Benefit of Interdisciplinary Care in Resource Identification in an Adolescent and Young Adult Oncology Care Model.

There are limited data to identify the best care model to support the vulnerable adolescent and young adult (AYA) oncology population. We sought to compare the impact of AYA physician visits versus interdisciplinary team (IDT) care on AYA-specific resource identification and utilization, as well as to provide a model of AYA oncology care implementation. We identified AYA-aged patients 15-39 years with a current or prior history of cancer seen by the University of Wisconsin Carbone Cancer Center (UWCCC) AYA Oncology Program between January 21, 2021 and May 27, 2021.

A Protein Epitope Targeted by the Antibody Response to Kawasaki Disease.

Background: Kawasaki disease (KD) is the leading cause of childhood acquired heart disease in developed nations and can result in coronary artery aneurysms and death. Clinical and epidemiologic features implicate an infectious cause but specific antigenic targets of the disease are unknown. Peripheral blood plasmablasts are normally highly clonally diverse but the antibodies they encode are approximately 70% antigen-specific 1-2 weeks after infection.

The End of Chimpanzee Research.

In June 2010, Rosie, a descendant of the chimpanzees sent into space, and thirteen others were shipped from New Mexico to a laboratory in Texas for possible use in hepatitis research. They were to be the first group of approximately two hundred chimpanzees to be reintroduced to invasive research. These chimpanzees had been in semiretirement for a decade after being removed from an enormous laboratory that was in egregious violation of the Animal Welfare Act.

Ethical issues in African great ape field studies.

Great apes have been systematically studied in the wild for over half a century. Great apes are now critically endangered and this raises significant ethical issues for field primatologists who study and work to conserve these primates and their habitats. The most immediate ethical concerns involve the well-being of the subjects, but there are also important ethical considerations involved in researchers' interactions with local human populations and extracting industry representatives.

Raising awareness of angina in women.

"If ischaemic heart disease affects women differently from men, then our diagnostic testing, risk stratification schemes and treatment modalities should be sexist as well."

Concise review: scientific and ethical roadblocks to human embryonic stem cell therapy.

Despite the identified therapeutic potential of embryonic stem cells for treating human disease and injury, a number of roadblocks, scientific and ethical, stand in the way of progress toward this goal. We identify six areas of particular interest: tumorigenicity, animal product contamination, genetic compatibility, funding, cell type for transplantation, "embryo-friendly" derivation methods and discuss avenues for moving beyond the difficulties.

Analysis of the binding of the Fab fragment of monoclonal antibody NC10 to influenza virus N9 neuraminidase from tern and whale using the BIAcore biosensor: effect of immobilization level and flow rate on kinetic analysis.

The binding of the Fab fragment of monoclonal antibody NC10 to influenza virus N9 neuraminidase, isolated from tern and whale, was measured using an optical biosensor. Both neuraminidases, homotetramers of 190 kDa, were immobilized to avoid multivalent binding, and the binding of the monovalent NC10 Fab to immobilized neuraminidase was analyzed using the 1:1 Langmuir binding model. A contribution of mass transport to the kinetic constants was demonstrated at higher surface densities and low flow rates, and was minimized at low ligand densities and relatively high flow rates (up to 100 microl/min).

Effects of substitutions in the binding surface of an antibody on antigen affinity.

The interactions between the Fab and single-chain Fv (scFv) fragments of an antibody (NC10) and its antigen, influenza virus neuraminidase, were analysed in the crystal structures of the Fab-neuraminidase and scFv-neuraminidase complexes. To investigate the contribution to binding made by cavities, salt links and hydrogen bonds in the antibody-antigen interface, 14 single amino acid replacements were made at six contact residues in the scFv fragment by site-directed mutagenesis. The binding affinity of each mutant scFv antibody for neuraminidase was determined with a BIAcore optical biosensor.

Nonspecific amine immobilization of ligand can Be a potential source of error in BIAcore binding experiments and may reduce binding affinities.

The interaction of monovalent forms of NC41, an anti-viral neuraminidase antibody, and the antiidiotype antibody 11-1G10 has been used as a model system for BIAcore analysis to demonstrate the potential problems resulting from the nonspecific amine coupling procedure. To avoid complications due to antibody bivalency, monovalent Fab fragments and monomeric recombinant scFvs were used. When immobilized by amine coupling, the 11-1G10 anti-idiotype fragments were found to have an artificially reduced affinity for NC41 compared to the results obtained using site-directed immobilization via C-terminal thiol residue and from solution equilibrium measurements.

Influence of mass transfer and surface ligand heterogeneity on quantitative BIAcore binding data. Analysis of the interaction of NC10 Fab with an anti-idiotype Fab'.

The interaction of monovalent Fab fragments of NC10, an antiviral neuraminidase antibody, and the anti-idiotype antibody 3-2G12 has been used as a model system to demonstrate experimentally the influence of non-ideal binding effects on BIAcore binding data. Because the association rate constant for these two molecules was found to be relatively high (about 5 x 10(5) M-1 S-1), mass transfer was recognised as a potential source of error in the analysis of the interaction kinetics. By manipulation of the flow rate and the surface density of the immobilised ligand, however, the magnitude to this error was minimised.

Identification and minimization of nonideal binding effects in BIAcore analysis: ferritin/anti-ferritin Fab' interaction as a model system.

The interaction of human spleen ferritin with a monoclonal antibody Fab' fragment has been studied as a model system for BIAcore analysis. In particular, the influence of nonideal binding effects has been examined both experimentally and by the theoretical simulation of sensorgram curves. Mass transfer effects were found to have a small but significant influence on the observed binding kinetics of the ferritin/antiferritin Fab' interaction; however, this nonideal behavior could be overcome by systematic manipulation of experimental conditions such as the flow rate and the surface density of the immobilized antigen.

Design and expression of a stable bispecific scFv dimer with affinity for both glycophorin and N9 neuraminidase.

We have designed and produced a stable bispecific scFv dimer (bisFv) by non-covalent association of two hybrid VH-VL pairs derived from an anti-neuraminidase antibody (NC10) and an anti-glycophorin antibody (1C3). The bisFv dimer was demonstrated to have binding activity to the two respective target antigens and was evaluated as a reagent for rapid whole blood agglutination assays. The bisFv was expressed in the periplasm of Escherichia coli, from a secretion vector which comprised two cistrons in tandem under the control of a single lac promoter, inducible with IPTG.

Solution properties of Escherichia coli-expressed VH domain of anti-neuraminidase antibody NC41.

The VH domain of anti-influenza neuraminidase antibody NC41, with and without a C-terminal hydrophilic marker peptide (FLAG), has been expressed in high yield (15-27 mg/L) in Escherichia coli. Both forms were secreted into the periplasm where they formed insoluble aggregates which were solubilized quantitatively with 2 M guanidine hydrochloride and purified to homogeneity by ion-exchange chromatography. The VH-FLAG was composed of three isoforms (pI values of approximately 4.

Recombinant anti-sialidase single-chain variable fragment antibody. Characterization, formation of dimer and higher-molecular-mass multimers and the solution of the crystal structure of the single-chain variable fragment/sialidase complex.

The single-chain antibody variable fragment (scFv), with a 15-residue polypeptide linker (Gly4Ser)3, of monoclonal antibody NC10 was expressed in Escherichia coli and purified to homogeneity. This scFv molecule, refolded from 6 M guanidine hydrochloride, was predominantly a monomer of 27 kDa and was stable on storage at 4 degrees and 20 degrees C. At higher protein concentrations (approximately 5 mg/ml) dimer and higher-molecular-mass multimers were formed and freezing enhanced this aggregation.

Affinity ranking of influenza neuraminidase mutants with monoclonal antibodies using an optical biosensor. Comparison with ELISA and slot blot assays.

A recently developed alternative to the more traditional techniques for studying antigen-antibody interactions has been examined. This method involves the use of an optical biosensor employing surface plasmon resonance detection. In this system one of the reactants is immobilized on the sensor surface and other reactants are passed over the sensor surface sequentially at a constant flow rate.

Determination of relative binding affinity of influenza virus N9 sialidases with the Fab fragment of monoclonal antibody NC41 using biosensor technology.

The relative binding affinities of influenza virus N9 sialidase from term and whale with the Fab fragment of monoclonal antibody NC41 were determined using biosensor technology (Pharmacia BIAcoreTM). The apparent association and dissociation rate constants were measured in real time for the interaction of the Fab with both sialidases, the Fab being immobilised on the sensor surface. Although three-dimensional structural studies have shown that there are no apparent structural differences between the term and whale N9 sialidase epitopes to which the NC41 Fab binds, the apparent binding constant for the interaction with tern N9 sialidase was approximately 2.

Binding affinity of influenza virus N9 neuraminidase with Fab fragments of monoclonal antibodies NC10 and NC41.

Sedimentation equilibrium centrifugation has been applied to determine the affinity and stoichiometry of the interaction between Fab fragments, derived from monoclonal antibodies NC10 and NC41, with influenza virus neuraminidase N9 isolated from either tern or whale. Although the two neuraminidase epitopes recognized by NC10 and NC41 Fab overlap, crystallographic studies have shown that the modes of binding of each Fab are different. The sedimentation equilibrium experiments described here reveal that the binding affinities are also different, with NC10 Fab binding more strongly to each neuraminidase.

Recombinant antineuraminidase single chain antibody: expression, characterization, and crystallization in complex with antigen.

The variable heavy (VH) and variable light (VL) genes of NC10, a monoclonal antibody with specificity toward N9 neuraminidase (NA), were cloned and sequenced. A single chain Fv (scFv) fragment of NC10, consisting of VH and VL domains joined by a peptide linker, was designed, constructed and expressed in the E. coli expression vector pPOW.

Animal welfare and individual characteristics: a conversation against speciesism.

It seems impossible for a human being not to have some point of view concerning nonhuman animal (hereafter animal) welfare. Many people make decisions about how humans are permitted to treat animals using speciesist criteria, basing their decisions on an individual's species membership rather than on that animal's individual characteristics. Although speciesism provides a convenient way for making difficult decisions about who should be used in different types of research, we argue that such decisions should rely on an analysis of individual characteristics and should not be based merely on species membership.

Quantitative analysis of the interaction between lysozyme and monoclonal antibody D1.3.

The method of sedimentation equilibrium has been used to determine the stoichiometry and binding constant for the interaction between hen egg white lysozyme and monoclonal antibody D1.3. The procedures described allow the relative binding affinities of 125I-labelled lysozyme and unlabelled lysozyme to be compared.

Time-dependent ultraviolet absorption changes in proteins at high pH.

The effect of alkali on the ultraviolet absorption of several proteins was examined by difference spectrophotometry. In addition to the well known generation of phenolate ions from tyrosine, time-dependent changes occurred. These were relatively slow in water, but arose more quickly and to a greater degree in 6 M guanidine hydrochloride.

Role of aspartate residues in the cardiac stimulatory activity of anthopleurin-A.

Modification of the polypeptide anthopleurin-A with a carbodiimide and glycine ethyl ester leads to the addition of nearly two Gly to AP-A. Limited amino acid sequence analysis shows that modification occurs principally at Asp-7 and Asp-9. The modified derivative shows no cardiac stimulatory activity.

Conformation and stability of the chymotrypsin inhibitor from winged bean seed (Psophocarpus tetragonolobus (L.) DC).

Spectrophotometric measurement was found to be a sensitive method for evaluating the stability of the chymotrypsin inhibitor from the winged bean. The thermal stability of this protein in aqueous solution was much greater at pH 3 than at pH 8 or pH 11. Evidence from u.