Capillary zone electrophoresis characterization of low molecular weight heparin binding to interleukin 2

A method based on capillary zone electrophoresis (CZE) was used to study the interaction between low molecular weight heparin (LMWH) and interleukin 2 (IL-2). The results showed that the increase of the concentration of LMWH led to the decrease of the peak height and the increase of the peak width of IL-2, but the peak areas were kept constant. The binding constant of IL-2 with LMWH was calculated as 1.2 x 10(6) M(-1) by Scatchard analysis, which is in good agreement with the results found in the references using enzyme-linked immunosorbent assay (ELISA). The results demonstrated that the interaction between IL-2 and LMWH is of fast on-and-off kinetic binding reaction. CZE might be used to study not only slow on-and-off rates interactions, but also fast on-and-off rates ones. The binding constant can be calculated easily, and the method can be applied to study a wide range of heparin-protein interactions. (c) 2005 Elsevier B.V. All rights reserved.

Effects of pressure and molecular weight on the miscibility of polystyrene and cyclohexane

With the aid of Sanchez-Lacombe lattice fluid theory (SLLFT), the phase diagrams were calculated for the system cyclohexane (CH)/polystyrene (PS) with different molecular weights at different pressures. The experimental data is in reasonable agreement with SLLFT calculations. The total Gibbs interaction energy, g*(12) for different molecular weights PS at different pressures was expressed, by means of a universal relationship, as g(12)* =f(12)* + (P - P-0) nu*(12) demixing curves were then calculated at fixed (near critical) compositions of CH and PS systems for different molecular weights. The pressures of optimum miscibility obtained from the Gibbs interaction energy are close to those measured by Wolf and coworkers. Furthermore, a reasonable explanation was given for the earlier observation of Saeki et al., i.e., the phase separation temperatures of the present system increase with the increase of pressure for the low molecular weight of the polymer whereas they decrease for the higher molecular weight polymers. The effects of molecular weight, pressure, temperature and composition on the Flory Huggins interaction parameter can be described by a general equation resulting from fitting the interaction parameters by means of Sanchez-Lacombe lattice fluid theory.

MOLECULAR-WEIGHT DEPENDENCE OF THE GLASS-TRANSITION OF CYCLIC POLYSTYRENE

The glass transition temperature (T(g)) of cyclic polystyrene was measured by differential scanning calorimetry. There was a marked difference in the glass transition behaviour between cyclic and linear polystyrene. In the low molecular weight region (M(n) < 5 x 10(3)), the T(g) of the cyclic polystyrene increased with decreasing M(n), contrary to that of linear polystyrene. With M(n) higher than 5 x 10(3), the T(g) of cyclic polystyrene increased with increasing M(n). The T(g) of cyclic and linear polystyrene approached the same constant value when the M(n) was high enough (M(n) > 10(5)). Combining the results of specific volume, it is believed that the variation of T(g) with molecular weight does not depend only on free volume effects but that configurational entropy is also an important factor.

In vivo growth-inhibition of S 180 tumor by mixture of 5-Fu and low molecular lambda-carrageenan from Chondrus ocellatus

POLYSACCHARIDES; ANTICOAGULANT; SURVIVAL

Antioxidant activity of different molecular weight sulfated polysaccharides from Ulva pertusa Kjellm (Chlorophyta)

Polysaccharides extracted from Ulva pertusa Kjellm ( Chlorophyta) are a group of sulfated heteropolysaccharides, the ulvans. In this study, different molecular weight ulvans were prepared by H2O2 degradation and their antioxidant activities investigated including superoxide and hydroxyl radical scavenging activity, reducing power and metal chelating ability. The molecular weights of natural and degraded ulvans were 151.7, 64.5, 58.0, and 28.2 kDa, respectively, as determined by high performance gel permeation chromatography. Among the four samples, U-3 ( the lowest molecular weight sample) showed significant inhibitory effects on superoxide and hydroxyl radicals with IC50 values of 22.1 mu g mL(-1) and 2.8 mg mL(-1); its reducing power and metal chelating ability were also the strongest among the four samples. All the other samples also demonstrated strong activity against superoxide radicals. The results indicated that molecular weight had a significant effect on the antioxidant activity of ulvan with low molecular weight ulvan having stronger antioxidant activity.

Relevance of molecular weight of chitosan and its derivatives and their antioxidant activities in vitro

The antioxidant potency of different molecular weight (DMW) chitosan and sulfated chitosan derivatives was investigated employing various established in vitro systems, such as superoxide (O-2(.-))/hydroxyl ((OH)-O-.) radicals scavenging, reducing power, iron ion chelating. As expected, we obtained several satisfying results, as follows: Firstly, low molecular weight chitosan had stronger scavenging effect on O-2(.-) and (OH)-O-. than high molecular weight chitosan. For example the O-2(.-) scavenging activity of low molecular weight chitosan (9 kDa) and high molecular weight chitosan (760 kDa) were 85.86 % and 35.50 % at 1.6 mg/mL, respectively. Secondly, comparing with DMW chitosan, DMW sulfated chitosans had the stronger inhibition effect on 0(2)(.-). At 0.05 mg/mL, the scavenging activity on O-2(.-) reached 86.26 %, for low molecular weight chitosan sulfate (9 kDa), but that of low molecular weight chitosan (9 kDa) was 85.86 % at 1.6 mg/mL. As concerning chitosan and sulfated chitosan of the same molecular weight, scavenging activities of sulfated chitosan on superoxide and hydroxyl radicals were more pronounced than that of chitosan. Thirdly, low molecular weight chitosan sulfate had more effective scavenging activity on 02 and (OH)-O-. than that of high molecular weight chitosan sulfate. Fourthly, DMW chitosans and sulfated chitosans were efficient in the reducing power, especially LCTS. Their orders were found to be LCTS > CTS4 > HCTS > CTS3 > CTS2 > CTS1 > CTS. Fifthly, CTS4 showed more considerable ferrous ion-chelating potency than others. Finally, the scavenging rate and reducing power of DMW chitosan and sulfated derivatives increased with their increasing concentration. Moreover, change of DMW sulfated chitosans was the most pronounced within the experimental concentration. However, chelating effect of DMW chitosans were not concentration dependent except for CTS4 and CTS1. (C) 2004 Elsevier Ltd. All rights reserved.

Antihyperlipidemic effects of different molecular weight sulfated polysaccharides from Ulva pertusa (Chlorophyta)

Ulvan, a sulfated polysaccharide from Ulva pertusa, was degraded to yield two low molecular weight fractions U1 and U2. The molecular weights of ulvan and its fractions were determined and varied from 151.6 to 28.2 kDa. They were fed to rats on a hypercholesterolemic diet for 21 days to evaluate and compare the antihyperlipidemic actions. Ulvan-based diet significantly lowered the levels of serum total cholesterol (-45.2%, P < 0.05) and low density lipoprotein cholesterol (LDL-cholesterol, -54.1%, P < 0.05). While U1- and U2-based diets significantly elevated the levels of serum high density lipoprotein cholesterol (HDL-cholesterol, +22.0% for U1, not significant; +61.0% for U2; P < 0.05) and reduced triglyceride (TG, -82.4% for U1, -77.7% for U2; P < 0.05) in rats as compared to control diet. In addition, consumptions of various ulvans significantly increased fecal bile acid excrement. The results indicated that ulvans with different molecular weights exhibited diverse effects on lipid metabolism. The high molecular weight ulvan was effective in serum total and LDL-cholesterol, whereas low molecular weight fractions were in TG and HDL-cholesterol. The fractions were considered to be more beneficial to hyperlipidemia associated with diabetes over ulvan. (C) 2003 Elsevier Ltd. All rights reserved.

The influence of molecular weight of quaternized chitosan on antifungal activity

Quaternized chitosan derivatives with different molecular weights were synthesized in the laboratory. Subsequent experiments were conducted to test their antifungal activities against Botrytis cinerea Pers. (B. cinerea pers.) and Colletotrichum lagenarium (Pass) Ell.et halst (C. lagenarium (Pass) EII.et halst). Our results indicate that quaternized chitosan derivatives have stronger antifungal activities than chitosan. Furthermore, quaternized chitosan derivatives with high molecular weight are shown to have even stronger antifungal activities than those with low molecular weight. (c) 2007 Published by Elsevier Ltd.

Relevance of molecular weight of chitosan-N-2-hydroxypropyl trimethyl ammonium chloride and their antioxidant activities

The antioxidant potency of high/low molecular weight quatemary chitosan derivatives was investigated employing various established systems in vitro, such as superoxide (O-2(center dot-)) and hydroxyl (center dot OH) radicals scavenging, reducing power and iron ion chelating. As expected, we obtained several satisfying results, as follow: firstly, low molecular weight quaternary chitosan had stronger scavenging effect on O-2(center dot-) and center dot OH than high molecular weight quaternary chitosan. Secondly, the reducing power of low molecular weight quaternary chitosan was more pronounced than that of high molecular weight quaternary chitosan. Thirdly, ferrous ion chelating potency were showed to increase first and decrease afterwards with increasing concentration for two kinds of quaternary chitosans, namely, they have not concentration-dependence. However, the scavenging rate and reducing power of high and low molecular weight quaternary chitosans increased with their increasing conc centrations, and hence were concentration-dependent. (c) 2007 Elsevier Masson SAS. All rights reserved.

Different protein tyrosine phosphatase superfamilies resulting from different gene reading frames

Protein tyrosine phosphatases (PTPs) are comprised of two superfamilies, the phosphatase I superfamily containing a single low-molecular-weight PTP (lmwPTP) family and the phosphatase II superfamily including both the higher-molecular-weight PTP (hmwPTP) and the dual-specificity phosphatase (DSP) families. The phosphatase I and H superfamilies are often considered to be the result of convergent evolution. The PTP sequence and structure analyses indicate that lmwPTPs, hmwPTPs, and DSPs share similar structures, functions, and a common signature motif, although they have low sequence identities and a different order of active sites in sequence or a circular permutation. The results of this work suggest that lmwPTPs and hmwPTPs/DSPs are remotely related in evolution. The earliest ancestral gene of PTPs could be from a short fragment containing about 90similar to120 nucleotides or 30similar to40 residues; however, a probable full PTP ancestral gene contained one transcript unit with two lmwPTP genes. All three PTP families may have resulted from a common ancestral gene by a series of duplications, fusions, and circular permutations. The circular permutation in PTPs is caused by a reading frame difference, which is similar to that in DNA methyltransferases. Nevertheless, the evolutionary mechanism of circular permutation in PTP genes seems to be more complicated than that in DNA methyltransferase genes. Both mechanisms in PTPs and DNA methyltransferases can be used to explain how some protein families and superfamilies came to be formed by circular permutations during molecular evolution.

Coagulation removal of melanoidins from biologically treated molasses wastewater using ferric chloride

The pigments (melanoidins) in molasses wastewater are refractory to conventional biological treatment. Ferric chloride was used as coagulant to remove color and chemical oxygen demand (COD) from molasses effluent. Using jar test procedure, main operating conditions such as pH and coagulant dosage were investigated. Under the optimum conditions, up to 86% and 96% of COD and color removal efficiencies were achieved. Residual turbidity in supernatant was less than 5 NTU and Fe3+ concentration was negligible because of effective destabilization and subsequent sedimentation. The results of high performance size exclusion chromatography (HPSEC) show that low molecular weight (MW) fraction of melanoidins is more reactive than high MW fraction and increase in the concentration of the lowest MW organic group is related to the capacity of charge neutralization. Aggregate size measurement reveals the size effect on the settleability of flocs formed, with larger flocs settling more rapidly. Charge neutralization and co-precipitation are proposed as predominant coagulation mechanism under the optimum conditions. (C) 2009 Elsevier B.V. All rights reserved.

Effects of salt stress on carbohydrate metabolism in desert soil alga Microcoleus vaginatus Gom.

The effects of salt stress on carbohydrate metabolism in Microcoleus vaginatus Gom., a cyanobacterium isolated from desert algal crusts, were investigated in the present study. Extracellular total carbohydrates and exopolysaccharides (EPS) in the culture medium produced by M. vaginatus increased significantly during the growth phase and reached a maximum during the stationary phase. The production of extracellular carbohydrates also significantly increased under higher salt concentrations, which was attributed to an increase in low molecular weight carbohydrates. In the presence of NaCl, the production of cellular total carbohydrates decreased and photosynthetic activity was impaired, whereas cellular reducing sugars, water-soluble sugars and sucrose content and sucrose phosphate synthase activity increased, reaching a maximum in the presence of 200 mmol/L NaCl. These parameters were restored to original levels when the algae were transferred to a non-saline medium. Sodium and K+ concentrations of stressed cells decreased significantly and H+-ATPase activity increased after the addition of exogenous sucrose or EPS. The results suggest that EPS and sucrose are synthesized to maintain the cellular osmotic equilibrium between the intra- and extracellular environment, thus protecting algal cells from osmotic damage, which was attributed to the selective exclusion of cellular Na+ and K+ by H+-ATPase.

Degradation of 17 alpha-ethinylestradiol in aqueous solution by ozonation

This study investigates the ozonation of 17 alpha-ethinylestradiol (EE2) in aqueous solution. The affecting factors on the degradation of EE2 were studied and described in details, such as initial EE2 concentration, initial pH value and ozone concentration. In addition, some parameters such as pH. electrical conductivity, mineralization efficiency and degradation products were monitored during the process. The mineralization efficiency of EE2 could reach 53.9%. During the ozonation process the rapid decrease of pH and the sharp increase of electrical conductivity indicated the fort-nation of acidic by-products, small fragments and ions which were confirmed by high performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GUMS) analysis. Results showed that there were intermediate products of smaller molecule with higher polarity produced during the course of EE2 degradation. Then a possible reaction pathway for EE2 degradation involving all intermediates detected is proposed. During the ozonation process EE2 was first oxidized into hydroxyl-semiquinone isomers which were subsequently degraded into low molecular weight compounds such as oxalic acid, malonate, glutarate, and so on. Furthermore. these organic acids are easily oxidized by ozone into carbon dioxide (CO2). This work shows that ozonation process is promising for the removal of EE2. The results can provide some useful information for the potential treatment of EE2 by ozonation in aqueous solution. (c) 2005 Elsevier B.V. All rights reserved.

Dissolution of kaolinite induced by citric, oxalic, and malic acids

Kaolinite is a dominant clay mineral in the soils in tropical and Subtropical regions, and its dissolution has an influence on a variety of soil properties. In this work, kaolinite dissolution induced by three kinds of low-molecular-weight organic acid, i.e., citric, oxalic, and malic acids, was evaluated under far-from-equilibrium conditions. The rates of kaolinite dissolution depended on the kind and concentration of organic acids, with the sequence R-oxalate > R-citrate > R-malate. Chemical calculation showed the change in concentration of organic ligand relative to change in concentration of organic acid in suspensions of kaolinite and organic acid. The effect of organic acid on kaolinite dissolution was modeled by species of organic anionic ligand. For oxalic acid, L-oxalic(2-) and HLoxalic- jointly enhanced the dissolution of kaolinite, but for malic and citric acids, HLmalic- and H2Lcitric- made a higher contribution to the total dissolution rate of kaolinite than L-malic(2-) and L-citric(3-), respectively. For oxalic acid, the proposed model was R-Si = 1.89 x 10(-12) x [(25x)/(1+25x)] + 1.93 x 10(-12) x [(1990x(1))/(1+1990x(1))] (R-2 = 0.9763), where x and x(1) denote the concentrations of HLoxalic and L-oxalic, respectively, and x(1) = 10(-3.81) x x/[H+]. For malic acid, the model was R-Si =4.79 x 10(-12) x [(328-v)/(1+328x)] + 1.67 x 10(-13) x [(1149x(1))/(1+1149x(1))] (R-2 =0.9452), where x and x(1) denote the concentrations of HLmalic and L-malic, respectively, and x(1) = 10(-5.11) x x/[H+], and for citric acid, the model was R-Si = 4.73 x 10(-12) x [(845x)/(1+845x)] +4.68 x 10(-12) x [(2855x(1))/(1+2855x(1))] (R-2 =0.9682), where x and x(1) denote the concentrations of H2Lcitric and L-citric, respectively, and x(1) = 10(-11.16) x x/[H+](2). (c) 2005 Elsevier Inc. All rights reserved.

Total and subcellular distribution of trace elements in the liver of a mother-fetus pair of Dall's porpoises (Phocoenoides dalli)

Total and subcellular hepatic Zn, Cu, Se, Mn, V, Hg, Cd, and Ag were determined in a mother-fetus pair of Dall's porpoises (Phocoenoides dalli). Except for higher fetal Cu concentration, all maternal elements were higher. Elements existed mostly in the cytosol of both animals except in the case of maternal Ag in the microsome and fetal Cu and Ag in the nuclei and mitochondria. In the maternal cytosol, Zn, Mn, Hg, and Ag were present in the high-molecular-weight substances (HMW); Se and V were present in the low-molecular-weight substances (LMW); Cu and Cd were mostly sequestered by metallothionein (MT). In the fetal cytosol, Zn, Se, Mn, Hg, Cd, and Ag were present in the HMW and V in the LMW, while Cu and Ag were mostly associated with MT. MT isoforms were characterized using the HPLC/ICP-MS. Two and four obvious peaks appeared in the maternal and fetal MT fractions, respectively. The highest elemental ion intensities were at a retention time of 7.8 min for the mother, and for the fetus the peak elemental ion intensities occurred at a retention time of 4.3 min, suggesting that different MT isoforms may be involved in elemental accumulation in maternal and fetal hepatocytosols. (C) 2003 Elsevier Ltd. All rights reserved.