A direct colorimetric assay for Ca2+ -stimulated ATPase activity

A simple and rapid colorimetric assay for measuring the high affinity Ca2+-ATPase activity in subcellular fractions is presented. With this method a one-step addition of a malachite green/molybdate/polyvinyl alcohol reagent to the assay mixture at the end of the incubation period is all that is required for the spectrophotometric quantification of the phosphomolybdate-malachite green complex. The presence of polyvinyl alcohol allows the quantification of released phosphate without having to separate it from protein. We have validated this assay by characterizing the high affinity Ca2+-ATPase activity in isolated rat liver microsomes. Comparable Ca2+-ATPase activities in rat liver microsomes and adipocyte plasma membranes were found when measured with this colorimetric assay and an isotopic assay. This method is applicable to the measurement of other types of ATPase activities.     

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.     

Polyvinyl Alcohol with Cadmium Iodide and Fructose as an Aqueous Mounting Medium

An aqueous histologic and cytologic mounting medium containing polyvinyl alcohol, cadmium iodide and fructose is described. It is used in the preparation of permanent slides and may be applied to sections stained and rinsed in water or in any concentration of ethyl alcohol. The medium forms a hard, tough, quick-drying film which adheres well to the slide and cover glass. It is nonfluorescent, clear, optically homogeneous, and isotropic; it does not admit bubbles under the cover glass or crystallize on drying. It prevents or minimizes the bleeding of many stains from the sections; a list of 60 dyes, mostly basic, is given. The fresh medium has a pH of 4.4. The refractive index of the fresh medium is n_D²⁰ 1.4674; the dried film is n_D 1.6020 which may be lowered to n_D 1.5150 by decreasing the cadmium iodide in the formula. The viscosity at 25°C. is 7,299 centipoises. The medium has the following composition: distilled water, 40 g.; cadmium iodide, 34 g.; polyvinyl alcohol (Elvanol 51-05, du Pont's low viscosity grade) 18 g.; fructose, 8 g. The medium is prepared as follows: wash the polyvinyl alcohol with absolute methyl alcohol; dry and then grind in a mortar; dissolve the cadmium iodide in the water; add the polyvinyl alcohol while stirring with a speed-controlled motor-driven mixer; heat to 75°C. on a water bath with continuous stirring until dissolved; remove from the water bath and add the fructose while stirring; replace water lost by evaporation. The medium is ready for use when the foam has dispersed after standing.     

Water-soluble contrast media with increased viscosity for the diagnosis of diseases of the paranasal sinuses, bronchi and esophagus]

Water-soluble contrast agents with one of the polymers, cellulose derivatives or polyvinyl alcohol, were used in contrast examinations of paranasal sinuses, bronchi, and esophagus. Experiments have demonstrated the advantages of contrast agents basing on polymers as against the traditional oil-containing agents. Method for double-contrast examination of maxillary and frontal sinuses has been developed. Lowered concentrations of water-soluble contrast agents (i. e. diluted with distilled water) are recommended to be used, this permitting to save the reagents. A table of recommended dilutions of contrast media is presented. A total of 245 patients were examined, making use of contrast media of high viscosity.     

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.     

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.     

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.     

Synthetic polymers as xeno-free materials for stabilizing basic fibroblast growth factor in human mesenchymal stem cell cultures

Series of sulfonated polymers were evaluated as additives in cell culture media. Some of the compounds, such as sulfated polyvinyl alcohol (PVA), prevented denaturation and loss of basic fibroblast growth factor during cell culture and enhanced human mesenchymal stem cell proliferation. These compounds are xeno-free alternatives of heparin, an animal-derived sulfated saccharide, often used as an additive. To the best our knowledge, this study is the first to show that chemically defined synthetic chemicals, such as sulfated polyvinyl alcohol, can be used for this purpose.     

Catalytic Efficiency,Structure, and Recycling Behavior of Electrospun Polyvinyl Alcohol-Xylanase Fibers Cross-Linked by Glutaraldehyde

Food Biophysics - A versatile and effective method of producing polyvinyl alcohol (PVA)-xylanase (XY) fibers cross-linked by glutaraldehyde vapor (GA) is reported in the present study. Crosslinking...     

A Rapid Method for Making Permanent Mounts of Nematodes

A rapid method which can be used to mount and clear nematodes and their eggs is presented. Permanent mounts of certain nematodes and parasite eggs have been prepared using a medium consisting of 56 parts of a stock solution of polyvinyl alcohol (“PVA”), 22 parts phenol and 22 parts of lactic acid. The stock solution of PVA is prepared by dissolving 15 grams of PVA in 100 ml. of distilled water. This medium can be used on material killed and fixed in 10% formalin, any concentration of alcohol, alcohol-glycerin or glycerin. Results have been very satisfactory in most instances. An accompanying plate of photographs shows some of the preparations obtained by using this method.     

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。     

Some New Methods for Affixing Sections to Glass Slides. I. Aqueous Adhesives

As a new aqueous adhesive to affix sections to glass slides, hydrolyzed vinyl-triethoxysilane-either pure, in combination with polyvinyl alcohol or with specially prepared aqueous polyacrylate solutions-was applied. The silane proved to be very effective in enhancing bonding to the glass surface. As a general aqueous adhesive, a solution of 2% polyvinyl alcohol (m.w. 108,000; 99.7% hydrolyzed) with 0.2% hydrolyzed vinyltriethoxysilane is recommended. This stock solution is diluted 1:10 to 1:50 and used directly to float sections onto slides on a warming plate.     

Some new methods for affixing sections to glass slides. I. Aqueous adhesives

As a new aqueous adhesive to affix sections to glass slides, hydrolyzed vinyltriethoxysilane--either pure, in combination with polyvinyl alcohol or with specially prepared aqueous polyacrylate solutions--was applied. The silane proved to be very effective in enhancing bonding to the glass surface. As a general aqueous adhesive, a solution of 2% polyvinyl alcohol (m.w. 108,000; 99.7% hydrolyzed) with 0.2% hydrolyzed vinyltriethoxysilane is recommended. This stock solution is diluted 1:10 to 1:50 and used directly to float sections onto slides on a warming plate.     

Immobilization of invertase in polyvinyl alcohol coatings

A possibility of invertase immobilization in the polyvinyl alcohol coating formed directly on the electrode surface from water solution of polyvinyl alcohol and boric acid was being investigated. Conditions for obtaining the polymeric coating at the constant potential and at the constant current were compared. In order to obtain the polymeric coatings with a marked enzyme activity optimal conditions were found.     

Imaging and thermal studies of wheat gluten/poly(vinyl alcohol) and wheat gluten/thiolated poly(vinyl alcohol) blends

The morphology of wheat protein (WG) blends with polyvinyl alcohol (PVA) and respectively with thiolated polyvinyl alcohol (TPVA) was investigated by atomic force (AFM) and transmission electron microscopy (TEM) as well as by modulated dynamic scanning calorimetry (MDSC). Thiolated additives based on PVA and other substrates were previously presented as effective means of improving the strength and toughness of compression molded native WG bars via disulfide-sulfhydryl exchange reactions. Consistent with our earlier results, AFM and TEM imaging clearly indicate that the addition of just a few mole percent of thiol to PVA was sufficient to dramatically change its compatibility with wheat protein. Thus, TPVA is much more compatible with WG and phase separates into much smaller domains than in the case of PVA, although there are still two phases in the blend: one WG-rich phase and another TPVA-rich phase. The WG/TPVA blend has phase domains ranging in size from 0.01 to 0.1 microm, which are roughly 10 times smaller than those of the WG/PVA blend. MDSC further illustrates the compatibilization of the protein with TPVA via the dependence of the transition temperatures on composition.     

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.     

Preparation of DNA-polyintercalator conjugate and its functional property

Psoralen immobilized polyvinyl alcohol (PVA-P) was synthesized from chloromethylmethoxypsoralen and polyvinyl alcohol. The psoralen part of PVA-P intercalated into the double-stranded DNA and formed covalent bonding between the psoralen and nucleic acid base after 365 nm UV irradiation. As a result, DNA and PVA-P produced a water-insoluble conjugate. This DNA-PVA-P conjugate maintained the double-stranded structure of DNA and possessed the DNA's property, such as intercalation. Therefore, the DNA-PVA-P conjugate selectively accumulated the planar-structure containing chemical compounds, such as biphenyl and dibenzofuran, from an aqueous multi-component solution. These DNA-PVA-P conjugates may have the potential to be utilized as a separation material for the selective removal of harmful compounds.     

Brilliant Cresyl Blue as a Stain for Plant Chromosomes

Methods are proposed for staining plant chromosomes with the dye brilliant cresyl blue, and for making these stained preparations permanent by using polyvinyl alcohol mounting medium.

The stain, which is composed of 2% brilliant cresyl blue in 45% aqueous acetic or propionic acid, is used with fixed material in making smear preparations. The technics for staining are similar to those employed in the aceto-carmine method.

The mounting medium is made by mixing 56% polyvinyl alcohol, which is diluted in water to the consistency of thick molasses, with 22% lactic acid and 22% phenol by volume. The permanent slides are made by floating off the cover slip of the temporary slide in 70% alcohol, then applying the mounting medium and replacing the cover slip.

The chief advantages of the methods described are:

1)The preparation of the stain is rapid and simple. The batch of stain will be good with the first try.

2)The staining procedure in some instances is shorter than when using aceto-carmine.

3)The stain shows a high degree of specificity for nuclear structures and gives better results than aceto-carmine when used on certain plant tissues.

4)A minimum number of cells is lost in making the slides permanent when using polyvinyl alcohol mounting medium as the slide and cover slip are run through only one solution prior to mounting.

5)The mounting medium dries rapidly and this shortens the time required before critical examination of the permanent mounts can be made.     

Effect of deacetylation on wicking behavior of co-electrospun cellulose acetate/polyvinyl alcohol nanofibers blend

Cellulose acetate (CA) nanofibers webs deserve a special attention because of their very good water retention properties. CA nanofibers based biosensor in certain application come into contact with various liquids and requires high degree of wicking rate to transport liquid to its destination. Cellulose acetate (CA)/polyvinyl alcohol (PVA) blended nanofibers were prepared via co-electrospinning using double nozzle for jetting cellulose acetate and polyvinyl alcohol independently. The CA/PVA blend nanofibers webs were deacetylated in aqueous alkaline solution to convert CA in to regenerated cellulose and to remove PVA nanofibers from the raw web. The resultant nanofibers webs were characterized by wicking rate, water contact angle, SEM and FTIR analysis. The results revealed that by varying concentration of PVA solution enhances the wicking rate. Such a nanofibers web may be used in biosensor strip and other medical applications where the high wicking rates are desired.