Determination of low concentration of Paracoccus denitrificans encapsulated in polyvinyl alcohol LentiKat’s pellets

The aim of this work was to compare three methods to determinate low concentrations of Paracoccus denitrificans encapsulated in polyvinyl alcohol pellets, which is important for evaluation and optimization of pellet production as well as for monitoring of biomass growth. Pellets with different and well-defined biomass concentrations were used for experiments. The following fast and simple methods were tested: (1) dissolution of polyvinyl alcohol in hot water followed by dry weight estimation, (2) dissolution of polyvinyl alcohol in hot water followed by optical density measurement, (3) and extraction and quantification of proteins. Dry weight estimation proved to be problematic as it was difficult to separate biomass from polymeric carrier. Optical density measurement showed good linearity of dependence of optical density on biomass content, but determined limits of detection and limits of quantification were not within the range necessary for intended application. The only tested method meeting the requirements for sensitivity was determination of protein concentration after protein extraction.     

Stabilization of polyvinyl alcohol coating of polyester cloth for reduction of bacterial adhesion

Bacterial adhesion onto polyester cloth was greatly reduced by coating the cloth with polyvinyl alcohol (PVA). Under autoclaving conditions, the PVA coating was stable in water but not in 1% SDS. However, the treatment of the PVA-coated cloth with acidic 1% glutaraldehyde stabilized the coating to retain full resistance to bacterial adhesion even after autoclaving in 1% SDS.     

Preparation of films and fibers containing proteolytic enzymes

Immobilization of the proteolytic enzymes trypsin and terrylitin in fluorplastic-42 films and fibers as well as in water-soluble polymeric (polyvinyl alcohol and carboxymethyl cellulose) films was being studied during preparation of the films. The properties of the proteolytic enzymes immobilized in the polymers are dependent on the conditions of preparation of fibers and films.     

A quantitative study of the effects of different grades of polyvinyl alcohol on the activities of certain enzymes in unfixed tissue sections

Synopsis Different grades of the colloid stabilizer, polyvinyl alcohol, used for protecting unfixed cryostat sections during cytochemical reactions, may have different effects on enzymatic activity. The influence of three grades of polyvinyl alcohol on the activities of soluble, membrane-bound and membrane-enclosed enzymes has been investigated in unfixed sections; the activities were measured microdensitometrically.The largest molecular weight polyvinyl alcohol (G18/140, mol. wt. about 90 000) did not retain glucose-6-phosphate dehydrogenase activity in sections of rat liver even when used at the maximum convenient concentration (12%); G04/140 and M05/140 (molecular weights of 15 000 and 25 000 respectively) retained this soluble enzyme if used at concentrations of 30 and 20% respectively.At these concentrations, lactate dehydrogenase activity was apparently decreased when G04/140 and M05/140 were used; this diminished activity has been shown to be due to the need to establish optimal concentrations of reactants for each grade of polyvinyl alcohol and for each reaction. When optimal concentrations of reactants were used, the activities of this enzyme in the presence of each grade of polyvinyl alcohol were identical.The presence of any type of polyvinyl alcohol did not influence the activities of mitochondrial succinate dehydrogenase or of the smooth endoplasmic reticulum enzyme, 5,3-hydroxysteroid dehydrogenase. However, the presence of polyvinyl alcohol improved the state of the section.     

The effects of nonpenetrating cryoprotectants added to TEST-yolk-glycerol extender on the post-thaw motility of ram spermatozoa

The effects of adding five different concentrations of 17 polymeric compounds to TEST-yolk-glycerol extender on ram spermatozoa survival was studied. These were Aquacide (I, II, and III); dextran (0.8-1.6, 1.9, 15-20, 70, and 200-300 kDa); three types of Dri-Sweet; hydroxyethyl starch; methylcellulose, polyethylene glycol, polyvinyl alcohol, polyvinyl pyrrolidone, and Supercol 912. All the compounds tested except the Dri-Sweet compounds and hydroxyethyl starch significantly (P less than 0.05) decreased percentages of motile cells in unfrozen samples. The use of dextran (0.8-1.6 kDa; hydrolyzed dextran separated by ethanol) and Aquacide II significantly (P less than 0.05) increased post-thaw motility of spermatozoa frozen in pellets. Dextran (15-20 kDa), dextran (0.8-1.6 kDa), Aquacide II, and hydroxyethyl starch significantly (P less than 0.05) increased the percentages of post-thaw motility of ram spermatozoa frozen in the presence of glycerol and egg yolk.     

Preparation and evaluation of polyvinyl alcohol-co-oleylvinyl ether derivatives as tumor-specific cytotoxic systems

A series of poly(vinyl alcohol) amphiphilic derivatives have been prepared to obtain polymeric aggregates in aqueous phase holding thermodynamic instability. The aim was to evaluate their ability to interact with tumor cells eliciting selective cytotoxicity. The poly(vinyl alcohol) derivatives were prepared by partial substitution of poly(vinyl alcohol) (MW 10 kDa) with both oleyl chains and poly(ethylene glycol) monoethyl ethers (PEGMEE) of different molecular weights. The substitution degree was 1.5% for the oleyl chains and 1% for the PEGMEE chains (moles of substituent per 100 mol of hydroxyvinyl monomer). The polyvinyl derivatives obtained easily dissolved in water. Dynamic and static light scattering measurements on the polymer aqueous solutions indicated the formation of polymeric aggregates characterized by low polydispersity (0.232-0.299) and mean size (218-382 nm) in the range suitable for intravenous administration. Moreover, they were characterized by different packing densities and thermodynamic instabilities driving the polymers to interact with hydrophobic membranes. Among the analyzed polymers, the poly(vinyl alcohol)-co-oleylvinyl ether substituted with triethylene glycol monoethyl ether (P10(4)) provided in solution the highest affinity for hydrophobic membranes. P10(4), moreover, was the most cytotoxic toward the tumor cell lines analyzed (neuroblastoma: SH-SY5Y, IMR-32, HTLA-230. melanoma: MZ2-MEL, RPMI7932.), while it did not appreciably alter the viability of the normal resting lymphocytes. The peculiar behavior of the P10(4) aggregates has been correlated to their high thermodynamic instability in solution due to the high packing density that triggers the polymeric aggregates to interact with hydrophobic membranes such as the tumor cell membranes, thus eliciting cytotoxicity.     

Optimization of capillary coating by hydroxyethyl methacrylate for capillary zone electrophoresis of proteins

This work describes further improvements of coating fused silica capillaries with 2-hydroxyethyl methacrylate (HEMA) by atom transfer radical polymerization (ATRP). First, endcapping with a sterically less bulky silanyl reagent reduces the electrosmotic flow (EOF) by 25% in addition to the 40% EOF reduction caused by HEMA coating compared to a bare fused silica capillary. An additional hydrolysis step was introduced into the preparation of HEMA coated capillaries and leads to better reproducible migration times. The influence of the solvent during ATRP and the resulting polymer coating was investigated by replacement of DMF with water or water-methanol mixtures. The quality of the optimized coating was characterized by protein separations at pH 3. HEMA coated capillaries reveal up to 746000 plates. The polyvinyl alcohol (PVA) coated capillary provides only half of this efficiency. A long-term test at pH 9 shows good stability of the HEMA coated capillaries in basic medium. Also the numbers of plates in this medium was about 30% higher than for separations with the PVA capillary. In addition, the phosphate buffer was replaced by a volatile ammonium acetate buffer for later use with mass spectrometry (MS).     

The preparation of D-phenylalanine imprinted microbeads by a novel method of modified suspension polymerization

Molecularly imprinted polymeric microbeads (MIPMs) were prepared by the suspension and modified suspension polymerization methods using D-phenylalanine as the template, methacrylic acid as the functional monomer, ethylene glycol dimethacrylate as the cross-linker, toluene as the porogen, polyvinyl alcohol as the stabilizer, and sodium dodecyl sulfate as the surfactant. The addition of a surfactant to the conventional suspension polymerization mixture decreased the mean particle size of the MIPMs and increased the adsorption selectivity. For the modified suspension polymerization method, the mean particle size of the MIPMs was smaller than the particle size of MIPMs prepared via conventional suspension polymerization. Moreover, the adsorption selectivity improved considerably compared to the adsorption selectivities of MIPs reported previously.     

Comparison of pheromone application rates, point source densities, and dispensing methods for mating disruption of tufted apple bud moth (Lepidoptera: Tortricidae)

Small-plot (approximately 0.1 ha) studies were used to evaluate different pheromone dispensing systems, application rates, and point-source densities for mating disruption of the tufted apple bud moth, Platynota idaeusalis (Walker). Using polyvinyl chloride spirals impregnated with tufted apple bud moth pheromone (1:1 ratio of E11-tetradecenyl alcohol/E11-tetradecenyl acetate), pheromone rates of > or = 1,482 spirals per hectare (74.1 g pheromone per hectare) were superior to a rate of 988 spirals per hectare (49.4 g pheromone per hectare) in decreasing male response to pheromone traps in 1995, whereas no differences were detected among rates of 988, 1,482 and 1,975 spirals per hectare in 1996. Within a range of 370-988 pheromone dispensers per hectare, point source densities were equally effective in suppressing male response to pheromone traps. Pheromone-impregnated paraffin disks were equally effective at inhibiting male response to pheromone traps compared with polyvinyl chloride spirals. However, a paraffin emulsion formulation of pheromone applied with a hand-held grease gun provided longer residual communication disruption effects than polyvinyl chloride spirals. Dilution of paraffin emulsion pheromone formulations in water for application with a backpack sprayer and airblast sprayer rendered them ineffective in reducing male response to pheromone traps. The releases of pheromone from polyvinyl chloride spirals and paraffin disks aged in the field were described by a linear and negative logarithmic curve, respectively, indicating that dispenser life time should be longer for spirals. The ratio of acetate to alcohol components of pheromone released from spirals increased over time, whereas the release ratio remained more constant for paraffin disks. This suggests that the disruption efficacy of spirals may be prematurely reduced because of imbalance of the released components.     

Polyvinyl alcohol--trypsin as a catalyst for amino acid ester synthesis in organic media

The bovine trypsin-catalyzed synthesis of N-alpha-benzoyl-DL-arginine esters from N-benzoyl-DL-arginine were studied in various organic solvents. Trypsin was immobilized to polyvinyl alcohol (PVA) by adsorption from its aqueous solutions. Immobilized enzyme showed higher catalytic activities than free enzyme for amino acid esterification in ethanol. The yield of ester is strongly dependent upon the PVA/trypsin ratio and water content in the reaction medium. The rate and equilibrium constant of the ester formation reaction are also dependent on water content.     

Decoration of electrical conductive polyurethane-polyaniline/polyvinyl alcohol matrixes with mussel-inspired polydopamine for bone tissue engineering

Electrospinning is a versatile technology for the fabrication of nanofibrous matrixes to regenerate defects. This study aims to develop a functionalized and electroconductive polymeric matrix to improve rat bone marrow mesenchymal stem cell adhesion, proliferation, and differentiation. Herein, the influence of the chemical composition of the substrate on homogeneous modification of the surface with mussel-inspired polydopamine (PDA) is focused. Accordingly, the deposition of PDA on the surface was proved by Fourier transform infrared spectroscopy. Morphologies of the scaffolds demonstrated homogeneous decoration of the polyvinyl alcohol (PVA)/polyurethane (PU)-polyaniline (PANI) matrixes with PDA, while a lower density of mussel-inspired polymer was observed in bare PU-PANI constructs. Although uniform and dense precipitation of PDA reduced conductivity of scaffolds 1.2 times compared with the samples with a low density of the PDA, 1.1 and 1.2 times enhancement in tensile strength and Young's modulus, respectively, were the strength of the applied process, especially in bone tissue engineering area. Contact angle measurements demonstrated about two times reduction in measured values, which shows improvement in hydrophilicity of PDA-modified PVA/PU-PANI fibers compared with PDA-coated PU-PANI ones. Swelling ratio and mass loss ratio calculations revealed enhancement in measured values as a function of homogeneous and dense coating, which arise from hydrophilicity of the polymeric substrate. The bioactivity test indicated that a dense layer of PDA strongly supports formations of hydroxyapatite-like crystals. Moreover, homogeneous decoration of conductive matrixes with PDA showed suitable cell viability, adhesion, and spreading while cell-scaffolds interactions improved under electrical stimulation. Higher expression of alkaline phosphatase and secretion of Collagen I under the electrical field proved the applicability of modified electroconductive scaffolds for further preclinical and clinical studies to introduce as a reconstructive bone substitute.     

Immobilization of enzymes on spongy polyvinyl alcohol cryogels: the example of beta-galactosidase from Aspergillus oryzae.

The catalytic properties of a beta-galactosidase from Aspergillus oryzae, entrapped into a spongy polyvinyl alcohol cryogel, were studied. This polymeric matrix was selected because of its mild conditions of preparation and its stability, biocompatibility, structural strength and diffusive properties. The enzyme was entrapped, in high percentage, into cryogel sponges and its activity and kinetic parameters were determined and compared with those of the free enzyme, using as substrates o-nitrophenyl-beta-galactopyranoside (ONPG) or lactose. The immobilized enzyme showed a reduced activity with ONPG and lactose, probably because of substrate diffusion limitations through the matrix, but it was more stable to temperature, pH and ionic strength than the free enzyme. Lactose hydrolysis under continuous experimental conditions was performed using the matrix-enzyme cited above.     

Storage of industrial microorganisms entrapped into polymer matrices

Our study of the techniques of long-term storage of the biomass of various strains of microorganisms, which cause breakdown or transformation of synthetic organic compounds, demonstrates that desiccated agar beads with immobilized microbial cells can be used for this purpose. In addition, the cells can be stored in desiccated matrices of agar or polyvinyl alcohol, coating synthetic cords. Such dry biocatalysts may be used for quick starting of bioreactors and in other biotechnological processes. The technique is applicable to storage of various strains of Pseudomonas, Corynebacterium, Rhodococcus, and, to a lesser extent, Enterobacteriaceae.     

白蚁纤维素粉饵料成型技术研究

为研究白蚁饵料成型工艺,比较了9种胶黏剂对白蚁纤维素饵料成型效果、耐水性能以及对白蚁取食的影响。结果表明:20%、40%剂量糊精;50%、100%剂量三聚氰胺甲醛树脂,10%、20%、40%田菁胶、卡拉胶、壳聚糖、明胶;10%、50%、100%聚乙烯醇、硅酸钠对微晶纤维素的成型效果较好,经上述剂量胶黏剂处理后,纤维素饵块的邵氏硬度(HD)极显著高于对照。耐水性能试验至第30天时,50%聚乙烯醇、100%聚乙烯醇、100%三聚氰胺甲醛树脂处理的纤维素饵块的溃散程度指数分别为1.33、1.00、2.00,其余饵块的溃散程度指数均达3级。生测结果显示,在7 d的室内强迫取食试验中,白蚁对50%聚乙烯醇、100%聚乙烯醇、100%三聚氰胺甲醛树脂处理的块状纤维素饵料的取食率均极显著低于对照,说明饵块中添加的上述胶黏剂对白蚁的取食具有一定的影响。综上,50%聚乙烯醇、100%聚乙烯醇、100%三聚氰胺甲醛树脂适用于白蚁纤维素饵料成型,但若想获得白蚁喜食的饵块仍需对配方做进一步的优化。     

Polymer shelled microparticles for a targeted doxorubicin delivery in cancer therapy

Targeting is a main feature supporting any controlled drug delivery modality. Recently we developed poly(vinyl alcohol), PVA, based microbubbles as a potential new ultrasound contrast agent featuring an efficient ultrasound backscattering and a good shelf stability. The chemical versatility of the polymeric surface of this device offers a vast variety of coupling modalities useful for coating and specific targeting. We have designed a conjugation strategy on PVA shelled microbubbles to enable the localization and the drug delivery on tumor cells by modifying the surface of this polymeric ultrasound contrast agent (UCA) with oxidized hyaluronic acid (HAox). After the conversion of the microbubbles into microcapsules, the kinetics of the release of doxorubicin, a well-known antitumor drug, from uncoated and HAox-coated PVA microbubbles and microcapsules was investigated. Cytocompatibility and bioadhesive properties of the HA-modified microparticles were then tested on the HT-29 tumor cell line. Cytotoxicity to HT-29 tumor cells of microcapsules after loading with doxorubicin was studied, evidencing the efficacy of the HAox coating for the delivery of the drug to cells. These features are a prerequisite for a theranostic, that is, diagnostic and therapeutic, use of polymer-based UCAs.     

Tissue stabilisation during histochemical reactions: The use of collagen polypeptides

Summary The incorporation of polyvinyl alcohol into the incubation medium prevents the loss of material from unfixed tissue sections during incubation. Polypeptides, derived from the partial degradation of collagen, are equally effective in retaining this material; at the same time they offer certain advantages over polyvinyl alcohol as tissue stabilisers.I wish to thank Mr. G. Frost for providing the polypeptides and Dr. J. Chayen for his interest and advice. I am grateful to the Medical Research Council for a grant and the Arthritis and Rheumatism Council for Research for general support.     

The clinical, histologic, and ultrastructural presentation of polyvinyl sponge (Ivalon) breast prostheses removed for massive fluid accumulation.

The current study describes what we believe is the first report of bilateral massive seromas associated with open-cell Ivalon sponges. Additionally, the gross, histologic, and ultrastructural features consistent with previous reports of polyvinyl alcohol prostheses are presented. Despite the reported chemical inertness of polyvinyl alcohol, this material may incite a biologic response in some patients, leading to dense fibrosis and occasional foreign-body giant-cell reaction. It is postulated that the molecular breakdown products of the polyvinyl alcohol polymer may create an osmotic gradient across the periprosthetic capsule, which may lead to intracapsular fluid accumulation, as presented in this case.     

Storage of the strains of industrial microorganisms, entrapped in polymer matrices

Our study of the techniques of long-term storage of the biomass of various strains of microorganisms, which cause breakdown or transformation of synthetic organic compounds, demonstrates that desiccated agar beads with immobilized microbial cells can be used for this purpose. In addition, the cells can be stored in desiccated matrices of agar or polyvinyl alcohol, coating synthetic cords. Such dry biocatalysts may be used for quick starting of bioreactors and in other biotechnological processes. The technique is applicable to storage of various strains ofPseudomonas, Corynebacterium, Rhodococcus, and, to a lesser extent, Enterobacteriaceae.     

Thermal inactivation and stabilization of lysozyme substrate-- Micrococcus lysodeicticus cells

Heat inactivation of the acetonic powder of Micrococcus lysodeicticus cells suspended in phosphate buffer pH 6.2 was quantitatively characterized in the temperature range from 34 to 52 degrees. The total value of the rate constant for heat inactivation of the cells equals 2.88 X 10(8) exp(-18360/RT) sec-1. The activation parameters of the process at 34 degrees are the following: delta H* = 17.7 kcal/mole; delta S* = 21.8 E. U.; delta F* = 24.4 kcal/mole. The effect of ethylene glycol, mannitol, dextran, polyvinyl alcohol (PVA) and polyethylene glycols with different molecular weights on the lysis rate and cell stability was studied. Polyvinyl alcohol was found to be the most effective stabilizer. At concentrations of about 10(-5) it enhances the thermostability of the cells threefold.     

Luminescent light diffuser for diffuse lighting applications

The lighting industry currently accounts for a significant proportion of all energy demand. Luminescent white lighting is often impure, inefficient, expensive, and detrimentally emits as a point source, meaning the light is emitted from a focused point. A luminescent light diffuser offers the potential to create a spatially broad lighting fixture. We developed a luminescent light diffuser consisting of three commercially available luminescent dye species (rhodamine 6G, fluorescein, 7-diethylamino-4-methylcoumarin) dispersed within a polymer matrix (polyvinyl alcohol), or commercial paint, and coated on a planar waveguide. A Light-emitting diode (LED) (385 nm) is directed into the waveguide which excites the luminescent species, coating the panel, creating a device that emits spatially broad pure white light. As the emission depends on escape cone emission from the waveguide, the device’s emission was found to depend highly on the coating film quality and components. We present two systems: a small 40 mm × 40 mm prototype, made using standard water-soluble polymer (polyvinyl alcohol), to study the underlying operational principles, and a 100 mm $\times$ 100 mm device with optimized efficiency fabricated with a clear commercial paint. By doping the polymer matrix with scattering silica microparticles we achieved a maximum photon outcoupling efficiency of 78%, whilst maintaining colour purity with an increased device size of more than 300 times (compared with the input LED). This work shows that it is possible to construct an inexpensive and spatially broad lighting source, whilst maintaining colour purity at a low cost.