortality outcomes in older ambulatory care patients. J Am Geriatr Soc 2009,57(11):2070–2076.PubMedCrossRef 19. EuroQol group. Health policy: A new facility ATM inhibitor for the measurement of health-related quality of life. Health Policy 1990,16(3):199–208.CrossRef 20. Deiner S, Silverstein J: Long term outcomes in elderly surgical patients. Mt Sinai J Med

2012,79(1):95–106.PubMedCrossRef Competing interests The authors declare that they have no competing interests. Authors’ contributions SG: Study design, data analysis, data interpretation, Tideglusib and writing, YT: data collection, writing, AW: study design, data interpretation, and critical revision, SLW: Study design, data interpretation, and critical revision, RGK: Study design, data analysis, data interpretation, and critical revision. All authors read and approved the final manuscript.”
“Background War is a type of collective violence which is defined as an instrumental use of violence by members of a group against another in order to achieve political, economic or social objectives [1]. The highest rates of war-related deaths are in the WHO African Region followed by parts of the WHO Eastern Mediterranean region. More than half a million people died during the first Gulf War (1980–1988) between Iraq and Iran [1]. Explosive weapons are designed to increase the number and energy of casing fragments leading to multiple penetrating wounds [2]. This is why vascular injuries are often associated with multiple trauma leading to high mortality unless prompt and appropriate surgical management is made. The evacuation time, climate, and availability of medical resources

will impact the outcome of surgical management of war-injured patients [3]. Shortening aminophylline the evacuation time in the prehospital setting reduced the war-related mortality [4–6], while prolonged evacuation resulted in high mortality [7]. Ideally, war injuries should be treated by surgeons having military surgery experience. In fact, civilian surgeons may find themselves trapped in wars practicing military surgery without prior training or experience in this field [4]. The mechanism and pattern of vascular injury will vary in the same community in war and peace. The commonest mechanism of injury in civilian practice in most parts of the world is road traffic collisions. We have found in a prospective cohort study that vascular injuries constituted 1.2% of all hospitalized motor vehicle Selonsertib ic50 collision trauma patients in a civilian setting [8].

After permeabilization with 0.1% Triton X-100 (in 1X PBS) for 10 min at room temperature, cells were incubated with 0.1 M Glycine (in 1X PBS) and attached to glass coverslips coated with 0.1% poly-L-Lysine (Sigma). Anti-LaTRF serum was used selleck chemicals to detect LaTRF with Alexa Fluor 555-labeled goat anti-rabbit IgG (Invitrogen) as the secondary

antibody followed by telomere detection using a Telomere PNA FISH Kit/FITC (DakoCytomation). VECTASHIELD® Mounting Medium with DAPI (Vector Labs) was used as the anti-fade mounting solution and to stain nuclear and kinetoplast DNA. The images were analyzed with a Nikon 80i fluorescence microscope and captured with a digital camera (Nikon). When necessary, images were superimposed using NIS elements software (v. Br 2.30). EMSA (electrophoretic mobility shift assay) All of the conditions for binding reactions and EMSA, including binding temperature, PD173074 protein concentrations in the extracts and the double-stranded DNA probe (LaTEL), were standardized in preliminary experiments. LaTEL was constructed by using the γ [32P]ATP 5′-end-labeled oligonucleotides ssTel78G and ssTel78C, as described by Lira et al. [17]. Assays were done by mixing 10 μg of renatured bacterial extracts containing full length LaTRF or LaTRF Myb

with approximately 2 pmol of labeled probe (LaTEL) in 30 μl of EMSA buffer (20 mM HEPES, 2.5 Alvocidib ic50 mM MgCl2, 0.1 mM EDTA, 0.1 M KCl, 10% glycerol, 0.5 mM DTT, pH 8.0) containing 10 ng of poly [dI-dC] [dI-dC] and 10 ng of poly [dA-dT] [dA-dT]. Total protein extracts of non-transformed E. coli were used as controls. The reactions were incubated for 30 min at room temperature and loaded onto a non-denaturing 4% polyacrylamide gel (acrylamide:bis-acrylamide, 19:1, w/w) in 1X TBE. After electrophoresis, the gels were exposed to X-ray film. Binding reactions were also done with crude nuclear extracts obtained from 108 parasites

(~2.3 μg of total proteins) and pheromone γ [32P]ATP labeled LaTEL (2 pmol) in EMSA buffer containing a mixture of 10 ng of poly [dI-dC] [dI-dC] and 10 ng of poly [dA-dT] [dA-dT]. Competition assays to test the binding specificity of proteins in both recombinant and nuclear extracts, were done using 20 fold excess of unlabeled LaTEL (in relation to the labeled probe) as the specific competitor and a 100 fold excess (in relation to the labeled probe) of unlabeled double-stranded DNA poly [dI-dC] [dI-dC] as the non-specific competitor. Supershift assays were done using full-length recombinant LaTRF (10 μg) or native nuclear extracts from 108 parasites in the presence of ~30 μg of anti-LaTRF serum in EMSA buffer containing labeled LaTEL as probe and both poly [dI-dC] [dI-dC] and poly [dA-dT] [dA-dT] as above described. These assays were also performed in the presence of 20 fold excess of non-labeled LaTEL and 100 fold excess of poly [dI-dC] [dI-dC] as described above. Chromatin immunoprecipitation Formaldehyde cross-linked chromatin was obtained from promastigote forms of L.

The foreseen ways of the further Al-BNNT composite enhancement are proposed by us as follows: (1) increasing the BNNT loading fraction and the tube texturing/alignment in a given matrix, (2) functionalization and/or perforation of the external BNNT surfaces to increase their cohesion with the Al matrix, (3) pre-heat treatment of the ribbons before the tensile tests directed to the second

phase precipitation at the BNNT-Al interfaces and increasing the efficiency of a load transfer via this website chemical bonding at the nanotube-metal interfaces, and (4) trying advanced powder metallurgy routes, i.e., spark-plasma sintering, to fabricate ultimately denser and larger BNNT-containing lightweight Al-based composites. Finally, it could be mentioned that combination of BNNTs and BN nanosheets [7] as a reinforcing phase in Al-based composites may also be an interesting direction. Such complex hybrids may possess an enhanced efficiency of the load transfer from a weak Al matrix to the strong and resilient

nano-BN phases. These are the topics of our ongoing research. Conclusions In summary, for the first time, we fabricated Al-BNNT composite ribbons (up to 1 m long) with various multiwalled BNNT contents (0.5 to 3.0 wt.%) by melt spinning. Scanning and transmission electron microscopy, X-ray diffraction, and energy selleck chemicals llc dispersive X-ray analysis confirmed the decent integration of the two phases into a dense and compact composite. No other phases, like Al borides or nitrides, AS1842856 form in the resultant melt-spun composites. The BNNTs are randomly oriented within the Al matrix and partially participate in carrying the tensile load, as evidenced by their presence and breakage at the composite fracture surfaces. The ultimate tensile strength of the composite ribbons with 3 wt.% of BNNT at room temperature was more than doubled (145 MPa) compared to non-loaded

pure Al ribbons (60 MPa). Acknowledgements This work was supported by the World Premier International (WPI) Center for Materials Nanoarchitectonics (MANA) tenable at the National Institute for Materials Science (NIMS), Tsukuba, Japan. D.G. also acknowledges a funding ‘Mega-Grant’ award for leading scientists tenable Benzatropine at the National University of Science and Technology “MISIS”, Moscow, Russian Federation under the agreement no. 11.G34.31.0061. The authors thank Prof. K. Hono for his permission for using a melt-spinning machine and Drs. P. Delhibabu, S. A. Hossein, M. Mitome, and N. Kawamoto of MANA-NIMS for their technical support. M.Y. and D.G. particularly acknowledge a financial support from a grant-in-aid no. 23310082 (‘Kakenhi’, Japan Society for Promotion of Science, JSPS). References 1. Bakshi SR, Lahiri D, Agarwal A: Carbon nanotube reinforced metal matrix composites – a review. Inter Mater Rev 2010, 55:41–64.CrossRef 2.

J Biol Chem 2001,

276 (52) : 48899–48907.CrossRefPubMed 25. Kubo E, Urakami T, Fatma N, Akagi Y, Singh DP: Polyol pathway-dependent osmotic and oxidative stresses in aldose reductase-mediated apoptosis in human lens epithelial cells: role of AOP2. Biochem Biophys Res Commun 2004, 314 (4) : 1050–1056.CrossRefPubMed 26. Váli L, Hahn O, Kupcsulik P, Drahos A, Sárváry E, Szentmihályi K, Pallai Z, Selisistat Kurucz T, Sípos P, Blázovics A: Oxidative stress with altered element content and decreased ATP level of erythrocytes in hepatocellular carcinoma and colorectal liver metastases. Eur J Gastroenterol Hepatol 2008, 20 (5) : 393–398.CrossRefPubMed 27. Tanaka H, Fujita N, Sugimoto R, Urawa N, Horiike S, Kobayashi Y, Iwasa M, Ma N, Kawanishi S, Watanabe S, Kaito M, Takei Y: Hepatic oxidative DNA damage is associated check details with increased risk for hepatocellular carcinoma in chronic hepatitis C. Br J Cancer 2008, 98 (3) : 580–586.CrossRefPubMed 28. Kuramitsu Y, Nakamura K: Proteomic analysis of cancer tissues: Shedding light on carcinogenesis and possible biomarkers.

Proteomics 2006, 6 (20) : 5650–5661.CrossRefPubMed 29. Ezzikouri S, El Feydi AE, Chafik A, Afifi R, El Kihal L, Benazzouz M, Hassar M, Pineau P, Benjelloun S: SRT1720 order Genetic polymorphism in the manganese superoxide dismutase gene is associated with an increased risk for hepatocellular carcinoma in HCV-infected Moroccan patients. Mutat Res 2008, 649 (1–2) : 1–6.PubMed 30. Kuruma H, Egawa S, Oh-Ishi M, Kodera Y, Satoh M, Chen W, Okusa H, Matsumoto K, Maeda T, Baba S: High molecular mass proteome of androgen-independent Thalidomide prostate cancer. Proteomics 2005, 5 (4) : 1097–1112.CrossRefPubMed 31. Tan S, Seow TK, Liang RC, Koh S, Lee CP, Chung MC, Hooi SC: Proteome analysis of butyrate-treated human colon cancer cells (HT-29).

Int J Cancer 2002, 98 (4) : 523–531.CrossRefPubMed 32. Prasannan P, Pike S, Peng K, Shane B, Appling DR: Human mitochondrial C1-tetrahydrofolate synthase: gene structure, tissue distribution of the mRNA, and immunolocalization in Chinese hamster ovary calls. J Biol Chem 2003, 278 (44) : 43178–43187.CrossRefPubMed 33. Howard KM, Muga SJ, Zhang L, Thigpen AE, Appling DR: Characterization of the rat cytoplasmic C1-tetrahydrofolate synthase gene and analysis of its expression in liver regeneration and fetal development. Gene 2003, 319: 85–97.CrossRefPubMed Authors’ contributions NL carried out the 2-DE, participated in MALDI-TOF-MS and drafted the manuscript. YL participated in MALDI-TOF-MS and performed the database analysis. XF is the corresponding author, conceived of the study and designed the study. HL participated in the preparation of tissue protein. CL mainly participated in the database analysis. LC participated in the design of the study and coordination. ZW participated in the collection of liver tissue samples.

NN and MA were supported by the Swiss National Science Foundation grant 31003A_130735. Electronic supplementary material Additional file 1: File S1: Flow cytometry data. (XLS 137 KB) Additional Everolimus file 2: Figure S1: Variation in the expression of ptsG, mglB and rpsM reporters across different environments. The CV of log expression of PptsG-gfp (green), PmglB-gfp (blue) and PrpsM-gfp (red) was plotted against the mean log expression. Power regression was fitted to each dataset corresponding to the expression

of the same reporter across different environments. The individual curves of variation in the expression of ptsG and rpsM reporters showed negative associations between the mean expression and the

variation of expression across environments, whereas the mglB reporter showed a positive association. (TIFF 145 KB) Additional file 3: Text S1: Analysis of expression of fluorescent reporters in glucose-acetate mixtures. (PDF 57 KB) Additional file 4: Figure S2: Reporter expression in mixed-substrate environments. Expression of ptsG, mglB and acs reporters was measured in chemostats (D = 0.15 h-1) in mixed-substrate environments supplemented with 0.28 mM Glc and 0.28 mM Ac (green), or 2.8 mM Glc and 2.8 mM Ac (blue). The distributions were plotted together with the measurements of the reporter expression in the environments see more with only glucose in the feed (0.56 mM Glc – orange, and 5.6 mM

Glc – red). The fluorescence of the promoterless strain is presented in black. (TIFF 544 KB) Additional file 5: Figure S3: Expression of the pck reporter in different chemostat and batch conditions. Ppck-gfp fluorescence (indication of flux to gluconeogenesis) was measured in bacterial populations grown in chemostats (D = 0.15 h-1) and batch environments supplied with minimal media supplemented with only D-glucose, only sodium acetate or D-glucose plus sodium acetate. Again, background fluorescence is the fluorescence of the promoterless strain, depicted in black. The expression of the pck reporter was decreased in the find more exponential phase in glucose batch cultures in comparison Phosphoglycerate kinase to carbon-limited chemostats. (TIFF 757 KB) Additional file 6: Figure S4: Changes in gfp expression prior of reaching theoretical steady-state. Pacs-gfp fluorescence was measured for five independent replicates growing on different concentration of glucose in the feed. At time point of 0 hours, chemostat experiments were started at a minimal dilution rate of D = 0.14 h-1. After 24 hours, dilution rates were increased to D = 0.15 h-1. The fluorescence plots show gfp distribution in bacterial populations without gating, together with fluorescence of the promoterless strain depicted in black. All independent replicates showed reproducible measurements of GFP fluorescence after 3.6 volume turnovers at D = 0.15 h-1.

6 0.14 21.6 1.41 32.48 8.05 40 16.58 3.12 8.78 0.79 81.23 13.55 155 6.36 8.15 0.97 91 5.89 60 34.13 0.58 4.2 0.34 114.39 10.92 264.33 8.14 0 0 45.45 3.67

80 30 1.56 2.78 0.56 236.97 4.73 425.33 8.49 0 0 59.45 6.92 100 50.87 7.17 1.23 0.05 check details 284.6 7.31 590.67 15.56 0 0 37.03 4.78 Conclusions In light of the results reported, both the polymeric concentrations and the deposition method (dipping or spraying) affect the growth of the nanofilms. The roughness obtained with the dipped slides is higher than the registered one with the sprayed substrates; on the other hand, the optical transmittance is lower as a consequence of the greater thickness obtained with the dipped slides. Moreover, in all cases but in the one with 10-3 M of sprayed solutions, the roughness is increased as the number of bilayers grows, which is an unexpected behavior in LbL films. It is also remarkable that the concentrations used here are lower than the ones typically studied in the literature, around 10-2 M [27]. The

thickness and roughness observed using the dipping approach are higher than the ones registered with the sprayed slides: these differences have been observed in previous works [22]. The best results in terms of a superhydrophilic behavior are obtained with 10-3 M dipping solutions and with 10-4 M spraying mixtures. On the other hand, the high optical find more transmittance registered with the 10-4 M of sprayed solutions, even when 100 bilayers are deposited, points to its potential use in applications where superhydrophilic

and transparent surface are required. The use of inorganic short-chain polymers in LbL method shows that some assumed rules need to be redefined. In this work, it has been demonstrated that the roughness of nanofilms can increase as the growing process goes on, depending on the concentration of the polymers used and also on the way during the slides are exposed to the solutions (dipped or sprayed). The highest roughness is obtained when the slides are dipped into the highest concentration solutions, which was supposed to produce the lowest roughness. The thickness of the resulting films falls in the nanometric range so they could be used in applications where surfaces have to be functionalized. Optical transmittance is above 90% for the films prepared with the 10-4 M of sprayed solutions, which highlights its potential used for preparing superhydrophilic transparent films. The use of PSP offers other important advantages: as it is an inorganic polymer, it can yield to surfaces whose degradation is lower than the ones prepared with organic polymers. Therefore, this work enforces to keep on studying the effect of this kind of polymers in LbL nanostructures. Acknowledgements This work was supported by the Spanish Economy and Competitiveness Ministry-FEDER TEC2010-17805. The authors would like to SN-38 concentration express their gratitude to Nadetech Inc. for the design, fabrication, and tune-up of the robot used for the deposition of the nanocoatings.