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Aims: To study the effect of pH, temperature and substrate on the magnitude of N2O and NO production by heterotrophic nitrifiers. Methods and Results: The change in N2O and NO production by the heterotrophic nitrifiers Alcaligenes faecalis subsp. parafaecalis and Paracoccus pantotrophus because of variations in pH, temperature and substrate was studied in chemostat cultures under steady-state conditions. N2O, NO and CO2 production increased with temperature between 4 and 32°C. For N2O an optimum temperature of 28°C was observed. No optimum temperature was found for NO. Highest N2O and CO2 productions were observed at a pH of 7·0. However, besides having an optimum at pH 7·0, especially NO production but also N2O production increased significantly at pH ≤ 4·0. This increase in NO production under acidic conditions was partly because of chemo-denitrification, which contributed up to 62% of total NO production at pH 3·0 (0·8% for N2O). Furthermore, we could demonstrate that substrate quality significantly affects N2O, NO and CO2 production. N2O and especially NO production by A. faecalis p. was significantly lower on an ammonium citrate medium when compared with rates obtained for a peptone-meat extract medium. Conclusions: The results indicate that heterotrophic nitrifiers are suitable model organisms to study the influence of environmental factors on microbial N trace gas production. Significance and Impact of the Study: The results allow an improved description, e.g. of the pH dependency of N trace gas production by microbes and/or chemo-denitrification in process-oriented models describing the exchange of N trace gases between soils and the atmosphere. [ABSTRACT FROM AUTHOR]

Production of NO and N2O by the heterotrophic nitrifier Alcaligenes faecalis subsp. parafaecalis was studied during growth in batch and continuous culture on peptone-meat extract medium. Depending on oxygen saturation level, medium redox status and amount of substrate supplied, the microorganisms produced 0.002–0.25 mg NO-N h- 1 (g protein)- 1 and 0.16–2.4 mg N2O-N h- 1 (g protein)- 1. Maximum rates of nitrogen oxides production were observed during peak events initiated by sudden changes of oxygen supply in the medium and were due to combined nitrification/denitrification taking place simultaneously within the cells. Based on model simulations of enzymatic kinetics of denitrification, possible mechanisms of increased nitrogen oxides production during periods of changes in oxygen supply are suggested. [ABSTRACT FROM AUTHOR]

Presents a response by M. Kesik et al to the comments on their study "Inventories of N2 and NO Emissions From European Forest Soils," which appeared in the 2005 issue of "Biogeosciences Discussions." Variability of soil pH on microsite scales; Site differences caused by differences in the forest floor humus type.

A novel flexible glucose biosensor using vertically aligned carbon nanotubes (VACNT) and a conjugated polymer (CP) was fabricated. A scaffold based on VACNT grown on aluminum foil (VACNT-Al foil) with poly (9,9-di-(2-ethylhexyl)-fluorenyl-2,7-diyl)-end capped with 2,5-diphenyl-1,2,4-oxadiazole (PFLO) was used as the immobilization matrix for the glucose biosensor. Glucose oxidase (GOx) was immobilized on a modified indium tin oxide (ITO) coated polyethylene terephthalate (PET) electrode surface. The biosensor response at a potential of -0.7V versus Ag wire was followed by the decrease in oxygen level as a result of enzymatic reaction. The biosensor exhibited a linear range between 0.02mM and 0.5mM glucose and kinetic parameters (K M app , I max , limit of detection (LOD) and sensitivity) were estimated as 0.193mM, 8.170μA, 7.035×10 -3 mM and 65.816μA/mMcm 2 , respectively. Scanning electron microscopy (SEM) was used for surface characterization. The constructed biosensor was applied to determine the glucose content in several beverages., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

A novel glucose oxidase (GOx) based amperometric biosensor utilizing a conducting polymer (CP), multi walled carbon nanotubes (MWCNTs) and a novel water soluble zinc phthalocyanine (ZnPc) was constructed. For this purpose, a novel ZnPc was synthesized to examine the role of being a part of support material for enzyme deposition. High water solubility was achieved with the introduction of tetra quaternized imidazolyl moieties at the peripheral positions of phthalocyanine. In order to fabricate the proposed biosensor, a graphite electrode was firstly modified with poly[9,9-di-(2-ethylhexyl)- fluorenyl-2,7-diyl] end capped with N,N-Bis(4- methylphenyl)-4-aniline (PFLA) and MWCNTs. Then, GOx was co-immobilized with ZnPc onto the modified surface. To the best our knowledge, a sensor design which combines conjugated polymer/MWCNTs/ZnPc was attempted for the first time and this approach resulted in improved biosensor characteristics. The constructed biosensor showed a linear response for glucose between 0.025-1.0mM with a detection limit of 0.018mM. K M app and sensitivity values were calculated as 0.53mM and 82.18μAmm -1 cm -2 , respectively. Moreover, scanning electron microscopy (SEM) and cyclic voltammetry (CV) techniques were used to investigate the surface modifications. Finally, fabricated biosensor was tested on beverages for glucose detection successfully., (Copyright © 2016 Elsevier B.V. All rights reserved.)

To construct a sensing interface, in the present work, a conjugated polymer and core-shell magnetic nanoparticle containing biosensor was constructed for the pesticide analysis. The monomer 4,7-di(furan-2-yl)benzo[c][1,2,5]thiadiazole (FBThF) and core-shell magnetic nanoparticles were designed and synthesized for fabrication of the biosensing device. The magnetic nanoparticles were first treated with silica and then modified using carboxyl groups, which enabled binding of the biomolecules covalently. For the construction of the proposed sensor a two-step procedure was performed. First, the poly(FBThF) was electrochemically generated on the electrode surface. Then, carboxyl group modified magnetic nanoparticles (f-MNPs) and acetylcholinesterase (AChE), the model enzyme, were co-immobilized on the polymer-coated surface. Thereby, a robust and novel surface, conjugated polymer bearing magnetic nanoparticles with pendant carboxyl groups, was constructed, which was characterized using Fourier transform infrared spectrometer, cyclic voltammetry, scanning electron microscopy, and contact angle measurements. This novel architecture was then applied as an immobilization platform to detect pesticides. To the best of our knowledge, a sensor design that combines both conjugated polymer and magnetic nanoparticles was attempted for the first time, and this approach resulted in improved biosensor characteristics. Hence, this approach opens a new perspective in the field of enzyme immobilization and sensing applications. Paraoxon and trichlorfon were selected as the model toxicants. To obtain best biosensor performance, optimization studies were performed. Under optimized conditions, the biosensor in concern revealed a rapid response (5 s), a low detection limit (6.66 × 10(-3) mM), and high sensitivity (45.01 μA mM(-1) cm(-2)). The KM(app) value of poly(FBThF)/f-MNPs/AChE were determined as 0.73 mM. Furthermore, there was no considerable activity loss for 10 d for poly(FBThF)/f-MNPs/AChE biofilm.

A novel amperometric pyranose oxidase (PyOx) biosensor based on a selenium containing conducting polymer has been developed for the glucose detection. For this purpose, a conducting polymer; poly(4,7-bis(thieno[3,2-b]thiophen-2-yl)benzo[c][1,2,5] selenadiazole) (poly(BSeTT)) was synthesized via electropolymerisation on gold electrode to examine its matrix property for glucose detection. For this purpose, PyOx was used as the model enzyme and immobilised via physical adsorption technique. Amperometric detection of consumed oxygen was monitored at -0.7 V vs Ag reference electrode in a phosphate buffer (50 mM, pH 7.0). K(M)(app), Imax, LOD and sensitivity were calculated as 0.229 mM, 42.37 nA, 3.3 × 10(-4)nM and 6.4 nA/mM cm(2), respectively. Scanning electron microscopy (SEM), Electrochemical Impedance Spectroscopy (EIS) and cyclic voltammetry (CV) techniques were used to monitor changes in surface morphologies and to run electrochemical characterisations. Finally, the constructed biosensor was applied for the determination of glucose in beverages successfully., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

The combination of supramolecules and conducting polymers (CPs) has gained much attention for the development of new immobilization matrices for biomolecules. Herein, an amperometric biosensor based on a novel conducting polymer, poly(2-(2-octyldodecyl)-4,7-di(selenoph-2-yl)-2H-benzo[d][1,2,3]triazole)) (PSBTz) and β-cyclodextrin (β-CD) for the detection of cholesterol, was constructed. The PSBTz film with β-CD was deposited on a graphite electrode by electropolymerization technique to achieve a suitable matrix for enzyme immobilization. Moreover, to justify the immobilization, alkyl chain containing conducting polymer (PSBTz) was designed, synthesized and electrochemically polymerized on the transducer surface. Alkyl chains in the structure of SBTz and hydroxyl groups of β-CD contributed to effective immobilization while protecting the suitable orientation of the biomolecule. Cholesterol oxidase (ChOx) was covalently immobilized onto the modified surface using N,N′-carbonyldiimidazole (CDI) as the cross-linking agent. After successful immobilization, amperometric biosensor responses were recorded at −0.7 V vs Ag/AgCl in phosphate buffer (pH 7.0). The apparent Michaelis-Menten constant (KM(app)), maximum current (Imax), limit of detection (LOD), and sensitivity values were determined: 28.9 μM, 12.1 μA, 0.005 μM, and 5.77 μA/μM cm(2), respectively. The fabricated biosensor was characterized using scanning electron microscopy (SEM) and cyclic voltammetry (CV) techniques. Finally, the prepared biosensor was successfully applied for the determination of cholesterol in blood samples.

We describe a modification and post-functionalization technique for a donor-acceptor-donor type monomer; 6-(4,7-bis(2,3-dihydrothieno[3,4-b][1,4]dioxin-5-yl)-2H-benzo[d][1,2, 3]triazol-2-yl)hexan-1-amine. Folic acid was attached to the fluorescent structure. The conjugation was confirmed via NMR and Fourier transform infrared analyses. Cytotoxicity was investigated and the comparison of association of targeted monomeric structures in tumor cells was monitored via fluorescence microscopy., (© 2014 American Institute of Chemical Engineers.)

Molecular dynamics simulations are performed to determine the solubility and diffusion coefficient of carbon dioxide and nitrogen in poly(methyl methacrylate) (PMMA). The solubilities of CO2 in the polymer are calculated employing our grand canonical ensemble simulation method, fixing the target excess chemical potential of CO2 in the polymer and varying the number of CO2 molecules in the polymer matrix till establishing equilibrium. It is shown that the calculated sorption isotherms of CO2 in PMMA, employing this method well agrees with experiment. Our results on the diffusion coefficients of CO2 and N2 in PMMA are shown to obey a common hopping mechanism. It is shown that the higher solubility of CO2 than that of N2 is a consequence of more attractive interactions between the carbonyl group of polymer and the sorbent. While the residence time of CO2 beside the carbonyl group of polymer is about three times higher than that of N2, the diffusion coefficient of CO2 in PMMA is higher than that of N2. The higher diffusion coefficient of CO2, compared to N2, in PMMA is shown to be due to the higher (≈3 times) swelling of polymer upon CO2 uptake.

Cysteine proteinases released by Fasciola hepatica play a key role in parasite feeding, migration through host tissues and in immune evasion. Hence, a recombinant cysteine proteinase (CPFhW) expressed as inclusion bodies in Escherichia coli was used for enteral vaccination of rats against fasciolosis. We managed to activate this proteinase and found it to have cathepsin L1-like substrate preference. Enteral vaccination of rats induced a 78-80% protection against challenge with fluke metacercariae (mc). The immunised rats showed clear immunological response. The challenge with mc caused a remarkable infiltration of eosinophils into the peritoneal cavity of both the vaccinated rats and challenge control rats. However, CD8+ and CD4+ lymphocytes appeared in significantly higher numbers in the peritoneal fluid of vaccinated rats than in controls.

A fragment of non-glycosylated E2 antigen of classical swine fever virus (CSFV), lacking the trans-membrane anchor (TM-) of the native glycoprotein, was produced in recombinant Escherichia coli strain BL21(DE3) in the form of inclusion bodies. These inclusion bodies isolated from the bacteria cells were administrated orally to mice twice at either 10 or 50 microg per dose. Each mouse fed with inclusion bodies carrying the E2 antigen responded with plasma antibodies and/or fecal IgA at least once during the entire investigation. Our study showed the capacity of inclusion bodies to induce both systemic and mucosal responses as well as to evoke relatively-long mucosal memory when fed to mice at low-number vaccination schedule and without any adjuvant. We propose the use of inclusion bodies for oral vaccination as an alternative to artificial systems for delivery of recombinant antigens by the oral route. Very few steps are needed to obtain an antigen ready for use as a vaccine. The procedure is easy and inexpensive and can be used for development of vaccine against classical swine fever.