Disulfide bond assignment in human interleukin-7 by matrix-assisted laser desorption/ionization mass spectroscopy and site-directed cysteine to serine mutational analysis

Interleukin-7 (IL-7) is a proteinaceous biological response modifier that has a bioactive tertiary structure dependent on disulfide bond formation. Disulfide bond assignments in human (h)IL-7 are based upon the results of matrix-assisted laser desorption/ionization (MALDI) mass spectroscopy and Cys to Ser mutational analyses. A gene encoding the hIL-7 was synthesized incorporating Escherichia coli codon usage bias and was used to express biologically active protein as determined by stimulation of precursor B-cell proliferation. MALDI mass spectroscopic analysis of trypsin-digested hIL-7 was performed and compared with the anticipated results of a simulated tryptic digestion. Many of the anticipated hIL-7 tryptic fragments were detected including one with a molecular mass equivalent to the sum of two polypeptides linked through a disulfide bond formed from Cys residues (Cys3 and Cys142). Subsequently, Cys to Ser substitution mutational analyses were performed. A hIL-7 variant with all six Cys substituted with Ser was found to be biologically inactive (EC50 > 1 x 10(-7) M). In contrast, a family of single disulfide bond-forming variants of hIL-7 were constructed by reintroducing Cys pairs (Cys3-Cys142, Cys35-Cys130, and Cys48-Cys93), and each could stimulate cell proliferation with an EC50 of 4 x 10(-9), 2 x 10(-8), and 2 x 10(-9) M, respectively. In single disulfide bond-forming mutants of hIL-7, the ability to stimulate cell proliferation was abolished in the presence of 2 mM dithiothreitol. The results presented strongly suggest that only a single disulfide bond is required for hIL-7 to form a tertiary structure capable of stimulating precursor B-cell proliferation.     

Prenatal care in the inner-city: a cooperative effort

PAHCF and Women and Infants Hospital have been working together for many years to provide high quality, accessible prenatal and obstetrical care to low-income, inner-city residents in the Providence area. Many of these patients have multiple medical, social, and nutritional problems that place them at increased risk for poor pregnancy outcomes. The PAHCF sites offer convenient locations, evening hours, and culturally-sensitive, bilingual staffs; together the five sites provided prenatal care to nearly 1200 patients last year. Women and Infants Ambulatory Care Department provided prenatal care to an additional 1800 women in 1991, including all high risk patients referred for complications or diagnostic evaluations. All patients deliver at Women and Infants, then return to their original source of care for postpartum followup and family planning services. Patient records are readily transferred between the sites, eliminating the need for unnecessary duplication of information or laboratory tests. This close collaboration is mutually beneficial to both the health care providers and the patients whom they serve.     

Chronic interleukin-6 alters NMDA receptor-mediated membrane responses and enhances neurotoxicity in developing CNS neurons

Recent studies show that the cytokine interleukin-6 (IL-6) is expressed at elevated levels in the CNS in several disease states and contributes to the neuropathological process. The mechanisms through which IL-6 exerts its CNS effects are primarily unknown. We have investigated the pathophysiological effects of IL-6 on developing CNS neurons using a culture model system and a chronic treatment paradigm. Here, we show, using current- and voltage-clamp recordings, that chronic IL-6 treatment of developing cerebellar granule neurons increases the membrane and current response to NMDA and that these effects are the primary mechanism through which IL-6 produces an enhanced calcium signal to NMDA. We also show that calcium influx through voltage-sensitive calcium channels contributes to the enhanced calcium signal to NMDA in the IL-6-treated neurons in a developmentally regulated manner and that the membrane depolarization to NMDA is more sensitive to the NMDA receptor antagonist ifenprodil in the IL-6-treated neurons compared with control neurons at a late developmental stage, consistent with a larger proportion of NMDA receptors containing the NMDAR2B subunit in the IL-6-treated neurons. Additional studies show that IL-6 treatment reduces the number of granule neurons in culture and enhances neurotoxicity involving NMDA receptors. These results support a pathological role for IL-6 in the CNS and indicate that NMDA receptor-mediated functions are likely to play a critical role in neuropathological changes observed in CNS diseases associated with elevated CNS levels of IL-6.     

The moderating influence of attitude strength on the susceptibility to context effects in attitude surveys.

In 3 experiments, the authors assessed whether attitude strength moderates the susceptibility of attitudes to item context effects in surveys. In Experiment 1, respondents completed multiple measures of attitude strength. Three weeks later, respondents participated in a context experiment. Results revealed that respondents with weak attitudes exhibited significantly larger context effect for 1 of 2 issues. In Experiment 2, the results of Experiment 1 were conceptually replicated by use of measures of interattitudinal embeddedness to assess attitude strength. In Experiment 3, significant strength-moderated context effects were found when attitude strength was assessed in a multi-item, multidimensional manner but not when it was assessed with a single item. Discussion focuses on measurement and theoretical issues related to the moderation and mediation of context effects in attitude surveys. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

The stress intensity for an edge crack in a semi-infinite orthotropic body

The problem of an edge crack in a semi-infinite plane of linear elastic orthotropic material is studied. The correction factor which relates the stress intensity factor for this problem to that for an isolated crack in an infinite body is evaluated for a range of orthotropic material properties. Calculations are restricted to mode I problems. The method requires the numerical solution of an integral equation, the integrands in which are derived from related complex variable solutions.

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Resumé On étudie le problème d'une fissure de bord dans un plan semi-infini en un matériau linéaire eastique et orthotrope. On évalue pour une gamme de propriétés orthotropes du matériau le facteur de correction qui permet de relier le facteur d'intensité de contraintes relatif à ce problème à celui relatif à une fissure isolée dans un corps infini. Ces calculs sont limités aux problèmes de rupture selon le mode. I. La méthode exige de trouver la solution numérique d'une intégrale dont les intégrants sont tirés de solutions associées à variables complexes.

     

Forces required to jettison a simulated S92 passenger exit: Optimal helicopter underwater egress training techniques

This paper outlines the first known examination of the forces required to jettison a simulated exit used during helicopter underwater egress training (HUET). To capture the forces placed on the simulated cabin exit, a purpose build force plate was designed to replace an existing simulator exit used during HUET. A 25-point map was created to identify specific jettison forces required across the entire exit surface. Ten participants completed a total of 120 underwater egress sequences in–air and in-water from a normal flight and fully compressed crash attenuating seat position. The results indicate that the force required to jettison the simulated exit is significantly different in relationship to location on the exit surface. From the results, it can be concluded that helicopter underwater egress training protocols should ensure that offshore candidates are informed of the different force requirements as well as have the opportunity to practice jettisoning a high physical fidelity exit from a fully compressed crash attenuating seat.     

Modeling the large‐strain constitutive behavior of polycarbonate under isothermal and anisothermal conditions

The tensile behavior of polycarbonate was studied at large strains below the glass‐transition temperature. Experiments were carried out at a series of constant temperatures and also under conditions of falling temperatures. The specimens necked with a natural draw ratio of approximately 2, and the study was mainly focused on the necked material. Isothermal experiments revealed an elastic mechanism that initiated beyond the natural draw ratio. A model consisting of an Eyring process and two Gaussian elastic mechanisms was found to be applicable to both the isothermal and anisothermal stress‐relaxation and stress–strain results. The same model also produced reasonable estimates of the stresses generated during the necking process. In addition, a simple relationship between the isothermal and anisothermal stress relaxation was demonstrated. © 2005 Wiley Periodicals, Inc. J Appl Polym Sci 96: 2105–2116, 2005     

Alpha olefin sulfonates from a commercial SO3-Air reactor

Alpha olefin sulfonate (AOS) can be made by SO₃-air sulfonation of straight chain alpha olefins followed by saponification of the neutralized product. The sulfonation step forms unsaturated sulfonic acids, sultones and sultone sulfonic acids. Hydrolysis of the various sultones yields a mixture of unsaturated and hydroxy sulfonates. Sulfonation of commercial mixtures of straight chain alpha olefins in a large-scale SO₃ falling film unit has given AOS of 1.5–3.0% oil based on active content and tristimulus color of about 40% saturation (2% solution) which is readily bleachable with 1–3% NaOCl to about 10–15% saturation. Performance of AOS made from C₁₅−C₁₈ alpha olefin is comparable to that of the high-foaming C₁₁−C₁₄ LAS in both detergency and dishwashing foam. It is superior to similar products made from internal straight chain olefins. The product shows a low order of toxicity and biodegrad-ability slightly better than that of LAS. A C₁₅−C₁₆ AOS blend is especially attractive in liquid detergent formulations. Presented at the AOCS Meeting, Los Angeles, April 1966.     

Challenges in Liquid‐Phase Exfoliation,Processing, and Assembly of Pristine Graphene

Recent developments in the exfoliation, dispersion, and processing of pristine graphene (i.e., non‐oxidized graphene) are described. General metrics are outlined that can be used to assess the quality and processability of various “graphene” products, as well as metrics that determine the potential for industrial scale‐up. The pristine graphene production process is categorized from a chemical engineering point of view with three key steps: i) pretreatment, ii) exfoliation, and iii) separation. How pristine graphene colloidal stability is distinct from the exfoliation step and is dependent upon graphene interactions with solvents and dispersants are extensively reviewed. Finally, the challenges and opportunities of using pristine graphene as nanofillers in polymer composites, as well as as building blocks for macrostructure assemblies are summarized in the context of large‐scale production.