Introduction of lysine residues on the light chain constant domain improves the labelling properties of a recombinant Fab fragment

Europium chelates provide a non-radioactive alternative forsensitive labelling of antibodies for diagnostic immunoassays.Lysine residues at antibody surfaces are ready targets for labellingby an isothiocyanate derivative of the europium chelate (Eu³⁺).Here the labelling efficiency of a recombinant anti-human -fetoprotein(hAFP) Fab fragment has been improved by increasing its lysinecontent by protein engineering. Molecular modelling was usedto identify three light chain constant domain surface arginineresidues, R154, R187 and R210, which were mutated to lysineresidues. The mutations did not influence the affinity of thelysine-enriched Fab fragment and its labelling efficiency wasfound to be 40% higher than that of the wildtype Fab fragmentWith low degree of labelling, the affinities of the two Fabfragments were identical and comparable with that of the originalmonoclonal anti-hAFP IgG. With a higher degree of labellingthe affinities of both Fab fragments decreased more than thatof the intact IgG since more lysine residues are available forlabelling in the additional heavy chain constant domains ofthe larger molecule. Electrostatic adsorption and covalent immobilizationof the Fab fragments were characterized by BIAcore^TM and thelysine-enriched Fab fragment was found to be more efficientlyimmobilized to an activated carboxymethyl surface.     

Modelling the structure of latexin-carboxypeptidase A complex based on chemical cross-linking and molecular docking

We have determined the three-dimensional structure of the protein complex between latexin and carboxypeptidase A using a combination of chemical cross-linking, mass spectrometry and molecular docking. The locations of three intermolecular cross-links were identified using mass spectrometry and these constraints were used in combination with a speed-optimised docking algorithm allowing us to evaluate more than 3 x 10(11) possible conformations. While cross-links represent only limited structural constraints, the combination of only three experimental cross-links with very basic molecular docking was sufficient to determine the complex structure. The crystal structure of the complex between latexin and carboxypeptidase A4 determined recently allowed us to assess the success of this structure determination approach. Our structure was shown to be within 4 A r.m.s. deviation of Calpha atoms of the crystal structure. The study demonstrates that cross-linking in combination with mass spectrometry can lead to efficient and accurate structural modelling of protein complexes.     

Treatment of leachate from MSWI bottom ash landfilling with anaerobic sulphate-reducing process

Removal of sulphate and toxic elements from the leachate of a field landfill lysimeter (112m(3)), containing municipal solid waste incineration (MSWI) bottom ash, was studied. The leachate was treated in two parallel laboratory upflow anaerobic sludge blanket (UASB) reactors without and with ethanol as additional carbon source. With ethanol more than 65% of sulphate was removed, while without ethanol removal was negligible. The treatment removed Ba, Ca, Cu, Mn, Mo, Ni, Pb, Tl, Sb, Se, Sr, and Zn of the studied 35 trace and other elements. The sequential extraction of the reactor sludge at the end of runs confirmed that with a few exceptions (Ba, Ca, and Cu) the main mechanism by which the elements were removed was precipitation as sulphides.     

Studies on steam condensation and particle diffusiophoresis in a heat exhanger tube

In this paper, studies on steam condensation and aerosol behaviour in a heat exchanger are presented. The heat exchanger is a model of one single tube of a Passive Containment Condenser (PCC) that is used in the European Simplified Boiling Water Reactor (ESBWR). A hot carrier gas containing nitrogen and steam with Ag and CsOH particles flows through the heat exchanger. The walls of the heat exchanger are cooled with a water jacket, thus causing steam condensation and diffusiophoretic particle deposition. The amount of condensed steam is measured, as well as the temperatures, particle mass concentrations and size distributions before and after the heat exchanger. The experiments are done with different proportions of steam and nitrogen in the carrier gas. Heat and mass transfer in the system are modelled with well known engineering correlations, producing results that agree nicely with the experimental results. The diffusiophoretic particle deposition velocity is shown to be proportional to the steam condensation rate, as expected.     

Effect of surfactant replacement therapy on the outcome of premature infants with respiratory distress syndrome

Lung surfactant replacement has been tested clinically in recent years. In this study the outcome of 31 premature infants with moderate to severe neonatal respiratory distress syndrome (RDS) treated with surfactant was compared to that of 74 prematures with RDS treated conventionally by positive pressure ventilation and supportive care. The groups were well matched for gestational age, birthweight, sex, and Apgar scores at 1 and 5 min. Surfactant treatment resulted in a significant decrease in mortality--from 36.6% in the untreated group to 12.9% in the surfactant-treated group (P < 0.04). This improvement in survival was seen also in prematures with a birthweight < 1,000 g; in the untreated group mortality was 57.6% compared to 23.5% in the treated group (P < 0.05). The incidence of pneumothorax was lower in the treated group--42% vs. 13% (P < 0.01). Surfactant treatment resulted in a trend of more survivors without bronchopulmonary dysplasia or intraventricular hemorrhage, even though surfactant therapy did not change the incidence of either.     

Energy absorption and damage propagation in 2D triaxially braided carbon fiber composites: effects of in situ matrix properties

Results from an experimental program to investigate the propagation of damage and energy dissipation in 2D triaxially braided carbon fiber textile composites (2DTBC) under static conditions are reported. A methodology is presented in which classical concepts from fracture mechanics are generalized to address damage growth in an orthotropic and heterogeneous structural material. Along with results from the experimental program, a novel numerical technique that employs ideas from cohesive zone modeling, and implemented through the use of finite-element analysis, is also presented. The inputs that are required for the discrete cohesive zone model (DCZM) are identified. Compact tension specimen fracture tests and double notched tension tests were carried out to measure the fracture energy (G _Ic), and the maximum cohesive strength (σ _c), of the 2DTBC. The DCZM modeling strategy was independently verified by conducting single edge notched three-point bend tests using a modified three-point bend test fixture. The experimental and numerical analyses were carried out for two different types of 2DTBC made from the same textile architecture but infused with two different resin systems to validate the proposed methodology.     

The use of (green field) biomass pretreatment liquor for fermentative butanol production and the catalytic oxidation of biobutanol

In this study, pretreatment liquor of acid-stored green and yellowish barley silage was used for fermentative acetone–butanol–ethanol (ABE) production. Further, the catalytic oxidation of biobutanol over Pt catalysts was studied to investigate the behaviour of butanol as a fuel in the combustion engine. After the hydrothermal treatment of green and yellowish barley silage followed by enzymatic hydrolysis, approximately 88% and 100% of the available sugars were recovered, respectively. Batch fermentations of pretreatment barley silage liquor, supplemented with gelatinised barley grain, showed good fermentability with total ABE concentrations of 9.0 g/L and 10.9 g/L. Butanol yields of 0.20, 0.17 and ABE yields of 0.28, 0.26 (g/g monosaccharide) were obtained, respectively. In catalytic activity measurements, the conversion of biobutanol became appreciable in the 120–140 °C range, whereas conversions greater than 95% were obtained over 200 °C. Selectivities were also high, although formation of by-products, such as butyraldehyde, was observed.     

Computerized morphometry analysis of epithelial fimbriae nuclear symmetry in BRCA carriers may identify patients at risk for developing ovarian cancer

Serous ovarian tumors may originate in epithelial cells of the fallopian tubes. Computerized morphometry was able to find significant alterations in the fallopian tube epithelium of healthy BRCA carriers. The purpose of this study was to identify a subgroup of BRCA carriers that may be at risk of developing serous ovarian cancer by evaluation of the epithelial nuclear symmetry in the fallopian tubes. Four groups of patients were analyzed; healthy patients, ovarian cancer patients, BRCA carriers, and BRCA noncarriers. All fallopian tubes appeared normal by H&E examination. The ImageProPlus software was used to assess the nuclear symmetry of 65 fimbriae epithelium cells and an artificial neural network algorithm aided in detecting a subpopulation among fimbriae of healthy BRCA carriers at risk for ovarian cancer. Significant differences were found between healthy patients and ovarian cancer patients, and between BRCA carriers and noncarriers. The algorithm was able to accurately predict BRCA carriers with associated ovarian cancer based on fallopian tube nuclear symmetry characteristics. These results reinforce the hypothesis that fimbriae epithelial cells of BRCA carriers may undergo early-stage changes that could predict the risk of progression toward malignancy.     

Studies on mechanical properties of wood fiber reinforced cross-linked starch composites made from enzymatically degraded allylglycidyl ether-modified starch

In a previous work we introduced a new family of thermoset composites of softwood fiber and allylglycidyl ether modified potato starch (AGE-starch with a degree of substitution of 1.3 and 2.3) prepared by hot pressing. To improve the processability of AGE-starch with a DS = 1.3 (LDS-3) and to increase hygromechanical properties, the LDS-3 matrix has now been partially degraded by α-amylase at 45 °C (pH 6) for 0.5, 6 and 18 h. The study shows that already a 30 min enzymatic hydrolysis has a marked effect on the modified starch molecular weight and its thermal properties. The new composites with enzyme hydrolyzed AGE-starch, generically named D-LDS-3, showed good fiber dispersion and excellent interface between the fiber and matrix as studied by SEM. Premixes of D-LDS-3 matrix and fiber showed improved processability. The water vapor absorption was evaluated at 43.2% and 82.2% RH and the stiffness and strength properties were measured. The water uptake was shown to be reduced. The strength of neat matrix D-LDS-3-6 at ambient 68% RH reached 63 MPa and Young’s modulus 3200 MPa and with 40 wt.% wood fiber reinforcement impressive 128 MPa and 4500 MPa, respectively.