RP-HPLC binding domains of proteins

Procedures have been developed to identify the chromatographic binding domains of horse heart cytochrome c (Cyt c) and bovine growth hormone (bGH) during their interaction with reversed-phase sorbent materials. The procedure involves adsorption of the protein solute to the chromatographic sorbent, followed by proteolytic cleavage. Comparison of the proteolytic map obtained for Cyt c and bGH in free solution with the corresponding map obtained when these proteins are adsorbed to the chromatographic sorbent revealed significant differences in the digestion pattern. Following characterization of the peptides generated in both maps, the results indicated that specific regions on the surface of both Cyt c and bGH are inaccessible to tryptic cleavage when adsorbed to the hydrophobic surface of both a C-4 and a C-18 sorbent. Based on the assumption that the region of the protein surface that is in contact with the sorbent remains intact and bound to the sorbent during the digestion step, while the protein surface that is exposed to the solvent is accessible to proteolysis, the regions that were inaccessible to tryptic digestion were found to correspond to hydrophobic domains on the protein surface. These results also suggest that the three-dimensional structures of these proteins remain largely intact upon adsorption to the hydrophobic surface.     

Systemic lupus erythematosus in three ethnic groups: II. Features predictive of disease activity early in its course. LUMINA Study Group. Lupus in minority populations, nature versus nurture

OBJECTIVE: To determine the factors associated with disease activity in patients with recent-onset (< or =5 years) systemic lupus erythematosus (SLE) who were of Hispanic, African-American, or Caucasian ethnicity. METHODS: Incident and prevalent cases of SLE, as defined by the American College of Rheumatology criteria for SLE, among the 3 ethnic groups were identified in Alabama (The University of Alabama at Birmingham) and Texas (The University of Texas-Houston Health Science Center and The University of Texas Medical Branch at Galveston). Variables from the sociodemographic, clinical, immunologic, immunogenetic, behavioral, and psychological domains were obtained using validated instruments. Disease activity was ascertained with the Systemic Lupus Activity Measure (SLAM). Stepwise domain regressions with SLAM score as the dependent variable were performed. Final ethnic-specific and overall regression models were obtained by entering variables that were retained in the domain regressions. RESULTS: SLAM scores at study entry were higher in the African Americans (mean +/- SD 12.6 +/- 6.9) and Hispanics (11.0 +/- 6.2) than in the Caucasians (8.5 +/- 3.7) (P < or = 0.001). The final overall regression model (R2 = 28%) for higher SLAM score included the following variables: African-American ethnicity, lack of private health insurance, abrupt disease onset, presence of anti-Ro antibodies, absence of HLA-DRB1*0301, higher levels of helplessness, and abnormal illness-related behaviors. CONCLUSION: Socioeconomic, immunologic, immunogenetic, behavioral, and psychological variables were all predictive of disease activity early in the course of SLE, irrespective of ethnic group. However, there remain ethnic group differences in disease activity that were not explained by these factors.     

Potassium channels: a computer prediction of structure and selectivity

Model structures for the pore of the potassium channels Shakerand ROMK1 are predicted. The models arise from computer simulationsand suggest reasons for the striking selectivity of these channelsfor K+ and the blocking of ROMK1 by internal Mg²⁺. The modelledstructure of the Shaker pore is supported by mutagenesis data.The mutagenesis experiments indicate the side chains responsiblefor binding to blocking agents [tetraethylammonium (TEA) andcharybdotoxin (CTX)] and the model has these side chains suitablyoriented for binding. An aromatic K+ binding site part way downthe pore is also predicted by the Shaker pore model.     

Evaluation of oxidative stability of canola oils by headspace analysis

Lipid oxidation is a major factor affecting flavor quality and shelf life of vegetable oils. Oxidative stability is therefore an important criterion by which oils are judged for usefulness in various food applications. In this study a method based on headspace analysis was developed to evaluate relative oxidative stability of canola oils. The method does not require the use of chemicals, involves minimal sample preparation, and can be performed on a relatively small sample size in comparison with traditional wet chemical methods. Canola oils freshly extracted in the laboratory from different seed samples were subjected to accelerated oxidation and analyzed for PV by standard methods and headspace volatiles by solid phase microextraction/GC-MS. Forward stepwise regression analysis of the data revealed a relationship between PV and headspace concentration of the volatile lipid oxidation products hexanal and trans,trans-2,4-heptadienal. The PV calculated using this formula correlated (R ²=0.73) with those measured by conventional methods. Presented in part at the 96th Annual Meeting of the AOCS, 1–4 May 2005, Salt Lake City, UT.     

Pheromonal secretions from glands on the 5th abdominal sternite of hydropsychid and rhyacophilid caddisflies (Trichoptera)

Extracts of different body parts of adult Trichoptera were tested for electrophysiological activity. Extracts of the IVth and Vth abdominal sternites of femaleHydropsyche angustipennis, Rhyacophila nubila, andR. fasciata, containing a paired exocrine gland, elicited significant electroan-tennographic responses when tested on conspecific male antennae. The paired gland occurs also in males of all the species, and inH. angustipennis, extracts from males were more active than female extracts when tested on male antennae. Female and male extracts from all species were analyzed by gas chromatography with simultaneous flame ionization and electroantennographic detection (EAD). EAD-active peaks in female extracts, stimulating male antennae, were identified inH. angustipennis as nonan-2-one; and inR. nubila andR. fasciata as heptan-2-one, heptan-2-ol, nonan-2-one, and nonan-2-ol. EAD-active components from maleH. angustipennis stimulating male antennae were octan-2-one, nonan-2-one (major peak), (Z)-6-nonen-2-one, decan-2-one, and a methylbranched decan-2-one. Female extracts and synthetic mixtures of compounds identified from femaleH. angustipennis andR. fasciata were tested for attractivity in the field. High catches with control traps obscured the results, but a synthetic mixture of the four identified compounds was significantly attractive and not different from female extracts for attracting maleR. fasciata. InH. angustipennis, a synthetic six-component male blend, in which nonan-2-one was the major component, attracted significant numbers of male and femaleH. angustipennis. Extracts of maleR. nubila andR. fasciata contained acetophenone and hexanoic and octanoic acids but did not have any electrophysiological or behavioral activity on either male or female antennae of conspecifics. The occurrence of a female sex pheromone inRhyacophila and an aggregation pheromone inHydropsyche corresponds to earlier described differences in mating behaviors in the Rhyacophilidae and Hydropsychidae.     

Structure–property relationships for sulfonated poly(butylene terephthalate)

Structure–property relationships have been developed for sulfonated poly(butylene terephthalate) copolymers. The compositional variables investigated were sulfonate content, molecular weight, and polymer endgroup composition, and the fundamental polymer properties evaluated were melt viscosity, crystallization kinetics, and impact strength. It was found that all compositional variables significantly affect all of the polymer properties of interest. The most interesting effect is the influence of polymer endgroup composition on polymer properties. The trends indicate that the carboxylic acid endgroups form intermolecular interactions with sodium sulfonate groups, resulting in a decrease in the strength of intermolecular ionic interactions between sodium sulfonate groups. © 2006 Wiley Periodicals, Inc. J Appl Polym Sci 100: 4662–4771, 2006     

Direct Observation of Triplet Reactions of Anthraquinone Vat Dyes Using Nanosecond Laser Photolysis

The triplet-triplet spectra of three commercial anthraquinone vat dyes (C.I. 67300, C.I. 59100 and C.I. 60515) were studied in solution at room temperature using laser photolysis. The triplet states of these dyes react with oxygen, and the rate constants for the quenching process were measured, together with the rates of energy transfer from the triplet state to anthracene. These observations are considered in relation to the photoreactivity of these dyes.     

Characterization of a nylon 6/poly(methyl acrylate) copolymer produced by radiation grafting

Acrylic acid has been grafted onto nylon 6 by the mutual γ-irradiation technique. Methods are described for removing poly(acrylic acid) homopolymer and ungrfted nylon, the latter involving intermediate conversion to the calcium ionomer. The pure graft copolymer in its methylated form, viz., nylon 6/poly(methyl acrylate), NY/PMA, was characterized by light scattering in mixed solvents to yield the true molecular weight (3.2 × 10⁶) as well as the molecular weights M_PMA(domain) and M_NY(domain) of the PMA and NY portions, respectively. The molecular weight M_PMA of the grafts was measured after hydrolysis of the backbone, and the molecular weight M_NY of the backbone was determined via a previously devised indirect procedure. Comparisons of M_PMA(domain) with M_PMA and of M_NY(domain) with M_NY gave ? 7 nylon chains and ? 17 poly(methyl acrylate) chains per copolymer molecule. Chain transfer and bimolecular terminatin during grafting are proposed as probable contributory factors to the branched structure of the copolymer.     

Scaling laws and polystyrene networks: a quasi-elastic light scattering study

Randomly crosslinked networks with a range of crosslink densities were prepared by γ-irradiation. Quasielastic light scattering measurements were made on the gels swollen to equilibrium in cyclohexane at 308, 318 and 333 K. The longitudinal osmotic modulus M_os was obtained from the intensity of the dynamically scattered light, and the cooperative diffusion coefficient D_c calculated from the relaxation rate of the autocorrelation function. The concentration dependence of these parameters at the theta temperature was found to be consistent with the scaling predictions, but at higher temperatures where excluded volume conditions prevail, deviations from scaling behaviour were observed. These discrepancies are probably caused by defects in the network structure.