Synthesis, Characterization, and Cytotoxicity Analysis of a Biodegradable Polyurethane

A polyester urethane was synthesized for use in a biodegradable scaffold. The polyurethane was synthesized in a two-step process: first, ester diol was synthesized from lactic acid and polyethylene glycol 400 (PEG 400), then it was polymerized with toluene diisocyanate using dibutyl tin dilaurate (DBTDL) as a catalyst to form a polyester urethane. Polyester urethane has tensile strength of 51-59 MPa and elongation at fracture of 369-439%. FTIR and XRD were used to confirm the formation and structure of the polymer. Hydrolytic degradation was studied in different alkali solutions and in saline water. In order to assess the cellular response of this material, cytotoxicity analysis was carried out against the cell line.     

Crystallization and dissolution behavior of l(+) calcium and zinc lactate in ethanol-water solvent

Crystallization and dissolution behavior of L(+) calcium and zinc lactate in 50% ethanol-water solution was studied. The effects of stirring, standing, ultrasonic wave, and temperature on the crystallization of L(+) calcium and zinc lactate were evaluated. It was found that standing had a positive effect on crystallization of L(+) calcium and zinc lactate, while stirring promoted dissolution of crystallized particles and resulted in high residual concentration in mother liquor. The application of ultrasonic wave did not influence much on crystallization process. L(+) calcium and zinc lactate crystallized easily at 5 ‡C; however, complete crystallization took more than 72 hours. These two salts dissolved rapidly and reached equilibrium within 1 hour.     

The effect of hyperoxia on submaximal exercise with the self-contained breathing apparatus

The e^ffects of hyperoxia on submaximal exercise with the self-contained breathing apparatus (SCBA) were studied in 25 males. Each participant completed a graded exercise test for the determination of ventilatory threshold (VT) and then a submaximal practice trial with a normoxic gas mixture. The normoxic (20.93 ± 0.22% O₂ ; SUB₂₁) and hyperoxic (40.18 ± 0.73% O₂; SUB₄₀) submaximal trials were then administered in a random order. All exercise tests were completed on separate days while wearing firefighting gear and the SCBA. Compared with SUB₂₁, hyperoxia significantly reduced minute ventilation ([Vdot]_E ), mask pressure (P_mask), heart rate, blood lactate concentration, perceived exertion, and perceived breathing distress. As expected, hemoglobin saturation remained higher (p<0.05) during SUB₄₀. The reductions in both [Vdot]_E and P_mask with hyperoxia imply a reduction in the work of breathing during exercise. Total gas consumption was 10.3 ± 8.1% lower during SUB₄₀ when compared to SUB₂₁, another finding that has significant practical implications for occupational safety.     

Redox-flexible NADH oxidase biosensor: A platform for various dehydrogenase bioassays and biosensors

A generic amperometric bioassay based on the enzymatic oxidation catalysed by the stable NADH oxidase (NAox) from Thermus thermophilus has been developed for NADH measurements. The NAox uses O₂ as its natural electron acceptor and produces H₂O₂ in a two-electron process. Electrochemical and spectrophotometric experiments showed that the NAox used in this work, presents a very good activity towards its substrate and, in contrary to previously mentioned NADH oxidases, does not require the addition of any exogenous flavin cofactor neither to promote nor to maintain its activity. In addition, the NAox used also works with artificial electron acceptors like ferrocene derivatives. O₂ was successfully replaced by redox mediators such as hydroxymethyl ferrocene (FcCH₂OH) for the regeneration of the active enzyme. Combining the NAox with the mediator and the horseradish peroxidase we developed an original, high sensitive “redox-flexible” NADH amperometric bioassay working in a large window of applied potentials in both oxidation and reduction modes. The biosensor has a continuous and complementary linearity range permitting to measure NADH concentrations starting from 5 × 10⁻⁶ M in reduction until 2 × 10³ M in oxidation. This redox-flexibility allows choosing the applied potential in order to avoid electrochemical interferences. The association of the “redox-flexible” concept with NADH dependent enzymes opens a novel strategy for dehydrogenases based bioassays and biosensors. The great number of dehydrogenases available makes the concept applicable for numerous substrates to analyse. Moreover it allows the development of a wide range of biosensors on the basis of a generic platform. This gives several advantages over the previous manufacturing techniques and offers a general and flexible scheme for the fabrication of biosensors presenting high sensitivities, wide calibration ranges and less affected by electrochemical interferences.     

Self-containing reactant biochips for the electrochemiluminescent determination of glucose, lactate and choline

A multi-parametric biochip for glucose, lactate and choline has been developed based on luminol/hydrogen peroxide electrochemiluminescence (ECL). The sensing layers developed were composed of enzyme-bound beads co-entrapped in a photopolymer with luminol-charged beads and spotted at the surface of a glassy carbon electrode (GCE). Lactate oxidase, choline oxidase and luminol were immobilised via electrostatic interactions with DEAE (diethylaminoethyl)-Sepharose whereas glucose oxidase was immobilised, after modification with histidine, by chelation on nickel-charged IDA (imidodiacetic acid)-Sepharose. The luminol immobilisation enabled the achievement of micro-biosensors, present at the surface of the same GCE and free of lateral contamination between each spot by the other’s reaction product. After optimisation of the applied potential, the integration time and the luminol charge in the sensing layer, the electrochemiluminescent H₂O₂ sensor exhibited a detection limit of 2 μM and a working range from 2 μM to 0.5 mM.

The multi-biosensor enabled the concomitant detection of glucose, lactate and choline in the ranges 20 μM–2 mM, 2 μM–0.2 mM and 2 μM–0.2 mM, respectively. Glucose and lactate measurements in complex matrix such as human serum, were in good agreement with the reference methods, without internal calibration of the sample, demonstrating the absence of matrix interference with the present analytical system.     

Effect of biohydrogen by overexpressing small RNA RyhB combined with ldh impairment in novel Klebsiella sp. FSoil 024

Biotechnological hydrogen production is considered as an environmentally sustainable alternative to petrochemical sources or electrolysis. Here, disruption of lactate dehydrogenase (LDH) and metabolic regulation via the small RNA RyhB were adopted to improve hydrogen production in the novel Klebsiella sp. strain FSoil 024. The hydrogen production of FSoil 024-L (Δldh) and FSoil 024-L/R (Δldh/RyhB) from glucose in a 5-L fermenter respectively increased by 40 and 50% compared to the wild type. When glycerol was adopted as a more favorable substrate, FSoil 024-L generated 3.3 L/L of hydrogen after 52 h of fermentation, implying its great potential for the utilization of crude glycerol to produce hydrogen. Overexpression of RyhB downregulated formate biosynthesis in FSoil 024, thereby redirecting NADH toward the hydrogen production pathway. This finding provides new insights into the role of cellular reducing power in hydrogen metabolism and establishes a rationale for improving hydrogen production.     

Application of biosensors in early detection of contamination with lactic acid bacteria during apple juice and concentrate production

The aim of Collective Research Project QUALI-JUICE (COLL-CT-2005, Co Nr 012461) was to introduce the biosensors in control of microbiologically pure apple juice production. Three commercial lactate biosensors were used and compared with enzyme kits assays. Parallel the microbiological analysis of the production process at one juice producing enterprise was done. The results of lactic acid assay with biosensors were in good correlation with those obtained by enzyme kits. The main benefit of biosensor use was shortening of measurement time as compared with assay by enzyme kit and possibility to measure at line. The concentration of l-lactic acid in apple pulp could be correlated with the number of lactic acid bacteria. Pasteurization process was eliminating lactic acid bacteria and the concentration of lactic acid was at this stage not exceeding 0.1 g L⁻¹. The final product (apple concentrate) contained in some cases very high amounts of lactic acid indicating secondary microbiological contamination after pasteurization step. Parallel microbiological analysis of production process and lactate assay indicated that the critical point during production was prolonged vacuum filtration after pasteurization.     

High glycolytic potential does not predict low ultimate pH in pork

Extent of postmortem pH decline influences meat quality development. To better understand physiological determination of ultimate pH (pH_u), we utilized female and castrated male pigs from a line whose selection index includes differentiated pH_u. All genotypes of AMP-activated protein kinase γ3 subunit (AMPKγ3) V199I site were present. The mutant 199II genotype increased pH_u, but only in castrated males. Genotype affected glycolytic potential (GP), but GP was weakly associated with pH_u. A subset of animals was selected based on low (− Gly) and high (+ Gly) residual glycogen content, and compared with AMPKγ3 200Q, which is associated with low pH_u. Both + Gly and 200Q muscle contained glycolytic substrate at 24 h; however, 200Q muscle generated low pH_u and greater lactate compared to + Gly. Additionally,− Gly and + Gly groups exhibited similar pH_u despite a large difference in GP. In conclusion, high GP does not appear to directly impact the extent of postmortem pH decline.     

Photo fermentative hydrogen production using dominant components (acetate, lactate, and butyrate) in dark fermentation effluents

Engineering strategies were applied to promote the phototrophic H₂ production of an indigenous purple nonsulfur bacterium Rhodopseudomonas palustris WP3-5 using major components (i.e., acetate, butyrate, and lactate) of dark fermentation effluents as carbon sources. First, performance of cell growth and photo-H₂ production on each carbon source was examined individually. It appeared that acetate was the most effective carbon source for photo-H₂ production, giving an overall H₂ production rate and H₂ yield of 12.68 ml/h/l and 67.1%, respectively. Next, the effect of substrate concentration of each carbon source on photo-hydrogen production was investigated. Kinetic models were developed to describe the correlation between maximum specific growth rate/specific H₂ production rate and the substrate concentration. The results show that using acetate and lactate as the carbon source, the kinetics for the cell growth and photo-hydrogen production can be described by Monod-type and Michaelis–Menten models, respectively, whereas substrate inhibition occurred when using butyrate as the carbon source. The continuous cultures were also conducted at a hydraulic retention time of 96 h using synthetic dark fermentation soluble metabolites (with a 5 and 10 fold dilution) as the influent. The phototrophic H₂ production efficiency was stably maintained for over 30 days with an overall H₂ yield 10.30 and 11.97 mol H₂/mol sucrose, when using 5-fold and 10-fold diluted dark fermentation effluent, respectively, as the substrate for dark fermentation. This demonstrates the feasibility of using the sequential dark and photo fermentation for high-yield biohydrogen production.