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Extracellular electron transfer (EET) by the cyanobacterium Microcystis aeruginosa was investigated. Observations indicate that EET onto an electrode poised at + 0.6 vs. standard hydrogen electrode (SHE) is triggered by high pH, more evidently at pH levels above 9. Light intensity does not appear to affect electricity generation, indicating that this may not be a "biophotovoltaic" process. The generated current density was amplified with stepwise pH increases from approximately 5 mA m -2 at pH 7.8 to 30 mA m -2 at pH 10.5, for dense (0.4 mg mL -1 dry weight) Microcystis aeruginosa suspensions with dissolved CO 2 and O 2 approaching equilibrium with atmospheric concentrations. The upsurge in current density was more pronounced (from 5 mA m -2 at pH 7.8 to 40 mA m -2 at pH 10.2) in the absence of the cells' natural electron acceptors, dissolved CO 2 and O 2 . However, the latter effect is more likely due to competition for electrons by oxygen than to reductive stress. EET in this species is therefore a light-independent process that is enhanced by increasing pH, with reasons that are still unknown, but either related to the involvement of protons in the last step of electron transfer, or to intracellular pH control., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Objectives: Family caregiver burden is associated with higher psychological distress. However, little is known about the impact of neighbourhood relationships on caregivers' psychological distress. We examined whether neighbourhood relationships of caregivers moderate the association between family caregiver burden and psychological distress., Study Design: This was a cross-sectional study., Methods: We recruited 5321 Japanese adults who participated in the Japan Multi-Institutional Collaborative Cohort Study in the Okazaki area between 2013 and 2017. Participants completed self-reported questionnaires to measure psychological distress (Kessler 6: K6), subjective caregiver burden, and neighbourhood relationships. We performed a multivariable linear regression analysis in which caregiver burden was designated as an independent variable and the K6 score as a dependent variable, adjusting for demographics. The interaction term between caregiver burden and neighbourhood relationships was also included in the analysis., Results: Data from a total of 5069 participants were included (mean age [standard deviation]: 63.1 years [10.3 years]; 2226 [43.9%] female). Caregiver burden was significantly and positively associated with psychological distress (compared with no burden, mild burden: β = 0.24, P = 0.197; severe burden: β = 0.60, P < 0.01; P for trend < 0.01). There was a significant negative interaction effect of caregiver burden × neighbourhood relationship on psychological distress (severe burden × good neighbourhood relationship: β = -3.29, P < 0.01)., Conclusions: A higher caregiver burden was associated with higher psychological distress, and neighbourhood relationships moderated this association. Our findings suggest that good neighbourhood relationships can buffer caregiving-associated psychological distress., (Copyright © 2020 The Royal Society for Public Health. Published by Elsevier Ltd. All rights reserved.)

Electrochemically polymerized phenothiazines (thionine, methylene green, methylene blue, and toluidine blue) on carbon electrodes were investigated as electron transfer mediators of glucose oxidation by flavin adenine dinucleotide-dependent glucose dehydrogenase (FAD-GDH) for biosensor and biofuel cell applications. Among the tested polyphenothiazines grafted on a glassy carbon electrode, clear redox-mediating activity was observed for poly(methylene green), and the catalytic oxidation current depended on the concentrations of glucose and enzymes and the amount of polymer deposited on the electrode surface. The poly(methylene green)-grafted porous carbon electrodes showed 3 mA cm -2 of glucose oxidation current catalyzed by FAD-GDH., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Background: The objectives of the present study were to assess the mid-term results of cementless total knee arthroplasty (TKA) with the porous tantalum monoblock tibial component and to examine the time course of bone changes on plain radiographs., Methods: The subjects were 32 patients, 29 patients were available for follow-up. We investigated the mid-term results of TKA after a mean follow-up period of 7 years and 8 months. We also examined changes of the bone over time on plain radiographs., Results: The Knee Society Clinical Rating scores showed significant improvement. Bone changes around the tibial component were as follows: new bone formation and longitudinal trabecular thickening in 41.4% (Type A), only longitudinal trabecular thickening in 41.4% (Type B), and no changes in 17.2% (Type C). Type A and B changes were more frequent in patients with osteoarthritis, whereas Type C was only seen in patients with rheumatoid arthritis. Three knees had an initial gap, but this disappeared in all cases, and no new radiolucent lines were detected. Stress shielding was observed in seven knees (21.9%), but there was no implant loosening related to it. When we examined the relationship between the mechanical axis and the locations of the tips of the tibial pegs in patients with or without stress shielding, no significant differences were found., Conclusions: The results of mid-term follow-up have demonstrated favorable bone ingrowth, suggesting that porous tantalum is a promising material for cementless TKA., (© 2013.)

Pentacyanoferrate-bound poly(1-vinylimidazole) (PVI[Fe(CN)5]) was selected as a mediator for amperometric creatinine determination based on the reductive H2O2 detection. Creatinine amidohydrolase (CNH), creatine amidohydrolase (CRH), sarcosine oxidase (SOD), peroxidase (POD), and PVI[Fe(CN)5] were crosslinked with poly(ethylene glycol) diglycidyl ether (PEGDGE) on a glassy carbon (GC) electrode for a creatinine biosensor fabrication. Reduction current was monitored at -0.1V in the presence of creatinine and O2. It is revealed that PVI[Fe(CN)5] is suitable as a mediator for a bioelectrocatalytic reaction of POD, since PVI[Fe(CN)5] neither reacts with reactants nor works as an electron acceptor of SOD. The amounts of PVI[Fe(CN)5], PEGDGE, and enzymes were optimized toward creatinine detection. Nafion as a protecting film successfully prevented the enzyme layer from interferences. The detection limit and linear range in creatinine determination were 12μM and 12-500μM (R(2)=0.993), respectively, and the sensitivity was 11mAcm(-2)M(-1), which is applicable for urine creatinine tests. The results of the creatinine determination for four urine samples measured with this proposed method were compared with Jaffe method, and a good correlation was obtained between the results., (Copyright © 2013 Elsevier B.V. All rights reserved.)

The redox potential of type I copper in the Escherichia coli multicopper oxidase CueO was shifted in the positive or negative direction as a result of the single, double, and triple mutations in the first and second coordination spheres: the formation of the NH···S(-)(Cys500 ligand) hydrogen bond, the breakdown of the NH(His443 ligand)···O(-)(Asp439) hydrogen bond, and the substitution of the Met510 ligand for the non-coordinating Leu or coordinating Gln. Laccase activities of CueO were maximally enhanced 140-fold by virtue of the synergistic effect of mild mutations at and at around the ligand groups to type I copper., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Micro-cubic monolithic carbon cryogel (CCG) electrode was fabricated for the electrochemical fructose dehydrogenase (FDH) reaction. The CCG can adsorb a sufficient amount of FDH even inside the CCG structure. FDH inside the CCG worked well as an electrocatalyst for d-fructose oxidation. An extremely high catalytic current density as large as 0.6A/cm(3) was achieved under stirring condition., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Cyclic voltammograms of yeast extract-containing medium exhibit a clear redox peak around -0.4V vs. Ag|AgCl. Fermentative bacterium Lactococcus lactis was hereby shown to exploit this redox compound for extracellular electron transfer towards a graphite anode using glucose as an electron donor. High performance liquid chromatography revealed that this may be a flavin-type compound. The ability of L. lactis to exploit exogenous flavins for anodic glucose oxidation was confirmed by tests where flavin-type compounds were supplied to the bacterium in well defined media. Based on its mid-point potential, riboflavin can be regarded as a near-optimal mediator for microbially catalyzed anodic electron transfer. Riboflavin derivative flavin mononucleotide (FMN) was also exploited by L. lactis as a redox shuttle, unlike flavin adenine dinucleotide (FAD), possibly due to the absence of a specific transporter for the latter. The use of yeast extract in microbial fuel cell media is herein discouraged based on the related unwanted artificial addition of redox mediators which may distort experimental results., (Copyright 2009 Elsevier B.V. All rights reserved.)

Lactococcus lactis is a gram-positive, normally homolactic fermenter that is known to produce several kinds of membrane associated quinones, which are able to mediate electron transfer to extracellular electron acceptors such as Fe(3+), Cu(2+) and hexacyanoferrate. Here we show that this bacterium is also capable of performing extracellular electron transfer to anodes by utilizing at least two soluble redox mediators, as suggested by the two-step catalytic current developed. One of these two mediators was herein suggested to be 2-amino-3-dicarboxy-1,4-naphthoquinone (ACNQ), via evaluation of standard redox potential, ability of the bacterium to exploit the quinone when exogenously provided, as well as by high performance liquid chromatography coupled with UV spectrum analysis. During electricity generation, L. lactis slightly deviated from its normal homolactic metabolism by excreting acetate and pyruvate in stoichiometric amounts with respect to the electrical current. In this metabolism, the anode takes on the role of electron sink for acetogenic fermentation. The finding that L. lactis self-catalyses anodic electron transfer by excretion of redox mediators is remarkable as the mechanisms of extracellular electron transfer by pure cultures of gram-positive bacteria had previously never been elucidated.

The current density of biofuel cells which use dissolved O(2) as electron acceptor is limited by O(2) supply to the electrode surface due to the low solubility and small diffusion coefficient of O(2) in the electrolyte solution. In order to increase the current density, we constructed an air diffusion biocathode which uses O(2) directly from the air. As cathodic biocatalyst, we utilized CueO from Escherichia coli, which belongs to the family of multi-copper oxidases. O(2) reduction was catalyzed by CueO adsorbed on Ketjen black-modified carbon paper electrodes. The hydrophobic electrode surface was obtained by optimizing the weight ratio of polytetrafluoroethylene binder to Ketjen black. The current density of O(2) reduction reached values as high as 20 mA cm(-2) at 0 V vs. Ag|AgCl, KCl(sat.) in a citrate buffer (1.0 M, pH 5.0, 25 degrees C).

CueO is a multicopper oxidase (MCO) that is involved in the homeostasis of Cu in Escherichia coli and is the sole cuprous oxidase to have ever been found. Differing from other MCOs, the substrate-binding site of CueO is deeply buried under a methionine-rich helical region including alpha-helices 5, 6, and 7 that interfere with the access of organic substrates. We deleted the region Pro357-His406 and replaced it with a Gly-Gly linker. The crystal structures of a truncated mutant in the presence and in the absence of excess Cu(II) indicated that the scaffold of the CueO molecule and metal-binding sites were reserved in comparison with those of CueO. In addition, the high thermostability of the protein molecule and its spectroscopic and magnetic properties due to four Cu centers were also conserved after truncation. As for functions, the cuprous oxidase activity of the mutant was reduced to ca 10% that of recombinant CueO owing to the decrease in the affinity of the labile Cu site for Cu(I) ions, although activities for laccase substrates such as 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), p-phenylenediamine, and 2,6-dimethoxyphenol increased due to changes in the access of these organic substrates to the type I Cu site. The present engineering of CueO indicates that the methionine-rich alpha-helices function as a barrier to the access of bulky organic substrates, which provides CueO with specificity as a cuprous oxidase.

Bioelectrocatalytic oxidation of acetate was investigated under anaerobic conditions by using Escherichia coli K-12 (IFO 3301) cells cultured on aerobic media containing poly-peptone, glucose or acetate as the sole carbon source. It was found that all E. coli cells cultured on the three media work as good catalysts of the electrochemical oxidation of acetate as well as glucose with Fe(CN)6(3-), 2,3-dimethoxy-5-methyl-1,4-benzo-quinone (Q0), 2,6-dichloro-indophenol, or 2-methyl-1,4-naphthoquinone as artificial electron acceptors (mediators). Acetate-grown E. coli cells exhibited the highest relative activity of the acetate oxidation against the glucose oxidation. On the other hand, all the artificial electron acceptors used work as inhibitors for the catalytic oxidation of acetate at increased concentrations. The inhibition phenomenon can be interpreted in terms of competitive substrate inhibition as a whole. Apparent values of Michaelis constant, catalytic constant, and inhibition constant were evaluated by amperometric methods. Q0 is an effective artificial mediator as evidenced by a large reaction rate constant between the cell and Q0 at least at low concentrations (<50 microM). However, Fe(CN)6(3-) is a promising mediator in biosensor applications because the inhibition constant is very large and it works as an electron acceptor even under aerobic conditions.

One-compartment bulk electrolysis and simultaneous spectroscopic measurements are realized in a conventional spectroscopic cuvette without separator by using a mesh-type working electrode with extremely large surface area and a wire-type counter electrode with very small surface area. Spectrophotometric monitoring revealed complete electrolysis in a first-order kinetics. This technique was applied to mediated titration of cytochrome c and bilirubin oxidase for determining their redox potentials. Kinetics for the solution redox reaction between protein and mediator is described. The subtraction of spectral background due to mediator adsorption is very easy because of high reproducibility. The experiments can be done under completely anaerobic conditions. Low-absorbance protein samples (of low concentrations or small absorption coefficients) and hydrophobic proteins (such as membrane-bound proteins) are acceptable for measurements.

We studied vascular endothelial growth factor (VEGF) mRNA synthesis by peripheral blood mononuclear cells (PBMCs) in 30 patients with acute myocardial infarction (AMI) and 20 healthy individuals. PBMCs were isolated from all patients on days 3 and 14 after the onset of aMI, and from all of control individuals. To prepare samples containing identical amounts of GAPDH cDNA, competitive PCR was performed by co-amplifying serial dilutions of GAPDH mutant templates. Next, to measure VEGF cDNA quantitatively in the samples containing identical amounts of GAPDH, we also used competitive PCR by co-amplifying mutant templates of VEGF. The serum VEGF concentrations on day 14 in patients with aMI were measured by an ELISA method. Higher levels of VEGF mRNA in PBMCs were present on day 14 than either on day 3 or in the control group. Serum VEGF concentrations correlated with the VEGF mRNA levels of PBMCs on day 14. Peak serum CK levels correlated well with VEGF mRNA levels of PBMCs on day 14. The present findings suggest that PBMCs may be one of the candidates responsible for elevated circulatory VEGF protein following aMI. In addition, VEGF mRNA may be overexpressed in PBMCs in response to cardiac muscle damage.

We examined serial changes in serum levels of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) measured by ELISAs in 45 patients with acute myocardial infarction (AMI) who received heparin intravenously for 3 to 5 days after the onset and in 30 control subjects with an old myocardial infarction. To evaluate the effect of heparin on these serum levels, heparin was administered intravenously in 10 patients with AMI on day 21. Blood samples were obtained from all AMI patients on days 1, 2, 3, 7, 14, 21, and 28 and from 10 AMI patients before and 1 h after heparin administration. Serum VEGF level was significantly reduced after heparin administration (P<0.001). Serum samples from day 1 to 3 were therefore excluded from the subsequent analysis. Serum VEGF level in AMI patients was significantly higher on day 7 than in the control subjects (P<0.0001), and then decreased over time (P<0.0001). The serum VEGF level on day 7 was independently associated with the peak serum CK level (P<0.05). The serum bFGF level did not differ significantly between the AMI patients and the control subjects. In conclusion, the serum VEGF level may be selectively elevated during the healing process after AMI.

Activity hydrolyzing both N omega-phosphoarginine and glucose-6-phosphate was detected in rat renal microsome but not in hepatic microsome. Renal microsome was solubilized with 1% n-octyl-beta-D-thioglucoside and purified with DEAE-Sepharose column chromatography. Fractions hydrolyzing both N omega-phosphoarginine or glucose-6-phosphate were subjected to 7.5%-polyacrylamide gel electrophoresis in the presence of 0.1% sodium dodecyl sulfate. Phosphatase activity in the gels was detected by a lead nitrate stain using N omega-phosphoarginine or glucose-6-phosphate as substrates. Both substrates produced a stain in the region of the gel corresponding to a protein with a mass of 150 kDa. Extracts of slices from this region of the gel also hydrolyzed phosphocreatine, inorganic pyrophosphate, and O-phosphotyrosine. Moreover, the phosphatase had its optimal pH in the alkaline range and was inhibited completely by 20 microM sodium vanadate, 1 mM cysteine, and 1 mM tetramisole. All these properties indicate that the microsomal phosphoamidase (EC 3.9.1.1) of rat kidney was identical with alkaline phosphatase (EC 3.1.3.1).