Despite the obviously practical value

of peroxidases, at present, their commercial uses are limited, primarily due to its low stability in the presence of hydrogen peroxide, their natural substrate. All heme-proteins, including peroxidases, are inactivated in the presence of some concentrations of hydrogen peroxide. This process, described as a suicide inactivation, is especially important in the absence of reducing substrates, but its mechanism has not been yet fully elucidated [18]. Although the interest to peroxidase started several decades ago, their application as biocatalysts in industrial processes is still negligible due to its inherent instability under operational conditions, mainly caused by the inactivation in the presence HMPL-504 research buy of hydrogen peroxide. The development of techniques for enzyme stabilizing can improve a number of biocatalytic industrial processes. In this work, peroxidase enzyme has been immobilized onto porous silicon (PS) supports for the possible prevention from

its self-inactivation Selleck PLX3397 and its stability under different operational conditions has been analyzed. Methods A commercial peroxidase, Baylase® RP, was kindly donated by Bayer Mexico (Mexico, Federal District, Mexico). Crystalline silicon was a product from Cemat Silicon (Warsaw, Poland). Glutaraldehyde, 3-aminopropyldiethoxysilane, guaiacol, and bovine serum albumin were from Sigma-Aldrich (St. Louis, MO, USA). Bradford reagent was from Bio-Rad (Hercules, CA, USA). All other chemical reagents used in our experiment were of analytical grade

without further purification. Microreactor fabrication Fabrication of porous silicon(PS) <100 > oriented, heavily doped p-type Si wafers with resistivity 0.002 to 0.005 ohm-cm were electrochemically etched with an electrolyte composed of HF/ethanol/glycerol (3:7:1 (v/v)) at a constant current density of 50 mA cm-2 for 170 s to obtain a porous layer of 3,000 ± 60 nm. Functionalization of porous support The porous silicon samples were subjected to thermal oxidation in air at 600°C for 60 min. Silanization process with 3-aminopropyldiethoxysilane (APDES) was performed by immersing the sample in a 5% APDES in toluene for a period of 1 h and annealed at 110°C for 15 min. Glutaraldehyde (GTA, 2.5%) in Molecular motor phosphate buffer pH 6.0 was subsequently coupled to the support for 1 h and finally incubated with peroxidase for 24 h at 4°C. After each step of functionalization, the percent reflectance was measured and the chemical modification of the surface was verified by FTIR. RIFTS, SEM, FTIR, and gravimetric measurement of enzymatic microreactor Reflective interferometric Fourier transform method provides a fast and convenient method of extracting the basic optical parameters modified during the bio-functionalization steps onto of the PS surface.

This later reacts then with substituted hydrazine to give the aminocyanopyrazole 2. Treatment of 2 with orthoester in the presence of catalytic amount of acid furnished the corresponding

cyano-pyrazoloimidates 3 which subsequently were transformed to the corresponding amino pyrazolopyrimidines 4 (Booth et al., 1999; Gupta et al., 2008; Oliveira-Campos et al., 2007; Bakavoli et al., 2010) upon treatment with ammoniac. Reaction of compound 4 with ketene ethoxymethylene compounds 1 in ethanol in presence of catalytic amount of acid furnished the desired 6-cyano-1,7-dihydropyrazolo[3′,4′:4,5]pyrimido[1,6-a]pyrimidine 5a–e in 70 % yield as a yellow solid. The same procedure gave a crystalline ethyl-1,7-dihydro pyrazolo [3′,4′:4,5]pyrimido Selleckchem Entospletinib [1,6-a]pyrimidine-6-carboxylate 5f–i from ethyl-2-cyano-3-ethoxyalkyl-2-enoate in 80 % yield. Scheme 1 shows the synthetic strategy to obtain the target compounds by the four-steps method, yielding the compounds with structure 5a–i listed in Table 1. www.selleckchem.com/products/R406.html Scheme 1

Synthetic procedure of compounds 5a–i. Reagents: i H2N–NHPh, CH3CO2H, CH3CO2H; ii R2C(OEt)3, CH3CO2H; iii NH3; iv Table 1 Synthesis of 7-imino-N 1-phenyl-1,7-dihydro pyrazolo[3′,4′:4,5]pyrimido[1,6-a]pyrimidine 5a–i Compounds R1 R2 R3 Y Yields (%) Reaction time (h) 5a CH3 H H CN 68 24 5b CH3 H CH3 CN 54 71 5c H CH3 H CN 71 24 5d H H H CN 77 5 5e H H C2H5 CN 70 48 5f CH3 H CH3 CO2Et 71 75 5g CH3 H C2H5 CO2Et 69 84 5h H H H CO2Et 89 7 5i H H CH3 CO2Et 78

24 It is interesting Cyclooxygenase (COX) to note that time reaction and yield of products are directly related to the nature of substituent (R3 and Y). The yields of compounds 5h and 5d are 89 and 77 %, respectively. Hydrogen substituent R3 gave superior yields in short time. In all cases, reaction leads to pyrazolo pyrimido pyrimidine only when R1 or R2 is a hydrogen atom. However, steric effect decreased yields of the reaction, as in the case of 5g, and may even prevent the progress of the reaction when R2 and R3 are methyl groups. Analysis of the NMR and IR spectra indicated that compounds 5f–i has ester functional group in their structures so ethoxymethylene cyanoacetate reacts with pyrazolopyrimidine and in both cases Y is CN or CO2Et, nitrogen attacked on the nitrile function as the first attack. Biological activity Anti-inflammatory and gastroprotective activities of compounds 5a, b, f, g The pyrazolopyrimidine derivatives are a well-known class of NSAIDs with several products in market (Russo et al., 1992; El-Kateb et al., 2012) (Figs. 1, 2). Fig. 1 Anti-inflammatory effect of the intraperitoneal administration of 5a, b, f, g and of the reference drug (acetylsalicylic–lysine: ASL) in carrageenan-induced rat paw oedema. The values represent the means difference of volume of paw ± SEM (n = 6). *p < 0.01 and **p < 0.001 significantly different from the control group Fig.

Combining our results with the results from CGM in a previous study, miglitol could reduce glucose fluctuations and hypoglycemic symptoms more effectively than other α-GIs. However, it is still unclear whether glucose fluctuations were lower in type 2 diabetic patients who

were treated longer with miglitol than in those who were treated longer with other α-GIs. Although CGM during the treatment of α-GIs were performed under oral meal loading tests at breakfast, lunch, and dinner in patients hospitalized for 4 days in the previous study [34], the diet during days when SMBG was performed in our selleckchem trials was dependent on each patient. RCT trials, in which dietary habits are well controlled, should examine whether glucose fluctuations by long-term CGM are lower in EPZ-6438 price type 2 diabetic patients treated with miglitol than in those treated with acarbose or voglibose. It should be noted that our trial is a prospective exploratory trial that is not an RCT, which introduces some confounding factors and bias in our trial. It has been reported that blood glucose control is affected by seasonal changes. Indeed, it has been reported that HbA1c has a duration across the year that is highly detected during spring and gradually decreases by autumn in Japan [35]. One of the other possibilities

is that lifestyles such as dietary Selleck Cobimetinib and exercise habits in patients were changed during the trial. In this trial, the doctor assigned caloric intake and the suggestion was not changed during the trial. However,

it is possible that the lifestyles of patients were changed by themselves. In addition, miglitol treatment may reduce a patient’s appetite because the change of α-GI to miglitol treatment inhibits symptoms of hypoglycemia and reduction of blood glucose levels during a meal; however, our results indicate that the change of α-GI to miglitol reduced glucose fluctuation but not HbA1c. Thus, the effect is most likely a result of the effects of miglitol because changes in dietary and exercise habits may alter HbA1c levels. Whether miglitol treatment reduces circulating CVD risk factors including MCP-1 and sE-selectin in type 2 diabetic Japanese patients needs to be examined in an RCT. 5 Conclusion The results of this study indicate that switching from acarbose or voglibose to miglitol for 3 months suppressed glucose fluctuations and serum protein concentrations of MCP-1 and sE-selectin more effectively than the prior α-GI. Acknowledgments This study was sponsored by Sanwa Kagaku Kenkyusho Co., LTD, Nagoya, Japan. Conflict of interest Mr. Fuchigami is an employee of Sanwa Kagaku Kenkyusho Co., LTD, Nagoya, Japan.

Mice were inoculated by intraperitoneal infection with 100 μL of inoculum containing a total of 1 × 105 bacteria (each strain at 5 × 104), consisting of an equal number of wild-type

and mutant strains. At 48 h after infection, the mice were sacrificed by carbon dioxide inhalation. The spleens were homogenized in cold PBS by mechanical disruption. The number of each strain in the spleen was determined by plating a dilution series of the lysate onto LB agar alone and LB agar with appropriate antibiotics. learn more Each competitive index value was calculated as [mutant/wild-type] output/[mutant/wild-type] input and represented as the mean of at least three independent infections. Macrophage survival assay Cells of a mouse macrophage-like line, RAW264.7, PI3K inhibitor were diluted in DMEM containing 10% FBS and seeded in 24-well plates at a density of 5 × 105 cells per well. S. Typhimurium strains were used to infect RAW264.7 cells at a multiplicity of infection of 1. The bacteria were centrifuged onto the cells (500 ×g, 5 min) and incubated for 25 min at 37°C in a 5% CO2 incubator.

Cells were washed three times with PBS, and DMEM containing interferon-γ (IFN-γ) (100 units/well; Peprotech) and gentamicin (100 μg/mL; Sigma) was added. After 95 min of incubation, the medium was replaced with DMEM containing IFN-γ (100 units/well) and gentamicin (10 μg/mL). The number of intracellular bacteria Selleck MG 132 was determined at 2 h and 24 h after infection. For the enumeration of intracellular bacteria, the cells were washed three times with PBS and lysed in 1% Triton X-100, and bacteria were quantified by spreading serial 10-fold dilutions of RAW264.7 cell lysates on LB agar plates to count the colony-forming units (CFU). Each experiment was repeated three times. β-galactosidase assay β-galactosidase activities of reporter gene fusions were determined according to a standard procedure [43]. Statistical analysis The competitive index, mRNA expression,

and bacterial proliferation in macrophage cells were compared using Student’s t-test. For comparative proteomics, the intensity of the spot was compared by one-way ANOVA. Values of P < 0.05 were considered statistically significant. Acknowledgements We thank Toru Hattori (SCRUM inc, Japan) for 2-DE gel image analysis. We thank Kaori Dobashi, Nobue Nameki, Masato Hosono, Kohei Yamashita, and Ayako Mizuta for their technical assistance. This work was supported in part by Grants-in-Aid for Young Scientists (B) (17790291 and 22790415 for TH) and for Scientific Research (C) (17590398 and 21590490 for NO) from the Ministry of Education, Culture, Sports, Science, and Technology of Japan, and by a Kitasato University Research Grant for Young Researchers (2010 for TH). Electronic supplementary material Additional file 1: Table S1. Proteins identified on the reference map. (PDF 101 KB) References 1.

polymyxa M-1 in suppressing E. amylovora and E. carotovora, A-1210477 the causative agents of the important plant diseases fire blight and soft rot, respectively. Since the rare polymyxin P has not been previously used as a clinical agent, in contrast to polymyxin B and colistin [30], this finding provides a potential option to use polymyxin P or its producer strain P. polymyxa M-1 as an alternative of chemical bactericides to control fire blight, soft rot and other plant

diseases caused by gram-negative bacteria. Methods Bacterial strains and growth conditions Strain M-1 isolated from surface sterilized wheat roots in China was kept frozen at −70 C with 15% glycerol as a laboratory stock. This strain was cultured in tryptic soy broth (TSB) liquid medium or on tryptic soy broth

agar (TSBA) plates (TSB supplemented by 1.5% agar) at 30°C for general purposes or in glucose-starch-CaCO3 (GSC) medium [45] at 30°C for antibacterial activity tests MCC950 and chemical analysis of polymyxin. M-1 has been deposited in China General Microbiological Culture Collection Center (CGMCC) as strain CGMCC 7581. Other strains used in this study were laboratory stocks obtained from different sources and kept frozen with 15% (v/v) glycerol at −70°C. They were grown in Luria broth (LB) or on LB agar plates (LB solidified with 1.5% agar) at 30°C (E. amylovora Ea273, E. carotovora and Micrococcus luteus) or 37°C (Pseudomonas aeruginosa, Streptococcus faecalis, Bacillus

megaterium, Bacillus subtilis 168, Bacillus amyloliquefaciens FZB42 and Bacillus cereus ATCC 14579). Bacterial identification Identification of the strain M-1 was carried out by using 16S rDNA sequence analysis as well as by physiological and biochemical characterization. After growing in TSB medium at 30°C overnight, the bacteria cells were collected by centrifuging for chromosomal DNA isolation using the standard phenol:chloroform procedure. Then, the 16S rDNA was amplified by PCR with two pairs of primers 63 F (5’CAG GCC TAA CAC ATG CAA GTC-3’), 1387R (5’GGG CGG TGA TGT ACA AGG C’-3) [46], 530 F (5’GTG CCA GCM GCC GCG G-3’) and 1494R Inositol monophosphatase 1 (5’GGY TAC CTT GTT ACG ACT T-3’) [46, 47]. The reaction mixture included Taq DNA polymerase, 10 × Taq buffer, forward and reverse primers, each deoxynucleoside triphosphate (dATP, dGTP, dCTP and dTTP) (Beijing Youbo Gene Technology Co., Ltd) and template DNA. Amplifications were performed using a Biometra T personal 48 thermocycler (Biometra, Goettingen, Germany) with the following cycle conditions: initial activation at 94°C for 5 min; 35 cycles of 94°C for 1 min, 55°C for 30 sec, and 72°C for 1 min; a final extension at 72°C for 10 min. PCR products (100 μL total volume) were analyzed by electrophoresis using a 0.8% (w/v) Tris-acetate-EDTA (TAE) agarose gel mixed with ethidium bromide and ultraviolet visualization.

Passlick B, Pantel K, Kubuschok B, Angstwurm M, Neher A, Thetter O, Schweiberer L, Izbicki JR: Expression of MHC molecules and ICAM-1 on non-small cell lung carcinomas: association with early lymphatic spread of tumour cells. Eur J Cancer 1996, 32A:141–145.PubMed 26. Vitale M, Rezzani R, Rodella L, Zauli G, Grigolato P, Cadei M, Hicklin DJ, Ferrone S: HLA class I antigen and

transporter associated with antigen processing (TAP1 and TAP2) down-regulation in high-grade primary breast carcinoma lesions. Cancer Res 1998, 58:737–742.PubMed 27. Saio M, Teicher M, Campbell G, Feiner H, Delgado Y, Frey AB: Immunocytochemical demonstration of down regulation of HLA class-I molecule expression in human metastatic breast carcinoma. Clin Exp Metastasis 2004, 21:243–249.PubMed 28. Ryschich E, Notzel T, Hinz U, Autschbach F, Ferguson Alisertib in vivo J, Simon I, Weitz J, Frohlich B, Klar E, Buchler MW, Schmidt J: Control of T-cell-mediated immune response by HLA class I in human pancreatic carcinoma. Clin Cancer Res 2005,11(2 Pt 1):498–504.PubMed 29. Sharpe JC, Abel PD, Gilbertson JA, Brawn P, Foster CS: Modulated expression of human leucocyte antigen class I and class II determinants in hyperplastic and malignant human this website prostatic epithelium. Br J Urol 1994, 74:609–616.PubMed 30. Brasanac D, Markovic-Lipkovski J, Hadzi-Djokic J, Muller GA, Muller CA: Immunohistochemical analysis of HLA class II antigens and tumor infiltrating mononuclear cells in renal cell carcinoma:

correlation with clinical and histopathological data. Neoplasma 1999, 46:173–178.PubMed 31. Hilders CG, Houbiers JG, van Ravenswaay Claasen HH, Veldhuizen RW, Fleuren GJ: Association between HLA-expression and infiltration of immune cells in cervical carcinoma. Lab Invest 1993, 69:651–659.PubMed 32. Hilders CG, Munoz IM, Nooyen Y, Fleuren GJ: Altered HLA expression by metastatic cervical carcinoma cells as a factor in impaired immune surveillance. Gynecol Oncol 1995, 57:366–375.PubMed 33. Cruz I, Meijer CJ, Walboomers JM, Snijders PJ, Van der Waal I: Lack of MHC class I surface expression on neoplastic cells and poor activation of the secretory pathway of cytotoxic cells in oral squamous cell carcinomas.

Br J Cancer 1999, 81:881–889.PubMed 34. Grandis JR, Falkner DM, Melhem MF, Gooding WE, Drenning SD, Morel Urease PA: Human leukocyte antigen class I allelic and haplotype loss in squamous cell carcinoma of the head and neck: clinical and immunogenetic consequences. Clin Cancer Res 2000, 6:2794–2802.PubMed 35. Gati A, Da Rocha S, Guerra N, Escudier B, Moretta A, Chouaib S, Angevin E, Caignard A: Analysis of the natural killer mediated immune response in metastatic renal cell carcinoma patients. Int J Cancer 2004, 109:393–401.PubMed 36. Lanier LL: Natural killer cells: from no receptors to too many. Immunity 1997, 6:371–378.PubMed 37. Doubrovina ES, Doubrovin MM, Vider E, Sisson RB, O’Reilly RJ, Dupont B, Vyas YM: Evasion from NK cell immunity by MHC class I chain-related molecules expressing colon adenocarcinoma.

PubMedCrossRef 34. Chattopadhyay S, Fensterl V, Zhang Y, Veleeparambil M, Yamashita M, Sen GC: Role of interferon regulatory factor 3-mediated apoptosis in the establishment and maintenance of persistent infection by Sendai virus. J Virol 2013, 87:16–24.PubMedCentralPubMedCrossRef

35. Uslu R, Sanli UA, Sezgin C, Karabulut B, Terzioglu E, Omay SB: Arsenic trioxide-mediated cytotoxicity and apoptosis in prostate and ovarian carcinoma cell lines. Clin Cancer Res 2000, 6:4957–4964.PubMed Tanespimycin chemical structure 36. Jang M, Kim Y, Won H, Lim S, KRJ , Dashdorj A, Min YH, Kim SY, Shokat KM, Ha J, Kim SS: Carbonyl reductase 1 offers a novel therapeutic target to enhance leukemia treatment by arsenic trioxide. Cancer Res 2012, 72:4214–4224.PubMedCrossRef 37. Chen GQ, Shi XG, Tang W, Xiong SM, Zhu J, Cai X, Han ZG, Ni JH, Shi GY, Jia PM, Liu MM, He KL, Ma J, Zhang P, Zhang TD, Paul P, Naoe T, Kitamura K, Miller W, Waxman S, Wang ZY, de The H, Chen SJ, Chen Z: Use of arsenic trioxide (As 2 O 3 ) in the treatment of acute promyelocytic STI571 nmr leukemia (APL): I. As 2 O 3 exerts dose-dependent dual effects on APL cells. Blood 1997, 89:3345–3353.PubMed 38. Ma DC, Sun YH, Chang KZ, Ma XF, Huang SL, Bai YH, Kang J, Liu YG, Chu JJ: Selective induction of apoptosis of NB4 cells from G2 + M phase by sodium arsenite at lower doses. Eur J Haematol 1998, 61:27–35.PubMedCrossRef 39. Baysan A, Yel L, Gollapudi S, Su H, Gupta S: Arsenic trioxide induces apoptosis via the mitochondrial pathway

by upregulating the expression of Bax and Bim in human B cells. Int J Oncol 2007, 30:313–318.PubMed 40. Kang YH, Lee SJ: The role of p38 MAPK and JNK in arsenic trioxide-induced mitochondrial cell death in human cervical OSBPL9 cancer cells. J Cell Physiol 2008, 217:23–33.PubMedCrossRef 41. Catalani S, Carbonaro V, Palma F, Arshakyan M, Galati R, Nuvoli B, Battistelli S, Canestrari F, Benedetti S: Metabolism modifications and apoptosis induction after Cellfood™ administration to leukemia cell lines. J Exp Clin Cancer Res 2013, 32:63.PubMedCentralPubMedCrossRef 42. Niero EL, Machado-Santelli GM: Cinnamic acid induces apoptotic cell death and cytoskeleton disruption in human

melanoma cells. J Exp Clin Cancer Res 2013, 32:31.PubMedCentralPubMedCrossRef 43. Huang Y, Hu J, Zheng J, Li J, Wei T, Zheng Z, Chen Y: Down-regulation of the PI3K/Akt signaling pathway and induction of apoptosis in CA46 Burkitt lymphoma cells by baicalin. J Exp Clin Cancer Res 2012, 31:48.PubMedCentralPubMedCrossRef 44. Okui T, Fujiwara Y: Inhibition of human excision DNA repair by inorganic arsenic and the comutageniceffect in V79 Chinese hamster cells. Mutat Res 1986, 172:69–76.PubMedCrossRef 45. Kryeziu K, Jungwirth U, Hoda MA, Ferk F, Knasmüller S, Karnthaler-Benbakka C, Kowol CR, Berger W, Heffeter P: Synergistic anticancer activity of arsenic trioxide with erlotinib is based on inhibition of EGFR-mediated DNA double-strand break repair. Mol Cancer Ther 2013, 12:1073–1084.PubMedCrossRef 46.

Actinic intensity was increased in ten steps from 10 to 1,600 μmol quanta m−2 s−1 of 635 nm light. Leaf pre-illuminated for 1 h at 600 μmol m−2 s−1, with 10 min dark time before start of recording. Screenshot of the original recording (Dual-PAM user software). b Deconvolution of the ΔpH and ΔΨ components of the overall pmf by the DIRK method. Zoomed detail of the data set presented in a, showing dark-interval relaxation kinetics after turning off 200 μmol m−2 s−1 (light step 5 in a). c Partitioning of overall proton motive force (pmf)

into ΔpH and ΔΨ components as a function of light intensity during the course of the experiment depicted in a. ΔpH and ΔΨ were determined as explained in b As has been discussed extensively CAL-101 mouse by Kramer and co-workers (for reviews see Kramer et al. 2004a, b; Cruz et al. 2004; Avenson et al. 2005b), the pmf and its ΔpH and ΔΨ components play a dual role in photosynthesis, namely at the level of energy transduction (synthesis of ATP from ADP click here and Pi at the thylakoid CF0–CF1 ATP synthase) and at the level of regulation. In particular, the ΔpH has been known to regulate the efficiency of light capture in PS II via dissipation of excess energy, which otherwise would lead to photodamage (Demmig-Adams

1992; Niyogi 1999). The observed increase of the ΔpH component above 300 μmol m−2 s−1 on the cost of the ΔΨ component (Fig. 2c) may serve as an example for the adaptive flexibility Niclosamide of the photosynthetic apparatus. While ΔΨ contributes substantially to overall pmf at moderate PAR, where the efficiency of light capture is decisive, maximal ΔpH is approached at high light intensities only, where down-regulation of PS II becomes essential. Very recently Johnson and Ruban (2013) questioned the existence of a substantial ΔΨ components in plant

leaves during steady-state illumination, as suggested by Kramer and co-workers, on the grounds of experiments with nigericin-infiltrated leaves of wild-type Arabidopsis and with leaves of Arabidopsis mutants deficient in energy-dependent fluorescence quenching (qE). These authors argue that the apparent ECS in normal leaves during steady-state illumination is not due to a genuine 515 nm change, i.e., is not caused by ΔΨ, but in fact reflects an overlapping qE-related absorption change, the position of which varies depending on the xanthophyll content of the leaves between 525 and 540 nm (Johnson et al. 2009). It may be pointed out that all measurements of Johnson and Ruban (2013) were carried out using 700 μmol m−2 s−1 red light, i.e., at a high intensity of absorbed light, where also our data show a rather small ΔΨ component (Fig. 2c).

The CPE was introduced instead of a pure capacitor in the simulations to obtain a good agreement between the experimental and simulation data. The CPE impedance can be defined

as Z CPE = Q -1.(jω)-n where “n” is related to the slope of log Z vs. log f in the Bode plot, ω is the angular frequency and Q is the combination of properties related to both the surface and the electro-active species, and is independent of frequency. The CPE depends on both the parameter Q and the exponent “n,” but it should be stressed that Q is often approximated to capacitance. The CPE is in parallel arrangement with R 2-W elements, where R 2 is the charge transfer resistance which is in series with the Warburg element W. The circuit diagram is consistent with earlier results using the [FeII(CN)6]4- Linsitinib purchase / [FeIII(CN)6]3- redox couple in solution [13]. For a simple parallel resistance-capacitance combination, the conversion of the CPE parameter into capacitance can be estimated from the following equation [31]: where C is the capacitance and ω m,I = 2πf and f is the frequency at which the imaginary impedance Z I is maximum, and Q is the CPE parameter. Table 1 shows the results from the simulation experiments for both GO and ERGO. It can be seen that ERGO has lower charge transfer resistance compared to GO, which is consistent with previous works [13], where the

charge transfer resistance of ERGO and GO is 333 and 831 Ω·cm2, respectively. The charge transfer resistance XMU-MP-1 price of ERGO reported by Pumera [13] was deposited from GO at a constant potential of -1.2 V vs. Ag/AgCl in phosphate buffer solution at pH 7.2. The ID/IG peak for ERGO and GO obtained in this work in the “FTIR and Raman spectra” section is lower than previous

report [13], and the FTIR results also shows the presence of the sp2 hybridized C=C at around 1,610 cm-1 which could explain the lower charge transfer resistance in this work. Clearly, the electrolyte medium and the experimental conditions greatly influenced the charge transfer resistance value of ERGO. This higher charge transfer resistance of ERGO is primarily due to its higher electrical conductivity [32]. The chemical reduction of GO using sodium hydrosulfite nearly to produce RGO also gave an electrical conductivity of seven orders of magnitude higher than GO [33]. The higher electrical conductivity of ERGO could facilitate faster electron transfer to the [FeII(CN)6]4-/[FeIII(CN)6]3- redox couple, thus ERGO has lower charge transfer resistance R2 compared to GO. The higher charge transfer resistance of GO compared to ERGO in Table 1 has a good correlation with the higher electrical resistivity of GO compared to RGO obtained by Zhou [33]. It can be seen also that the value of surface capacitance for ERGO is nearly five times higher compared to that for GO.

Quinone species were identified by their spectrum and the equivalent number of isoprene units (Hiraishi et al. 1989). Acid volatile sulfides Sediment samples were collected at up to ~10 cm depth from surface layer of all sites on 20

and 21 January 2011, and the concentration of acid volatile sulfides (AVS) in the sediments was determined in triplicate using an AVS detector tube (210H and 210L, Gastec, Ayase, Japan) following the manufacturer’s instructions. Statistical analyses Microbial dissimilarity To investigate the quantitative differences KU 57788 in the microbial community structure based on respiratory quinone in the sediments, a dissimilarity index value (D-value) was calculated using Eq. (1) (Hiraishi et al. 1991): $$ D\left( i,j \right) =

\frac12\sum\limits_k = 1^n , $$ (1)where n is the number of quinone species and f i,k and f j,k are AZD9291 the molar fractions of quinone species k for any two samples i and j, respectively. The D-value ranged from 0 to 1. The values greater than 0.2 were interpreted as having a significant difference in the microbial community (Hiraishi et al. 1991). To visually understand microbial dissimilarity among all the sediment samples, multidimensional scaling (MDS) and cluster analysis with an unweighted pair group method using arithmetic averages were carried out on the basis of the quinone fraction using a statistical package (PASW® Statistics CYTH4 18, SPSS Japan, Tokyo, Japan). The Kruskal’s

stress and R 2 measures are used to test the reliability and validity of the MDS results; Kruskal’s stress is the measure most commonly used for determining the MDS model’s badness of fit. Kruskal and Wish (1978) give the following numbers as guideline: 0.00 a perfect fit, 0.025 an excellent fit, 0.05 a good fit, 0.10 a fair fit and 0.20 a poor fit. An R 2 of 0.6 is considered the minimum acceptable level for the validity of the MDS analysis. Microbial diversity To evaluate microbial diversity in terms of the richness and evenness of the quinone species, Shannon–Wiener diversity H′ was estimated according to Eq. (2) (Shannon and Weaver 1963): $$ H^\prime = – \sum\limits_k = 1^n \left( f_k \ln f_k \right) , $$ (2)where n is the number of quinone species and f k is the molar fraction of quinone species k for a sample. Typically, the value ranged from 1.5 to 3.5, indicating a low to high richness and evenness of species. Results and discussion Water pollution status Water quality Average EC and salinity at site 1 were 52.8 mS/cm and 34.7 ‰, respectively, which are comparable to values of natural seawater (Fig. 3). A temporary drop in EC and salinity was found at about 0800 hours on 6 April because of rainfall. The values at sites 2-2 and 3 were slightly lower than those values at site 1 and then lower than those of natural seawater.