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.     

Epithelial marker genes are expressed in cultured embryonic rat lung and in vivo with similar spatial and temporal patterns

Explants of embryonic lung are often used to characterize lung growth, bronchial tree pattern, and cell differentiation. Most investigators culture lungs for 3-7 days in defined media lacking, e.g., added growth factors or hormones. If growth and differentiation are comparable to that in vivo, these cultures show considerable promise for identifying developmental regulatory molecules and target genes, and for elucidating molecular responses. We used in situ hybridization and RT-PCR to compare times and sites of expression of mRNAs of six epithelial genes in cultured and uncultured fetal rat lungs. These genes, expressed in distal lung of adult rats, are surfactant proteins (SP) A, B, and C; LAR, a receptor-type tyrosine phosphatase; Clara cell secretory protein (CC10, CCSP); and T1alpha. SP-A, SF-B, LAR, and CC10 are expressed by both Clara and Type II cells in adult animals. SP-C and T1alpha are unique markers for Type II and Type I cells, respectively. SP-C, LAR, and T1alpha are expressed before the lung is explanted (Day 13.5); SP-A, -B, and CC10 mRNAs are first detected later. The onset of expression is similar in vivo and in vitro. Although the patterns of expression differ for each mRNA, their sites of expression in culture match those in vivo relative to the bronchial tree. The explanted embryonic lung appears to be an excellent experimental model.     

Determination of total non-sulphonated aromatic amines in tartrazine, sunset yellow FCF and allura red by reduction and derivatization followed by high-performance liquid chromatography.

Free and bound non-sulphonated aromatic amines (NSAA) are determined in the food colours tartrazine, sunset yellow FCF and allura red. After reduction of the bound amines with sodium dithionite, the NSAA are extracted into chloroform, then transferred to aqueous acid solution, diazotized with sodium nitrite and coupled with 2-naphthol-3,6-disulphonic acid, disodium salt (R-salt). Reversed-phase ion-pair liquid chromatography and an absorbance detector at 512 nm are used to analyse the coloured derivatives. Samples of dyes were spiked with known amounts of aniline, 1-naphthylamine, 2- and 4-aminobiphenyl, 4-aminoazobenzene, benzidine, p-cresidine or 4-nitro-p-cresidine bound to R-salt. Recoveries averaged 90% in tartrazine, 65% in sunset yellow FCF and 71% in allura red. Detection limits ranged between 2 and 32 ng/g. A survey of 24 commercial samples revealed levels up to 520 micrograms/g total NSAA. The majority of NSAA are bound to the coupling compound during the manufacturing process and less than 7% remain as free amines in the dye.     

Salt-water up-coning during extraction of fresh water from a tropical island

Rainwater can collect in a lens-shaped region within the rock of a tropical island, and may be separated from the underlying salt water by a sharp interface. This paper presents a nonlinear theory for determining the shape of this interface. The island is assumed to be saturated with rain, and provision is made for the outflow of rain-water through the sides of the island. The effect of a bore well on the shape of the interface is investigated, and the problem is solved using a spectral method. An integral-equation method is also presented for the case when the island has infinite width.     

Implementation of a mechanics-based system for estimating the strength of timber

The most accurate way of determining the strength of lumber requires destructive testing. An intelligent mechanics-based lumber-grading system was developed to provide a better estimation of the strength of a board nondestructively. This system processed X-ray-extracted geometric features (of 1080 boards that eventually underwent destructive strength testing) by using finite element methods to generate associated stress fields. The stress fields were then fed to a feature-extracting-processor, which produced 26 strength predicting features. The best strength predicting features were determined from the coefficient of determination (correlation r/sup 2/) between the features and actual strengths of the boards. The coefficients of determination of each feature (or combination of features), with the actual strength of the board, were calculated and compared. A coefficient of determination of 0.4158 was achieved by using a longitudinal (along the local grain angle) maximum stress concentration (MSC) feature to predict the estimated strength of lumber.     

Neurophysiological Model of the Normal and Abnormal Human Pupil

A model of the pathways controlling the size of the human pupil is presented. Computer simulation of this model demonstrates the role played by each of the elements in the pupil pathways. Simulations of the effects of drugs and a few common abnormalities in the system also help to illustrate the workings of the internal processes. Computer models of this type can be used as teaching aids or as tools for testing of hypotheses regarding the system.     

Self-monitoring processes and Holland Vocational Preferences among college students.

Examined the association between self-monitoring and occupational preferences. 237 undergraduates completed Snyder and Gangestad's (1986) revised self-monitoring scale and Holland's (1977) Vocational Preference Inventory (VPI). Regression analyses showed that men who valued congruence between their beliefs and behavior (i.e., low self-monitors) preferred social occupations. Men who were inclined to modify their behavior to fit social situations (i.e., high self-monitors) preferred enterprising occupations. In contrast, high self-monitoring women preferred artistic occupations. It is argued that these gender-neutral occupations permit high self-monitoring women to use their social skills but without the constraints found in male dominated enterprising occupations. Results support the assumption that people prefer occupations compatible with their social skills. Implications for career counseling and further research are noted. (PsycINFO Database Record (c) 2010 APA, all rights reserved)