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Promotion of monolayer formation and high expression of ammonia metabolism of primary rat hepatocytes on arginine-glycine-aspartic acid-containing peptide-coated polystyrene dish.

Cell adhesive peptide Arg-Gly-Asp (RGD) was immobilized using ProNectin F (PnF) on a nontreated polystyrene petri dish at a PnF density of 20 ng cm(-2), which is sufficient for primary rat hepatocyte immobilization. The density of PnF on the dish affects cell morphology and expression of the differentiated functions within the range of 2-2500 ng cm(-2). An optimal monolayer state with defined cell boundaries and hepatocyte nuclei was formed on a 2500 ng cm(-2) PnF-coated petri dish, and the ammonia metabolic function was expressed at as high a level as in the hepatocyte/spheroid. We conclude that 2500 ng cm(-2) PnF enhances the morphological stability and expression of liver-specific functions of the hepatocyte.

Ammonia↗

Exploring time-dependent structural changes during the cold crystallization process of isotactic polystyrene by infrared spectroscopy and multivariate curve resolution.

The present study attempts an application of Fourier transform infrared (FT-IR) spectroscopy in conjunction with multivariate curve resolution (MCR) techniques to explore the structural evolution of isotactic polystyrene (iPS) during the cold crystallization process. The focus of the present study is placed on the performance of MCR techniques, e.g., orthogonal projection (OP), alternating least squares (ALS), and fixed-size moving window evolving factor analysis (FSMWEFA), and the interpretability of spectral changes in the investigated chemical process. As a result, valuable information and conclusions about the structural evolution of iPS during the crystallization process can be extracted: when the amorphous phase of iPS changes, the ordering of the phenyl rings takes place first, and then the polymer chains adjust their local conformations to form short 3(1) helix structures. Furthermore, according to intensity profiles of the spectral variations, the ordering of the phenyl rings proceeds more intensely than the formation of ordered local chains, and the structural evolution of iPS occurs even during the induction period. The spectral variations resulting from the conformational changes in the 3(1) helical structures depend on the sequence length of the helical chains: the longer the polymer chain is, the smaller the corresponding band variations are. It has been demonstrated that the combination of FTIR spectroscopy and chemometric MCR techniques is very promising for the analysis of the crystallization process of polymers. MCR is a powerful tool for analyzing and visualizing spectral data and integrating them with other information, making spectral intensity variations more amenable to interpretation in order to explore the molecular dynamics of polymers.

Algorithms↗

Production of L-DOPA by tyrosinase immobilized on modified polystyrene.

Mushroom tyrosinase was immobilized on modified polystyrene- polyamino styrene (PSNH) and polymethylchloride styrene (PSCL)-to produce L-DOPA from L-tyrosine. Glutaraldehyde was used as an activating agent for the PSNH to immobilize the tyrosinase, and 10% (w/v) glutaraldehyde was optimal in conferring the highest specific activity (11.96 U/g) to the PSNH. Methylchloride on the PSCL was directly linked with the tyrosinase, and 1.5 mmol of Cl/g was optimal in attaining the specific activity of 17.0 U/g. The temperature and optimal acidity were, respectively, 60 degrees C and pH 5.5 for the PSNH, and 70 degrees C and pH 3.0 for the PSCL. In a 50-mL batch reactor working over 36 h, the L-DOPA production rate at 30 degrees C was 1.44 mg/(L x h) for the PSNH and 2.33 mg/(L x h) for the PSCL. The production rate over 36 h was 3.86 mg/(L x h) for the PSNH at 60 degrees C and 5.54 mg/(L x h) for the PSCL at 70 degrees C. Both of the immobilized enzymes showed a remarkable stability with almost no change in activity after being stored wet. The operational stability study indicated a 22.4% reduction in L-DOPA production for the PSNH and an 8.63% reduction for the PSCL over seven runs (each run was for 144 h at 30 degrees C) when the immobilized enzymes were used under turnover conditions. The immobilized tyrosinase was more stable on the PSCL than on the PSNH.

Agaricales↗

N-terminal fibronectin 30-kDa fragment mediates the immobilization of soluble fibrin by factor XIIIa-coated polystyrene beads.

Polystyrene beads coated with thrombin-activated Factor XIII (plasma transglutaminase, plasma transamidase) retained soluble 125I-fibrin, if a 30-kDa fragment of fibronectin was present. The fragment was obtained by proteolytic cleavage of plasma fibronectin with trypsin, but was also available with plasmin or thrombin. It represented a fibrin-binding domain at the N-terminus of each of the two subunit chains and contained close to its amino end a transamidase-reactive site. Fibronectin was unable to mediate the binding of 125I-fibrin to Factor XIIIa-coated beads. 125I-fibrinogen was hardly recognized by the beads even in presence of the fibronectin fragment. The relatively slow binding of 125I-fibrin was inhibited by 0.15 mM putrescine or by a pretreatment of the coated beads with EDTA or N-ethylmaleinimide indicating the involvement of a transamidation in the binding reaction. Immobilization of 125I-fibrin in presence of the fibronectin fragment is assumed to require a covalent cross-linking of the two ligands at the immobilized transamidase giving rise to a product which is retained strongly. The possibility is discussed that a surface-attached transamidase might act as a fibrin receptor which requires the fibronectin fragment as a cofactor.

Edetic Acid↗

Continuous flow peptide synthesis on aminopolyoxyethylene-polystyrene graft copolymer using Fmoc-strategy.

An insoluble graft copolymer consisting of the covalently bound polyoxyethylene to cross-linked polystyrene (HO-POE-PS) was prepared by anionic polymerization of ethylene oxide on the resin. The copolymer was then converted to the corresponding amino-polymer (H2N-POE-PS) and the latter was employed as the solid carrier for peptide synthesis. Although HO-POE-PS has successfully been employed as a carrier for peptide synthesis by the standard shaking procedure using t-butoxycarbonyl-amino acids, now we deemed it of interest to test its suitability for the continuous flow synthesis. Thus, the C-terminal octapeptide of the porcine insulin B chain (B23-30) was prepared by this procedure using a photolabile anchoring group and fluoren-9-ylmethoxycarbonyl-amino acids. All the reactions were carried out in a continuous flow manner in a steel column under pressure using a high-performance liquid chromatography (HPLC) system. At the end of the synthesis, a sample of the protected peptide was cleaved from the support by photolysis. For the cleavage of another sample, an aqueous solution of sodium carbonate was employed. The protected peptide was purified on silica gel and Sephadex-LH 20. All the protecting groups of a sample of the octapeptide were removed with piperidine/dimethylformamide and trifluoroacetic acid and the deblocked peptide was purified by ion-exchange chromatography. The free peptide was shown to be homogeneous by thin-layer chromatography, HPLC, and electrophoresis. The identify of the free octapeptide was confirmed by amino-acid analysis, 13C-nuclear magnetic resonance measurement and field-desorption mass spectrometry. The peptide was also shown to be free of racemization.

Chromatography, High Pressure Liquid↗

I-domain of lymphocyte function-associated antigen-1 mediates rolling of polystyrene particles on ICAM-1 under flow.

In their active state, beta(2)-integrins, such as LFA-1, mediate the firm arrest of leukocytes by binding intercellular adhesion molecules (ICAMs) expressed on endothelium. Although the primary function of LFA-1 is assumed to be the ability to mediate firm adhesion, recent work has shown that LFA-1 can contribute to cell tethering and rolling under hydrodynamic flow, a role previously largely attributed to the selectins. The inserted (I) domain of LFA-1 has recently been crystallized in the wild-type (wt) and locked-open conformations and has been shown to, respectively, support rolling and firm adhesion under flow when expressed in alpha(L)beta(2) heterodimers or as isolated domains on cells. Here, we report results from cell-free adhesion assays where wt I-domain-coated polystyrene particles were allowed to interact with ICAM-1-coated surfaces in shear flow. We show that wt I-domain can independently mediate the capture of particles from flow and support their rolling on ICAM-1 surfaces in a manner similar to how carbohydrate-selectin interactions mediate rolling. Adhesion is specific and blocked by appropriate antibodies. We also show that the rolling velocity of I-domain-coated particles depends on the wall shear stress in flow chamber, I-domain site density on microsphere surfaces, and ICAM-1 site density on substrate surfaces. Furthermore, we show that rolling is less sensitive to wall shear stress and ICAM-1 substrate density at high density of I-domain on the microsphere surface. Computer simulations using adhesive dynamics can recreate bead rolling dynamics and show that the mechanochemical properties of ICAM-1-I-domain interactions are similar to those of carbohydrate-selectin interactions. Understanding the biophysics of adhesion mediated by the I-domain of LFA-1 can elucidate the complex roles this integrin plays in leukocyte adhesion in inflammation.

Binding Sites↗

Major determinants in hepatic disposition of polystyrene nanospheres: implication for rational design of particulate drug carriers.

The clearance of colloidal particles from the blood circulation occurs by phagocytes and/or endothelial cells, mainly in the liver, the spleen, and the bone marrow. The relative distribution of the injected particles in these organs is known to depend on various factors such as the size and surface properties of the particles and the type of serum proteins adsorbed onto the surface of particles. The basic principles behind their distribution characteristics into the reticuloendothelial system, however, remain unclear. This article reviews major determinants in hepatic disposition of polystyrene nanospheres, especially the relationship among physicochemical properties of the particle surface, the type of blood components associated onto the surface of particles, and their in vivo disposition characteristics in rats, and considerations to be given and implication for the rational design of particulate drug carriers are discussed.

Animals↗

Sulfoacylated macro-porous polystyrene-divinylbenzene low-capacity cation exchanger selective for amino acids.

A low-capacity cation-exchange column was newly developed for the separation of amino acids. A highly cross-linked macro-porous polystyrene-divinylbenzene co-polymer was functionalized by a sulfoacylation reaction. The exchange capacity was controllable at the acylation step. The capacity between 55 and 60 micromol/column was adequate for the practical separations in acceptable retention times. The 5-microm base polymers having average pore diameters smaller than 3 nm gave satisfactory results, and those having 1.5-nm pore was most favorable. Several isocratic elution conditions at different pH values adjusted by phosphate buffer of mM order with or without acetonitrile could provide good separations for individual classes of amino acids, i.e., acidic, neutral, hydrophobic, and basic groups. The results provided fundamental data for constructing gradient elution systems required for the simultaneous separation of protein amino acids.

Amino Acids↗

Low-capacity cation-exchange chromatography of amino acids using a novel sulfoacylated macroreticular polystyrene-divinylbenzene column with binary gradient elution.

This paper describes a versatile technique for amino-acid separation using a novel low-capacity sulfoacylated macroreticular polystyrene-divinylbenzene cation-exchange column with a simple binary high-pressure pH gradient elution. Proteinic 16 amino acids were well separated within 50 min using a H3PO4/Na2HPO4-CH3CN eluent system, and the cycle time was about 70 min. The chromatography with postcolumn OPA fluorescent detection was reproducible with RSDs less than 1% for retention times, and was quantitative with RSDs less than 5% for area responses. A linear regression line with an r2 value above 0.9990 was obtained for each analyte in concentration from 0.1 to 10 microM by 20 microL injection. The method was applicable to the separation and detection of urinary diagnostic amino acid due to inborn errors of metabolism, such as phenylketonuria. The analytical costs would be decreased by using the proposed method.

Amino Acids↗

Single-base mutation detection using neutravidin-modified polystyrene nanoparticle aggregation.

A single-base mutation assay using the non-crosslinking aggregation of neutravidin-modified polystyrene nanoparticles is described. This method requires only two steps: hybridization of biotinylated probe and sample DNAs, and then mixing with neutravidin-modified nanoparticles. The aggregation was detected within 20 min in total. A combination of the DNA non-crosslinking aggregation and biotin-avidin technology has allowed sufficient performance for the detection of single nucleotide polymorphisms.

Avidin↗

Rapid determination of decabromodiphenyl ether in polystyrene by thermal desorption-GC/MS.

A rapid determination of decabromodiphenyl ether (DeBDE) in polystyrene (PS) by thermal desorption (TD)-GC/MS was studied. The TD-GC/MS method using a pyrolysis-GC/MS system allowed the quick quantification of DeBDE in a waste TV back plate composing of a PS flame-retarded with polybrominated diphenyl ethers on the basis of the resulting chromatogram with a ca. 4% relative standard deviation without using any tedious sample pretreatment, such as solvent extraction.

Gas Chromatography-Mass Spectrometry↗

Synthesis of retro acyl carrier protein (74-65) fragment on a new glycerol based polystyrene support.

Retro-ACP (74-65) fragment was synthesized on a novel 4% tri-(propylene glycol) glycerolate diacrylate cross-linked polystyrene (PS-TRPGGDA) support. The peptide is grown from the functional site present in the cross-linker, which makes it unique and cost-effective among other styrene based polymer supports. A comparative study with Merrifield resin indicates high yield and purity of the peptide synthesized on the novel support.

Acrylates↗

A fundamental study of the microencapsulation procedure utilizing coacervation in a polystyrene-cyclohexane solution.

Coacervation in polystyrene (PS)-cyclohexane solution induced by the lowering of temperature was utilized to investigate the fundamental problems involved in the microencapsulation procedure. Polydispersity of PS played a vital role in determining variables at the critical state of phase separation, such as the composition of coacervate (dense) and lean phases. This also depended on temperature. Observations revealed that microcapsules of glass beads consist of a wall with a thin film of PS covered with a thick shell of talc. Poor utilization of PS may limit practical applications of this system unless effective measures are taken for the recovery of unutilized PS. Controlled release behaviour from microcapsules was successfully investigated by using encapsulated anhydrous sodium sulphate (ASS) particles, and applying the Higuchi model to the estimation of the effective diffusion coefficient of ASS through the composite wall. The values of diffusion coefficient decreased from an order of 10(-7) to 10(-8) cm/s by lowering the encapsulation temperature.

Cyclohexanes↗

Structure of hollow polystyrene microspheres: an SEM study.

Hollow polystyrene microspheres have been prepared by a modified interfacial polymer deposition technique. The hollow microspheres were characterized using scanning electron microscopy. The effects of polymer concentration and the quantity of the internal phase material (methanol) used on the microsphere size and their wall thickness were studied. Increase in polymer concentration causes increase in wall thickness while increasing internal phase material increases microsphere size and deforms them from a spherical shape.

Microscopy, Electron, Scanning↗

Inhibition of murine sarcoma cell adherence to polystyrene substrata by bacillus Calmette-Guérin: evidence for fibronectin-mediated direct antitumor activity of BCG.

Bacillus Calmette-Guérin (BCG) inhibited adherence of S180 mouse sarcoma cells and WI38 human diploid fibroblasts to the polystyrene substratum of 24-well cluster dishes in a dose-dependent manner. This property was retained by washed or heat-killed bacilli, but not by the vaccine filtrate or by the spent bacterial culture medium. Adhesion of bacilli to nonadherent S180 cells was demonstrated by light and scanning electron microscopy, but was not seen after trypsinization of adherent cells, indicating that bacilli bind to cell-surface adhesins. Preincubation of bacilli with human fibronectin abolished their ability to inhibit S180 adherence, suggesting that the phenomenon may be mediated by interaction of bacilli with cell-surface fibronectin. Fibronectin pretreatment of the bacteria also decreased their inhibition of S180 tumor growth in vivo, indicating that this mechanism may be at least partly responsible for BCG vaccine's observed antineoplastic activity.

Adsorption↗

Biliary excretion of polystyrene microspheres with covalently linked FITC fluorescence after oral and parenteral administration to male Wistar rats.

To further our understanding of microspheres as a drug delivery system, the biliary excretion of covalently linked FITC fluorescence polystyrene microspheres of various diameters was investigated after oral and parenteral administration to male Wistar rats. About 36%, 16% 3% and 1% of the dose was excreted into the bile after parenteral injection of 50 nm, 500 nm, 1 micron and 3 microns diameter microspheres, respectively, over 24 h. In addition, about 30%, 11%, 1%, and 1% of the dose, respectively, was found in the blood at 24 h. After oral administration of 50 nm, 500 nm, and 1 micron microspheres, the recovery was about 18%, 8% and 1% of the dose in the bile and about 9%, 1%, and 0% in the blood, respectively. No 3 microns microspheres were detected in the bile or blood after oral administration. Particles were not detected in the urine of any group tested. Thus, the present study demonstrated that biliary excretion plays an important role in the disposition of microspheres administered by the oral or parenteral routes, and there is a size-related excretion of microspheres into the bile.

Administration, Oral↗

Uptake by hepatocytes and biliary excretion of intravenously administered polystyrene microspheres in rats.

The in vivo uptake by hepatocytes and biliary excretion of fluorescein isothiocyanate-labeled polystyrene microsphere with a particle size of 50 nm (MS-50) after intravenous administration was studied in rats. It was confirmed by using confocal laser scanning microscopy that MS-50 was partially phagocytosed by the hepatocytes and that MS-50 taken up by the hepatocytes existed exclusively inside the cells 1 h after intravenous administration. Studies on the mechanism of the uptake of MS-50 by the hepatocytes using the liver perfusion technique revealed that a process mediated by apo-E was involved. After intravenous administration of MS-50, about 4% of dose was excreted into bile in 24 h. Pharmacokinetic evaluation of the excretion rate of MS-50 into bile showed that the process followed first-order kinetics. Qualitative evaluation of the fluorescence detected in the bile after intravenous administration of MS-50 revealed that the particles were certainly excreted into bile in an intact form. From these results, it was suggested that intravenously administered MS-50 would be partially phagocytosed by hepatocytes through a process mediated by apo-E and that MS-50 ingested by hepatocytes would be partially excreted into the bile.

Animals↗

Multi-Omics Platforms Reveal Synergistic Intestinal Toxicity in Tilapia from Acute Co-Exposure to Polystyrene Microplastics, Sulfamethoxazole, and BDE153.

Polystyrene microplastic (MP) and its co-existing contaminants may exert different toxic effects on its surrounding aquatic organisms. In order to detect the intestinal harmful responses, tilapia were subjected to exposure with 75 nm of MPs, 100 ng·L-1 of sulfamethoxazole (SMZ), 5 ng·L-1 of BDE153, and combinations thereof over periods of 2, 4, and 8 days. Enzymatic assays, transcriptomics, proteomics, and metabolomics were employed to evaluate intestinal histopathological effects. Results showed that significant reductions were observed in ATP, ROS, SOD, EROD, lipid metabolism-related enzymes, pro-inflammatory cytokines (TNFα and IL-1β), and apoptosis marker caspase 3 across all groups at day 8. Histological evaluation revealed diminished goblet cell density, with distinct vacuole formation in the BDE153+MPs group. KEGG pathway analysis highlighted disruptions in endocytosis, MAPK signaling, phagosome formation, and actin cytoskeleton regulation. Proteomic findings indicated notable enrichment in endocytosis (decreased sorting nexin-2; increased Si:dkey-13a21.4), MAPK/PPAR signaling, protein processing in the endoplasmic reticulum (Sec61 subunit gamma), and cytoskeletal modulation (reduced fibronectin; elevated activation peptide fragment 1), with or without SMZ and BDE153. Metabolomic profiling showed significant alterations in ABC transporters, aminoacyl-tRNA biosynthesis, protein digestion and absorption, and linoleic acid metabolism. In summary, these findings suggest that BDE153 and MPs synergistically exacerbate intestinal damage and gene/protein expression over time, while SMZ appears to exert an antagonistic, mitigating effect.

Animals↗