NMR-STUDY ON THE RELATIONSHIP BETWEEN MISCIBILITY AND ANTIAGING PERFORMANCE OF BLENDS CONTAINING BLOCK-COPOLYMERS

The anti-aging performance of blends of polystyrene (PS), styrene-butadiene triblock copolymers (SBS), and PS/styrene-butadiene (SB)-4A (Carm star SE block copolymer) has been studied by means of C-13 NMR techniques. It is found that the anti-aging performance of these kinds of blends largely depends on their miscibility with PS of different molecular weight M(PS). The larger the quantities of PS solubilized in polybutadiene (PBD) domains, the better the anti-aging performance of the blends. It is also found that the anti-aging performance of these blends has dependence on molecular architectures of the SE block copolymers. For the aged blends, the double bonds of PBD were broken, meanwhile serious cross-linking networks formed in the blends. The proposed anti-aging mechanism is that the PS solubilized in PBD domains can efficiently prevent oxygen molecules from diffusing into PBD domains, therefore, successfully stop the oxidative process of PBD.

CHOLESTEROL SENSOR-BASED ON ELECTRODEPOSITION OF CATALYTIC PALLADIUM PARTICLES

A layer of palladium particles was electrodeposited on a glassy carbon electrode. The dispersed Pd particles resulted in a large decrease in overvoltage for the electrochemical oxidation of H2O2 down to +0.4 V vs. Ag/AgCl, based on which a new kind of cholesterol sensor was fabricated. Cholesterol oxidase was immobilized on the Pd-dispersed electrode by cross-linking with glutaraldehyde and a layer of poly(o-phenylenediamine) (PPD) film was electropolymerized on the enzyme layer. The sensor shows a linear response in the concentration range 0.05-4.50 mmol l-1 with a rapid response of less than 20 s. The polymer film can prevent interference from uric acid and ascorbic acid and also increases the thermal stability of the sensor. The sensor can be used for 200 assays without an obvious decrease in activity.

COBALT PORPHYRIN NAFION FILM ON CARBON MICROARRAY ELECTRODE TO MONITOR OXYGEN FOR ENZYME ANALYSIS OF GLUCOSE

A microcarbon array electrode was modified by the placement of a Nafion film containing cobalt tetramethylpyridyl phorphyrin on its surface. This electrode was applied to the analysis of solution glucose when it was further modified by the immobilization of glucose oxidase on the outermost surface of the Nafion by the cross-linking of serum albumin with glutaraldehyde. The concomitant decrease in the concentration of oxygen, as it was consumed in the enzymatic reaction of glucose with glucose oxidase, was determined by either cyclic voltammetry or a double potential step method at the porphyrin-Nafion catalytic electrode. Glucose could be determined in the range of 0.01-4 mM rapidly, without interference from substances such as ascorbate or other saccharides.

CHARACTERIZATION OF GAMMA-IRRADIATED CRYSTALLINE POLYMER .3. THERMAL-BEHAVIOR OF GAMMA-IRRADIATED POLYAMIDE-1010

Thermal behaviour of gamma-irradiated plain PA1010 and PA1010 containing different amounts of difunctional cross-linking agent BMI was investigated. In DSC endo- and exotherm, it was found that during irradiation, the presence of BMI markedly changes the melting and crystallisation characteristics of PA1010. A supposition that the network of BMI-containing specimens is rather loose in structure was proposed to explain the discrepancy in thermal behaviour between these two kinds of specimens. The supposition was further ascertained by the less brittleness in mechanical property of specimens containing BMI. Besides, the complexity of the thermal behaviour of gamma-irradiated PA1010 was discussed and attributed mainly to the increase in sigma-e, the fold surface free energy of chain fold crystals.

Interaction between phycobilisomes from Porphyridium cruentum and thylakoid membranes from Gymnodinium sp or spinach

Thylakoid membranes were isolated from Gymnodinium sp. and spinach, whereas the phycobilisomes were isolated and purified from red alga Porphyridium cruentum. The absorption spectra of the purified phycobilisomes (PBS) showed three peaks at 548, 564, and 624 nm, respectively, and the ratio of the fluorescence intensity at the lambda(680)(em) to lambda(80)(em5) that at was about 7.3. All these results demonstrated that the purified PBS remained intact. The thylakoid membranes were incubated with the purified phycobilisomes, and the thylakoid membranes, which harbored the phycobilisomes, were purified by sucrose density gradient centrifugation. Meantime, the conjugates of phycobilisome-thylakoid membranes were constructed using glutaraldehyde and further purified. Their characteristics were studied by measuring the absorption spectra and fluorescence emission spectra. The results showed that the phycobilisomes from Porphyridium cruentum can attach to the thylakoid membranes from Gymnodinium sp. and spinach without covalent cross-linking, but the excited energy transfer did not occur. The conjugate of phycobilisome-thylakoid. membranes with covalent cross-linking exhibits the excited energy transfer between the phycobilisomes and the thylakoid membranes.

A transglutaminase from Chinese shrimp (Fenneropenaeus chinensis), full-length cDNA cloning, tissue localization and expression profile after challenge

Transglutaminase can catalyze the cross-linking reaction between soluble clotting protein molecules from the plasma for prevention of excess blood loss from a wound and obstructing micro-organisms from invading the wound in crustaceans. A novel transglutaminase (FcTG) gene was cloned from hemocytes of Chinese shrimp Fenneropenaeus chinensis by 3' and 5' rapid amplification of cDNA ends (RACE) PCR. The full-length cDNA consists of 2972 bp, encoding 757 amino acids with a calculated molecular mass of 84.96 kDa and a theoretical isoelectric point of 5.61. FcTG contains a typical transglutaminase-like homologue (TGc domain: E-value = 1.94e-38). Three catalytic sites (Cys-324, His-391 and Asp-414) are present in this domain. The deduced amino acid sequence of FcTG showed high identity with black tiger shrimp TG, kuruma shrimp TG and crayfish TG. Transcripts of FcTG mRNA were mainly detected in gill, lymphoid organ and hemocytes by RT-PCR. RNA in situ hybridization further confirmed that FcTG was constitutively expressed in hemocytes both in the circulatory system and lymphoid organ. The variation of mRNA transcription level in hemocytes and lymphoid organ following injection of killed bacteria or infection with white spot syndrome virus (WSSV) was quantified by RT-PCR. The up-regulated expression of FcTG in shrimp lymphoid organ following injection of bacteria indicates that it is inducible and might be associated with bacterial challenge. (c) 2006 Elsevier Ltd. All rights reserved.

Characterizing a monotopic membrane enzyme. Biochemical, enzymatic and crystallization studies on Aquifex aeolicus sulfide:quinone oxidoreductase

Monotopic membrane proteins are membrane proteins that interact with only one leaflet of the lipid bilayer and do not possess transmembrane spanning segments. They are endowed with important physiological functions but until now only few of them have been studied. Here we present a detailed biochemical, enzymatic and crystallographic characterization of the monotopic membrane protein sulfide:quinone oxidoreductase. Sulfide:quinone oxidoreductase is a ubiquitous enzyme involved in sulfide detoxification, in sulfide-dependent respiration and photosynthesis, and in heavy metal tolerance. It may also play a crucial role in mammals, including humans, because sulfide acts as a neurotransmitter in these organisms. We isolated and purified sulfide:quinone oxidoreductase from the native membranes of the hyperthermophilic bacterium Aquifex aeolicus. We studied the pure and solubilized enzyme by denaturing and non-denaturing polyacrylamide electrophoresis, size-exclusion chromatography, cross-linking, analytical ultracentrifugation, visible and ultraviolet spectroscopy, mass spectrometry and electron microscopy. Additionally, we report the characterization of its enzymatic activity before and after crystallization. Finally, we discuss the crystallization of sulfide:quinone oxidoreductase in respect to its membrane topology and we propose a classification of monotopic membrane protein crystal lattices. Our data support and complement an earlier description of the three-dimensional structure of A. aeolicus sulfide:quinone oxidoreductase (M. Marcia, U. Ermler, G. Peng, H. Michel, Proc Natl Acad Sci USA, 106 (2009) 9625-9630) and may serve as a reference for further studies on monotopic membrane proteins. (C) 2010 Elsevier B.V. All rights reserved.

Improved procedure for preparation of covalently bonded cellulose tris-phenylcarbamate chiral stationary phases

The classical method for preparation of covalently boned cellulose derivative chiral stationary phases (CSP) with diisocyanate as spacer was improved. Diisocyanate was firstly allowed to react with 3-aminopropyltriethoxysilane, and the resulting product was then applied as the spacer reagent to immobilize cellulose derivatives onto silica gel. Influences of the amount and the length of the spacer on the optical resolution ability of the CSP were investigated. Comparing improved procedure to classical diisocyanate method, the cross-linking between the glucose units of the cellulose derivatives was avoided to the most extent. With the improved procedure, regio-nonselective ways could be adopted to prepare covalently bonded CSP, which showed an advantage for the rapid preparation.