Binding of long chain fatty acids to β-lactoglobulin

β-lactoglobulin (BLG), a bovine milk protein that is available commercially in crystalline form, binds long chain free fatty acids (FFA). The binding data were analyzed with a model containing one primary FFA binding site and a large number of weak secondary binding sites. At 37C and pH 7.4, the apparent association constant for binding of FFA to the primary site was of the order of 10⁵ M⁻¹ and that for binding to the secondary sites was approximately 10³ M⁻¹. The strength of binding was: palmitate > stearate > oleate > laurate. The affinity of BLG for palmitate increased as the pH of the incubation medium was raised from 6.5 to 8.7 and decreased as the ionic strength of the medium was raised. Palmitate binding was decreased in the presence of 6 M urea and when the protein either was exposed to elevated temperature or was acetylated prior to incubation. BLG took up methyl palmitate, cetyl alcohol, hexadecane and cholesterol to a lesser extent than FFA. Binding of FFA to BLG was associated with a small increase in the intensity of the fluorescent emission of the protein at 333 mμ. BLG can serve as an FFA acceptor or carrier in biological experiments. FFA released from adipose tissue during in vitro incubation was taken up by BLG. Net transfer of fatty acid to the incubation medium ceased when the molar ratio of FFA to BLG exceeded 1.1.¹⁴C-1-Palmitate bound to BLG was taken up by Ehrlich ascites tumor cells in vitro. At a given palmitate-protein molar ratio, much more labeled fatty acid was taken up by these cells from media containing BLG than from those containing bovine albumin, apparently because FFA is bound less firmly to BLG than to albumin. Special abbreviations used in this text: ν, average molar ratio of bound FFA to total protein; c, molar concentration of FFA in free solution and in equilibrium with that bound to protein; n, number of binding sites in a given class; k’, apparent association constant for binding to a given class of sites.     

Propylene polymerisations with novel heterogeneous combination metallocene catalyst systems

Propylene was polymerised with novel combination metallocene catalyst systems prepared by an emulsion-based heterogenisation method in liquid monomer conditions. The catalyst combinations investigated were rac-dimethylsilanylbis(2-methyl-4-phenyl-1-indenyl)zirconium dichloride/rac-[ethylenebis(2-(tert-butyldimethylsiloxy)indenyl)]zirconium dichloride/methylaluminoxane (MAO) (1 + 2) and rac-dimethylsilanylbis(2-methyl-4-phenyl-1-indenyl)zirconium dichloride/rac-dimethylsilanylbis(2-isopropyl-4-[3,5-dimethylphenyl]indenyl)zirconium dichloride/MAO (1 + 3). The effects of polymerisation temperature and hydrogen on catalyst performance and polymer properties, as well as copolymerisation with hexene and ethylene were investigated. Depending on the polymerisation conditions, M_w of polypropylene varied from 144 to 286 kg/mol for 1 + 2 and from 200 to 390 kg/mol for 1 + 3. Combination 1 + 2 produced broader molecular weight distribution (MWD) than 1 + 3, and a bimodal MWD with clearly separated low- and high-M_w polymer fractions was observed with 1 + 2. The two catalyst systems showed similar hydrogen and hexene responses. Each metallocene precursor showed individual response towards the polymerisation conditions, especially polymerisation temperature, suggesting that interaction between the catalyst active sites was negligible in the studied systems.     

Poisoning effect of CO on ethylene polymerization with Ni(II)-diimine/MAO

CO was not a comonomer but an inhibitor in ethylene polymerization catalyzed over [1,2-bis(2,6-diisopropylphenylimino)]acenaphthene nickel(II) dibromide (1)/MAO. The average number of active sites, , and the average rate constant for chain propagation, of the (1)/MAO system was determined using CO inhibition method based on the assumption that two molecules of CO coordinate to each active center. At 0 °C, the average number of active centers, , was increased with the Al/Ni ratio, and , was not influenced by the Al/Ni ratio. Up to Al/Ni ratio of 3000, the average number of active sites was saturated. Single active site was present at the high ratio of Al/Ni and its reactivity with CO is uniform at 0 °C. The maximum average activity was 5262.07 (kg-PE/mol-Ni/atm/hr) and 65.7% of (1) was converted to form active cation complexes at 0 °C and Al/Ni molar ratio of 5000, while 36.5% of (1) was activated at the Al/Ni molar ratio of 250. Above 30 °C, the complicated CO poisoning behavior was observed because the reactivity and stoichiometry of active centers with CO were not uniform and their thermal stability was very poor.     

Effects of free fatty acids on the binding of steroid hormones to bovine serum albumin

The effects of binding of free fatty acids (FFA) to bovine serum albumin (BSA) on steroid hormone binding to BSA were examined. The FFA studied included myristic, palmitic, stearic, oleic and linoleic acids. The binding coefficient K was estimated from the changes in the equilibrium partition coefficient between the aqueous and the hexane phase caused by the addition to BSA to the aqueous phase. A noticeable effect of FFA binding (molar ratio FFA/BSA, 2∶1) on the affinities of α-estradiol, ethynylestradiol and dehydroisoandrosterone to BSA was not observed: however, the affinities of progesterone, androsterone and testosterone were distinctly enhanced by FFA binding. Furthermore, the elution profiles of gel filtration chromatography clearly showed that progesterone and testosterone are easily liberated from the hormone/BSA complexes and that stronger binding of these hormones to BSA is caused by binding of FFA to BSA. The affinity of ethynylestradiol to BSA is stronger than that of progesterone and testosterone and is not affected by palmitic acid binding to BSA.     

Selective Enrichment of Conjugated Linoleic Acid Isomers in Their Mixtures Using Combined Chemical and Enzymatic Methods

The aim of this study was to selectively enrich t10,c12-conjugated linoleic acid (t10,c12-CLA) and c9,t11-CLA in commercial CLA mixtures using a combination of urea crystallization and lipase-catalyzed esterification. The objective of the urea fractionation is to remove saturated and monounsaturated fatty acids (FA) from the CLA mixtures. CLA-enriched free FA (FFA) mixtures containing 53.8 wt% t10,c12-CLA and 39.1 wt% c9,t11-CLA were produced from the CLA mixtures containing ~34 wt% each of the two CLA isomers by a urea crystallization using methanol and the urea-to-FA weight ratio of 2.5:1. The CLA-enriched FFA mixtures were partially esterified with dodecan-1-ol in a recirculating packed-bed reactor using an immobilized lipase from Candida rugosa to further enrich the t10,c12-CLA and c9,t11-CLA in an FFA fraction and an FA dodecyl ester fraction, respectively, under the optimal conditions, i.e., temperature, 20 °C; FA-to-dodecan-1-ol molar ratio, 1:1; water content, 2 wt% of total substrates; residence time, 5 min; and reaction time, 24 h (for t10,c12-CLA enrichment) and 12 h (for c9,t11-CLA enrichment). After the reaction, an FFA fraction with 72.6 wt% t10,c12-CLA was obtained. Another FFA fraction with 62.0 wt% c9,t11-CLA was recovered after the saponification of the FA dodecyl ester fraction. The yields of t10,c12-CLA and c9,t11-CLA in the FFA fractions were 43.6 and 21.5 wt%, respectively, based on their initial weights in the CLA mixtures.     

Electrochemical study of stable N-alkoxyarylaminyl radicals

The electrochemical study of N-tert-butoxy-2,4-diphenyl-6-tert-butylphenylaminyl (1a), N-tert-butoxy-2,4-bis(4-chlorophenyl)-6-tert-butylphenylaminyl (1b), N-[2-(methoxycarbonyl)-2-propyl]-2,4-diphenyl-6-tert-butylphenylaminyl (2), and N-tert-butoxy-2,4,6-tris(4-chlorophenyl)phenylaminyl radicals (3) was performed by cyclic voltammetry using acetonitrile as the solvent and Bu₄NPF₆ as the supporting electrolyte. On cathodic scan (100 mV/s), all the radicals gave chemically reversible cyclic voltammograms, and the were determined to be −1.405 V (1a), −1.310 V (2a), −1.282 V (2b), and −1.195 V (3) (versus Fc⁺/Fc), respectively. On anodic scan (100 mV/s), on the other hand, 1a, 1b and 2 showed chemically reversible cyclic voltammograms, but 3 exhibited a partially reversible couple even on a scan rate of 500 mV/s, indicating that the cation species of 3 was less stable. The determined for 1a, 1b, 2 and 3 were 0.220, 0.280, 0.318 and 0.294 V (versus Fc⁺/Fc), respectively. The electrochemical data were compared with those of thioaminyl radicals, the corresponding sulfur analogues of 1-3.     

Comparison of Free Fatty Acids Composition of Cuticular Lipids of <Emphasis Type="Italic">Calliphora vicina</Emphasis> Larvae and Pupae

The chemical characterization of the free fatty acid (FFA) fractions of the cuticular lipids of Calliphora vicina larvae and pupae was performed by separating the FFA fraction using high-performance liquid chromatography with laser light scattering detection (HPLC–LLSD) and quantitatively analyzing the FFA using gas chromatography–electron impact mass spectrometry (GC–MS). Thirty-two saturated and unsaturated FFA were identified and quantified in the insect lipids. Cuticular FFA profiles of C. vicina larvae and pupae were compared. Cuticular FFA of larvae and pupae accounted for 70.8 and 77.8 % of the total lipids, respectively. The cuticular lipids of C. vicina larvae contained 24 FFA ranging from 8:0 to 24:0, whereas the cuticular lipids of pupae contained 32 FFA ranging from 6:0 to 26:0. The cuticular lipids of the larvae contained 16 saturated, five monounsaturated, one diunsaturated, and two polyunsaturated FFA. The cuticular lipids of the pupae contained 18 saturated, nine monounsaturated, two diunsaturated, and three polyunsaturated FFA. The major cuticular FFA in C. vicina larvae and pupae was 18:1 (47.6 and 41.7 %, respectively). The highest amounts of total cuticular FFA were detected in larvae of C. vicina (1.7 mg/g of the insect body). The quantities of total cuticular FFA in pupae were smaller (1.4 mg/g of the insect body).     

Optical and electroluminescent properties of polyfluorene copolymers and their blends

Light emitting properties of several polyfluorene (PF) copolymers (P1-P4) and their blends have been investigated. Light emitting diodes were fabricated with the configuration of ITO/PEDOT:PSS/polymer/Ca/Al. The EL peak wavelengths were 421 nm (violet), 505, 513 nm (green) and 570 nm (yellow) for PF copolymers and 510, 535 nm (green) for P1/P2 and P1/P3 blends, respectively. Förster energy transfer in the photoluminescence and electroluminescence of the polymer blends P1/P2 and P1/P3 was studied. The LED using the polymer blend P1/P2 showed a turn-on voltage of 2.5 V and a brightness of 5×10⁴ cd/m² at 7 V. The highest external quantum efficiency was observed to be 3.71% at 5 V. Upon addition of 20 wt% of the green emitter P2 to the violet emitter P1, the device efficiency increased from 1.18 to 3.71%.     

Impacts of Oil Types and Storage Conditions on Milk Fat Quality of Strained Yogurt Immersed in Oil

To maintain strained yogurt freshness, it was shaped into balls (3–3.5 cm diameter), completely immersed in oil, and stored at refrigerated or ambient temperature for 6 months. The impacts of immersing strained yogurt in oil on the topped oil and product's final quality of lipids including fat oxidation, lipolysis, cholesterol oxidation, and conjugated linoleic fatty-acid content (CLNA) (i.e., as freshness indicators), during storage under various storage conditions, were evaluated. Peroxide value (PV), 7-keto-cholesterol, free fatty acids (FFA), and CLNA content were also determined. When stored in light, FFA of strained yogurt immersed in olive or corn oil increased from 0.44 to 1.29 and from 0.12 to 0.58, respectively. Olive oil-topped and corn oil-topped samples exposed to light had 48.9 and 7.9 (meq O₂ kg⁻¹) PV, respectively. Similar trends were reported for strained yogurt fat during storage for up to 6 months. Storage conditions showed little to no effects on the rate of fat and cholesterol oxidation, fat lipolysis, and CLNA content. Strained yogurt freshness indicators were maintained during the extended storage duration without refrigeration. The freshness indicators have a significant impact on enhancing consumer preference of strained yogurt.     

Medium-Chain Enriched Diacylglycerol (MCE-DAG) Oil Decreases Body Fat Mass in Mice by Increasing Lipolysis and Thermogenesis in Adipose Tissue

Medium chain fatty acid (MCFA) escapes the formation of chylomicrons in the small intestine, resulting in energy expenditure through beta-oxidation. Diacylglycerol (DAG) is susceptible to oxidation rather than being stored in the adipose tissue. This study was conducted to verify the effect of MCE-DAG oil on body fat mass in vivo. Male C57BL/6 mice were randomly assigned to four groups (n = 12) as follows: (1) normal diet (18% kcal from fat), (2) canola oil as a control (40% kcal from canola oil), (3) MCE-DAG10 (10% kcal from MCE-DAG + 30% kcal from canola oil), and (4) MCE-DAG20 (20% kcal from MCE-DAG + 20% kcal from canola oil). The body weight and fat mass of MCE-DAG20 group mice were decreased relative to those of control mice (P < 0.05 and P < 0.001, respectively). Serum triacylglycerol (TAG) was decreased in both MCE-DAG10 and MCE-DAG20 groups (P < 0.01 and P < 0.05, respectively). Hormone sensitive lipase (HSL) and adipose triglyceride lipase (ATGL) were increased in the MCE-DAG20 group relative to the control in white adipose tissue (WAT) (P < 0.05). Uncoupling protein 1 (UCP1) was also increased in the MCE-DAG20 group relative to the control in brown adipose tissue (BAT) (P < 0.05). In summary, MCE-DAG reduced body fat mass likely by stimulating lipolysis in WAT and thermogenesis in BAT.     

Synthesis and characterization of alternating fluorene-based copolymers containing diaryl- and non-substituted bithiophene units

A series of soluble alternating fluorene-based copolymers containing diaryl- and non-substituted bithiophene units are synthesized by palladium-catalyzed Suzuki coupling reaction. All polymers demonstrate green colors of photoluminescence (PL) in chloroform, good thermal stability (with decomposition temperatures above 436 °C), and high glass transition temperatures (in the range of 120-144 °C). Owing to the large steric hindrance of diaryl substituents on bithiophenes in the polymers (P2-P4), the aggregation of solids is reduced as well as the solubility is improved, so the performance of their PLED devices are superior to that of the non-substituted polymer (P1). Compared with P1, the introduction of substitutents at 3,3′-position of bithiophene in P2-P4 has significant effects on the photophysical properties of resulting polymers in solution and solid states. Though the PL quantum yield of P1 is much higher than those of diaryl-substituted polymers (P2-P4), the PLED device of P1 has the worst electroluminescence (EL) properties due to the poor solubility of P1. Consequently, among these polymers, the device made of P3 as an emitter has the highest luminance of 2590 cd/m² at 9.5 V. For optimum device performance, a device of P3 blended with PVK can be further enhanced to a brighter luminance of 4284 cd/m² at 18 V.     

Conjugated linoleic acid supplementation in humans: effects on fatty acid and glycerol kinetics

Recent studies with mouse adipocytes have shown that dietary conjugated linoleic acid (CLA) may reduce body fat by increasing lipolysis. The present study examined the effect of CLA supplementation on fatty acid and glycerol kinetics in six healthy, adult women who were participating in a controlled metabolic ward study. These women were fed six CLA capsules per day (3.9 g/d) for 64 d following a baseline period of 30 d. The subjects were confined to a metabolic suite for the entire 94-d study, where diet and activity were controlled and held constant. The rate of appearance (Ra) of glycerol, which indicates lipolytic rates, was similar at baseline and after 4 wk of CLA supplementation at rest (1.87±0.21 and 2.00±0.39 μmol/kg/min, respectively) and during exercise (7.12±0.74 and 6.40±0.99 μmol/kg/min, respectively). Likewise, the Ra of free fatty acids (FFA) was not significantly different after 4 wk of dietary CLA at rest (2.72±0.06 and 2.74±0.12 μmol/kg/min, respectively) or during exercise (6.99±0.40 and 5.88±0.29 μmol/kg/min, respectively). CLA supplementation also had no effect on the percentage of FFA released from lipolysis that were re-esterified. The apparent rate of FFA re-esterification was 65.2±4.2% at rest and 32.1±3.44% during exercise. Four weeks of CLA supplementation had no significant effect on fatty acid or glycerol metabolism in healthy, weight-stable, adult women.     

Electroreduction of lichexanthone

This paper reports the first study on the electrochemical reduction of lichexanthone (1H) (1-hydroxy-3,6-dimethoxy-8-methylxanthen-9-one) on glassy carbon (GC) electrodes in DMSO, using cyclic voltammetry, rotating disc and ring electrodes, and long-term controlled-potential electrolysis. Parameters involving data from cyclic voltammetry and rotating disc electrodes, such as current functions, E_pc1 vs. log ν, E_pc2 vs. log ν, E_pc/2,1 − E_pc1, −I_pc1ox/I_pc1red, I_pc2/I_pc1, E_1/2 vs. log ω, and collection efficiency (rotating disc and ring electrode data), were used to elucidate the reduction mechanism of 1H that involves two one-electron transfers (two reduction peaks in the voltammograms), the first of which, with reversible characteristics, involves electroreduction of 1H, producing a radical anion 1H⁻, whereas the second, with irreversible characteristics, involves electroreduction of 1H⁻, producing a dianion 1H²⁻. Both transfers appear to involve an E_rC_slow-type mechanism with a chemical step consisting of breakage of a bond followed by protonation of residual water, or parent compound, or solvent, etc., to yield 2-hydroxy-4-methoxy-6-methylphenyl 2-hydroxy-4-methoxyphenyl ketone (1H₃), directly, in the case of 1H²⁻ involved. Compound 1H₃ was elucidated by 1D- and 2D-NMR methods. D₀ = 2.66 × 10⁻⁶ cm² s⁻¹ was found for the electrochemical reduction of 1H.     

Synthesis and properties of various poly(diphenylacetylenes) containing tert-amine moieties

The polymerization of diphenylacetylene derivatives possessing tert-amine moieties, such as triphenylamine, N-substituted carbazole and indole, was examined in the presence of TaCl₅-n-Bu₄Sn (1:2) catalyst. A polymer with high molecular weight (M_w = 570 × 10³) was obtained in good yield by the polymerization of diphenylamine-containing monomer 1b, whereas the isopropylphenylamine derivative (1c) gave a polymer with relatively low molecular weight (M_w = 2.4 × 10³). The polymerization of monomer 1d containing cyclohexylphenylamine group did not proceed; however, carbazolyl- and indolyl-containing monomers also produced polymers. Poly(1b), poly(2f) and poly(4b) could be fabricated into free-standing membranes by casting toluene solutions of these polymers. The gas permeability of poly(1b) was too low to be evaluated accurately whereas poly(4b) possessing two chlorine atoms in the repeating unit showed higher gas permeability than that of poly(1b); furthermore, poly(2f) having trimethylsilyl and 3-methylindolyl groups exhibited relatively high gas permeability (). In the cyclic voltammograms of diphenylamino group-containing polymers, poly(1b) and poly(2b), the intensities of oxidation and reduction peaks decreased more than those of carbazolyl-containing poly(2a). The molar absorptivity (?) of poly(1b) at ∼700 nm increased with increasing applied voltage in the UV-vis spectrum.     

Fluorescent coumarin derivatives with large stokes shift, dual emission and solid state luminescent properties: An experimental and theoretical study

Coumarin derivatives containing 8-benzothiazole (C-2) and its difluoroboron bound derivative (C-3) were prepared. Both derivatives show dual emission at 322 nm and 513 nm and large Stokes shift (188 nm), compared to the unsubstituted coumarin (C-1), which shows emission at 356 nm with small Stokes shift of 46 nm. C-2 and C-3 show fluorescence in solid state, in contrast the C-1 is non-fluorescent in the solid state. The excited state intramolecular proton transfer (ESIPT) process of C-2 was fully rationalized by DFT/TDDFT calculations with optimization of the ground state (S₀) and excited state (S₁) geometries. TDDFT calculations propose that the large Stokes shift of C-2 and C-3 are due to the re-distribution of the frontier molecular orbitals at excited states. Study of the potential energy curve of C-2 indicated that the dual emission of the C-2 is due to the simultaneous S₁ and S₃ emission, not the rotamer of the enol form.     

Spectroelectrochemical study of perchloroethylene reduction at copper electrodes in neutral aqueous medium

The potential-dependent chemical reaction of perchloroethylene (PCE) on copper in neutral noncomplexing aqueous media is explored by means of surface-enhanced Raman spectroscopy (SERS), linear sweep voltammetry and preparative electrolysis at controlled potential. Voltammetric peaks associated with copper oxide reduction in Na₂SO₄ solution in the presence and the absence of Cl⁻ are correlated with simultaneously acquired SER spectra. Perchloroethylene undergoes a dechlorination process at potentials at E ≤ −0.3 V vs. Ag/AgCl/KCl (3 M), as shown by the emergence of an intense CuCl stretching band at 290 cm⁻¹ and a CH stretching band together with the presence of Cl⁻ in the catholyte. In the potential region between 0 and −0.9 V vs. Ag/AgCl/KCl (3 M) a broad band assigned to CC structures is observed in the triple-bond region (∼1900 cm⁻¹, FWHM = 180 cm⁻¹). In addition, dichloroethylene (DCE) is detected (but not trichloroethylene (TCE)) in this potential region during preparative electrolysis. At potentials lower than −1 V vs. Ag/AgCl/KCl (3 M) carbon residues are the main product, detected on the copper surface by SERS (and confirmed by XPS), whereas in solution higher levels of dichloroethylene and trichloroethylene are detected with a DCE/TCE ratio below 1.