Preferential exclusion of sucrose from recombinant interleukin-1 receptor antagonist: role in restricted conformational mobility and compaction of native state

Understanding the mechanism for sucrose-induced protein stabilization is important in many diverse fields, ranging from biochemistry and environmental physiology to pharmaceutical science. Timasheff and Lee [Lee, J. C. & Timasheff, S. N. (1981) J. Biol. Chem. 256, 7193-7201] have established that thermodynamic stabilization of proteins by sucrose is due to preferential exclusion of the sugar from the protein's surface, which increases protein chemical potential. The current study measures the preferential exclusion of 1 M sucrose from a protein drug, recombinant interleukin 1 receptor antagonist (rhIL-1ra). It is proposed that the degree of preferential exclusion and increase in chemical potential are directly proportional to the protein surface area and that, hence, the system will favor the protein state with the smallest surface area. This mechanism explains the observed sucrose-induced restriction of rhIL-1ra conformational fluctuations, which were studied by hydrogen-deuterium exchange and cysteine reactivity measurements. Furthermore, infrared spectroscopy of rhlL-1ra suggested that a more ordered native conformation is induced by sucrose. Electron paramagnetic resonance spectroscopy demonstrated that in the presence of sucrose, spin-labeled cysteine 116 becomes more buried in the protein's interior and that the hydrodynamic diameter of the protein is reduced. The preferential exclusion of sucrose from the protein and the resulting shift in the equilibrium between protein states toward the most compact conformation account for sucrose-induced effects on rhIL-1ra.     

Situating natural language understanding within experience-based design

Building useful systems with an ability to understand "real" natural language input has long been an elusive goal for Artificial Intelligence. Well-known problems such as ambiguity, indirectness, and incompleteness of natural language inputs have thwarted efforts to build natural language interfaces to intelligent systems. In this article, we report on our work on a model of understanding natural language design specifications of physical devices such as simple electrical circuits. Our system, called KA, solves the classical problems of ambiguity, incompleteness and indirectness by exploiting the knowledge and problem-solving processes in the situation of designing simple physical devices. In addition, KA acquires its knowledge structures (apart from a basic ontology of devices) from the results of its problem-solving processes. Thus, KA can be bootstrapped to understand design specifications and user feedback about new devices using the knowledge structures it acquired from similar devices designed previously.In this paper, we report on three investigations in the KA project. Our first investigation demonstrates that KA can resolve ambiguities in design specifications as well as infer unarticulated requirements using the ontology, the knowledge structures, and the problem-solving processes provided by its design situation. The second investigation shows that KA's problem-solving capabilities help ascertain the relevance of indirect design specifications, and identify unspecified relations between detailed requirements. The third investigation demonstrates the extensibility of KA's theory of natural language understanding by showing that KA can interpret user feedback as well as design requirements. Our results demonstrate that situating language understanding in problem solving, such as device design in KA, provides effective solutions to unresolved problems in natural language processing.     

Cell-mediated immunological responses in cervical and vaginal cancer patients immunized with a lipidated epitope of human papillomavirus type 16 E7

Human papillomavirus (HPV) infection has been causally associated with cervical cancer. We tested the effectiveness of an HLA-A*0201-restricted, HPV-16 E7 lipopeptide vaccine in eliciting cellular immune responses in vivo in women with refractory cervical cancer. In a nonrandomized Phase I clinical trial, 12 women expressing the HLA-A2 allele with refractory cervical or vaginal cancer were vaccinated with four E786-93 lipopeptide inoculations at 3-week intervals. HLA-A2 subtyping was also performed, and HPV typing was assessed on tumor specimens. Induction of epitope-specific CD8+ T-lymphocyte (CTL) responses was analyzed using peripheral blood leukapheresis specimens obtained before and after vaccination. CTL specificity was measured by IFN-gamma release assay using HLA-A*0201 matched target cells. Clinical responses were assessed by physical examination and radiographic images. All HLA-A*0201 patients were able to mount a cellular immune response to a control peptide. E786-93-specific CTLs were elicited in 4 of 10 evaluable HLA-A*0201 subjects before vaccination, 5 of 7 evaluable HLA-A*0201 patients after two vaccinations, and 2 of 3 evaluable HLA-A*0201 cultures after all four inoculations. Two of three evaluable patients' CTLs converted from unreactive to reactive after administration of all four inoculations. There were no clinical responses or treatment toxicities. The ability to generate specific cellular immune responses is retained in patients with advanced cervical cancer. Vaccination with a lipidated HPV peptide epitope appears capable of safely augmenting CTL reactivity. Although enhancements of cellular immune responses are needed to achieve therapeutic utility in advanced cervical cancer, this approach might prove useful in treating preinvasive disease.     

Power-dependent dispersion in far-off-resonant raman scattering

We investigate the saturation effects of power broadening, Stark shifting, and population transfer on Stokes conversion in stimulated Raman scattering. We do not make the usual rotating wave approximation because the detuning from the next electronic state is assumed to be in the optical regime. Retaining the counter-rotating terms allows an exact determination of the pump and Stokes indexes of refraction. Steady-state solutions for the Stokes intensity and phase are obtained and the effects of making the rotating wave approximation (RWA) are discussed. Finally, we examine the behavior of these solutions for Stokes conversion in hydrogen gas when geometric propagation is appropriate.     

Integrating trust into the CyberCraft Initiative via the Trust Vectors model [Application Notes]

This research supports the hypothesis that the Trust Vector model can be modified to fit the CyberCraft Initiative, and that there are limits to the utility of historical data. This research proposed some modifications and expansions to the Trust Model Vector, and identified areas for future research.