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[Serologic-nephelometric method for the detection of allergic reactions to small-molecular drugs using polystyrene sodium particles instead of soluble polystyrene sulfonate sodium as pharmaceutic aids].

The previously developed two-step method using soluble polystyrene sulfonate-Na for the detection of allergic reactions to small molecular drugs has been replaced by a one-step method using particles of the same substance. The new method is simpler to perform. Particulate polystyrene sulfonate is commercially available. The drug is added in increasing concentrations to diluted serum samples and the light scattering is measured with a highly sensitive nephelometer. Both the specific serum factor in the patient's serum (globulins) and the non-specific factor contained in most fresh human sera are particularly labile.

Antibody Formation

New useful reagents for peptide synthesis. Insoluble active esters of polystyrene-bound 1-hydroxybenzotriazole.

Insoluble 1-hydroxbenzotriazole-bound polystyrene was prepared through a series of chemical modifications of commercially available polystyrene. Reaction of 3-nitro-4-chlorobenzyl alcohol or of 3-nitro-4-chlorobenzyl bromide with polystyrene in the presence of aluminium trichloride yielded (3-nitro-4-chloro)benzylated polystyrene. Upon reaction with hydrazine it was converted to (3-nitro-4-hydrazine) benzylated polystyrene which was cyclized, by acidolysis, to yield 1-hydroxybenzotriazole-bound polystyrene. This was coupled, using N, N' -dicyclohexylcarbodiimide as the coupling agent, to many N-blocked amino acid derivatives, yielding polymeric polystyrene-bound active esters. Such derivatives are highly reactive and their efficacy in the synthesis of several peptides, including that of the tetrapeptide Boc-L-Leu-L-leu-L-Val-0bzl-L-Tyr-0Bzl and of thyrotropin-releasing hormone was demonstrated.

Dipeptides

Interrelationships between soil microorganisms and polystyrene.

Transmission and scanning electron microscopy along with autoradiographic procedures were used for evaluating the interrelationship between soil microorganisms and polystyrene. Two hypotheses, allowing to explain the properties of polystyrene as a soil conditioner, were investigated: the first is concerned with a possible action of soil microorganisms on the compound; the second, reciprocally, with the polystyrene interference on microorganisms. Radioactivity translocation of 14C-Ecolyte-polystyrene along fungal hyphae and asexual fructification of strains, isolated from soil, as well as cytological modification at the cell wall level of the same microfungi, cultivated in the presence of polystyrene have been ascertained.

Cladosporium

On the constancy in composition of polystyrene and polymethylmethacrylate plastics.

Variations in the atomic composition, and mass and electron densities of polystyrene and polymethylmethacrylate (PMM) plastics were assessed from experimentally determined mass attenuation coefficients for 125I and 137Cs gamma rays. The means and standard deviations in the mass densities of 16 samples of PMM and 10 samples of polystyrene were found to be 1.174 +/- 1.4% and 1.042 +/- 0.6% g/cm3, respectively. Based upon transmission measurements on various solutions of ethyl alcohol in water, the standard deviations in the effective atomic numbers of PMM and polystyrene were determined to be 0.77% and 1.3%, respectively. Based upon experimentally determined mass attenuation coefficients for 137Cs, the standard deviations in electron density for PMM and polystyrene were 0.5% and 1.2% respectively. Similar measurements on tap water and two grades of distilled water failed to detect any differences in atomic composition.

Methylmethacrylates

Superoxide production from human polymorphonuclear leucocytes stimulated with immunoglobulins of different classes and fragments of IgG bound to polystyrene dishes.

Polystyrene surfaces coated with proteins at alkaline pH were found to be useful to investigate the stimulation of neutrophils independently of phagocytosis. Human neutrophils, when exposed to appropriate stimuli, release superoxide anion (O2-). We have measured superoxide dismutase-inhibitable cytochrome c reduction by cells stimulated with various kinds of protein coated on polystyrene dishes. Native immunoglobulins in solution did not stimulate O2- generation in neutrophils. IgG and IgA adherent to polystyrene dishes stimulated O2- generation in neutrophils, but IgM, IgD, IgE, albumin, Fab, and Fc of IgG on polystyrene dishes did not.

Humans

Separation of transfer ribonucleic acids on polystyrene anion exchangers.

The transfer RNA separation by chromatography on strong-base polystyrene exchange materials is examined and compared with the widely used reversed-phase chromatography. Results indicate important differences in some transfer RNA (tRNA) elution patterns by the anion-exchange chromatography, as compared with the reversed-phase chromatography. Transfer RNAs containing hydrophobic groups are adsorbed more strongly. The anion exchanger has twice the number of theoretical plates. Single peaks of tRNA2Glu and tRNA1Phe obtained from the reversed-phase column give multiple peaks only polystyrene anion-exchange chromatography. All six leucine tRNAs (Escherichia coli) and differences in tRNA populations synthesized during early and late stages of the dividing lymphocytes from normal human blood can be characterized by the anion-exchange chromatography. Different separation profiles are obtained by two separation systems for tyrosine tRNAs from mouse liver and mouse-plasma-cell tumor. The results indicate that, in contrast to the reversed-phase chromatography, strong-base-polystyrene anion-exchange chromatography is capable of separating tRNAs with minor structlral differences.

Animals

Polystyrene embedding: a new method for light and electron microscopy.

Polystyrene embedments of histological specimens can be obtained with a solution of 1:14 polystyrene-toluene, 5% benzyl alcohol and 1% dibutyl phthalate, allowing the solvent to evaporate in polyethylene containers for 2-3 days at 58 C. The resulting blocks are easily cut into truncated pyramids, each containing a piece of tissue, which are then glued to a Plexiglas support. Drying is completed at 80 C for 20 hr. The pyramids can then be sectioned to produce thick sections with a steel knife or to produce semi- or ultrathin sections with a glass knife. A 10% paraldehyde solution is used to mount the light microscopy sections on a slide heated on a hot plate to 80 C; these can be treated with the same techniques used with paraffin sections. The results are of high quality. Semithin sections of tissues fixed for electron microscopy can be stained directly after mounting, or by a wider range of stains once the polystyrene has been removed by organic solvents. In electron microscopy, the ultrathin sections obtained with the usual techniques are highly electron beam-resistant and given acceptable results.

Animals

Immobilization of viral and mycoplasma antigens and of immunoglobulins on polystyrene surface for immunoassays.

The immobilization of purified influenza virus, rubella virus, crude nuclear cytomegalovirus antigen and of mycoplasma on polystyrene tubes was studied using radio-iodinated preparations. The antigen activities on tube surfaces were determined using sequentially specific human antibodies and alkaline phosphatase-conjugated anti-human IgG in an enzyme-immunoassay (EIA) reaction. In addition, immobilization of radio-iodinated human IgG, IgM, and IgA, serving as model proteins, was studied using the respective anti-immunoglobulin conjugates in EIA directly. Pretreatment of the surface with albumin and glutaraldehyde inhibited the adsorption and antigenicity of IgG. Increase of temperature and thus of speed of adsorption did not affect the fraction of antigen eluted during the test procedure. Only with IgG and IgA was it necessary to saturate the polystyrene surface in order to achieve maximal reactivity in EIA. With other antigens, maximal reactivity in EIA was obtained with amounts of protein much lower than the maximal amount that could be adsorbed per tube. IgM was found to have an exceptionally high affinity to polystyrene.

Antigens, Bacterial

Sulphonated polystyrene as an optimal substratum for the adhesion and spreading of mesenchymal cells in monovalent and divalent saline solutions.

Cell adhesion and spreading were studied on sulphonated polystyrene dishes in serum-free saline (Mn, Na, Cl, buffer) i.e., without an intervening protein layer. Spreading as a function of surface charge density, SCD, peaked around 2-10 negative charges per square nanometer, corresponding to a monomolecular layer of sulphonate ions. At optimal SCD, macrophages, BHK-C13 and whole mouse embryo secondary cells all showed considerable spreading, even in monovalent saline-more so than on a conventional tissue-culture surface. But outside this narrow range of SCD, or on protein-coated surfaces, the divalent cation was indispensable. The biphasic effect of sulphonation on cell adhesion is consistent with the theory that a substratum need not be biochemically specific, provided it is physiochemically polar, rigid and dense. According to this theory, polystyrene of sub-optimal SCD would not be sufficiently polar, while supra-optimal sulphonation would produce a hydrogel surface, lacking in local rigidity and density, due to osmotic swelling. The principle of polymer exclusion, by a surface hydrogel layer, is also consistent with observations on the inhibitory effects of adsorbed proteins-viz., albumin, collagen, serum and cellular exudate, respectively-contrasted with the ready attachment of cells to a bare, optimally charged substratum, in this minimal in vitro system.

Animals

Polystyrene balls as the solid-phase of a double-antibody radioimmunoassay for human serum albumin.

Polystyrene balls have been incorporated as the solid-phase of a model double-antibody radioimmunoassay for human serum albumin. Purified IgG from the secondary antiserum is adsorbed on the 6.4 mm diameter balls. The solid-phase secondary antibody is then used to separate primary antibody bound iodinated antigen from unbound antigen. The secondary antibody coated polystyrene balls are easily prepared and manipulated; several hundred sample dilutions can readily be processed in a single assay. Assay background values of 1.5% or less are consistently obtained without extensive or special washing procedures.

Animals

Scanning electron microscopic appearance of viral-antigen-coated polystyrene balls.

A radioimmunoassay (RIA) was recently developed for the detection of antiviral IgG and IgM class-specific antibodies using antigen-coated polystyrene balls as the RIA solid-phase. In this communication the attachment and distribution of herpes simplex virus (HSV) capsid and envelope antigens and rubella viruses on the surface of the balls was examined by scanning electron microscopy (SEM). In SEM the surface of the untreated 'clear frosted' polystyrene balls appeared very uneven with innumerable pits and grooves. The viral particles were haphazardly distributed both in the grooves and on the exposed surface of the balls. The strength of adsorption of the viral antigens onto the balls seemed to be remarkably resistant to outside mechanical forces. HSV antigens frequently appeared in clusters, whereas rubella viruses were mostly found as single particles.

Antigens, Viral

UVB-aged polystyrene microplastics induce enhanced stress responses in human proximal tubular cells.

Microplastics (MPs) are increasingly detected in human biological matrices, raising concerns about their potential systemic effects, including on the kidney. However, the cellular responses of renal tubular epithelium to MPs and the role of environmental aging processes in modulating their biological activity remain poorly defined. Under environmental conditions, MPs undergo photo-oxidative transformations that alter their surface chemistry and may influence their interactions with biological systems. In this study, we investigated the effects of 1 µm polystyrene MPs in virgin (MPsV) and UVB-oxidised (MPsOx) forms in a human renal proximal tubular cell line (HK-2). Cells were exposed to MPs (25 and 50 µg/mL), and multiple endpoints related to cellular stress and genomic stability were evaluated, including lysosome-associated responses, oxidative damage, DNA integrity, micronucleus formation, DNA-content distribution profiles as an indirect proxy of proliferative status, and cytoskeletal organisation. Exposure to MPs induced measurable stress responses in tubular cells, with oxidised particles generally eliciting stronger effects than MPsV. These responses were consistent with increased oxidative stress, lysosome-associated cellular responses, genomic instability-associated alterations, activation of stress-responsive molecular pathways, and cytoskeletal perturbation. Collectively, these findings indicate that environmentally aged polystyrene MPs elicit more evident cellular stress responses than their virgin counterparts in HK-2 cells. Our results highlight the importance of incorporating environmentally transformed MPs into toxicological testing frameworks to improve the biological relevance of hazard assessment.

Genotoxicity

Microbial degradation of [C14C]polystyrene and 1,3-diphenylbutane.

Microbial degradation of [beta-14C]polystyrene and 1,3-diphenylbutane, a compound structurally representing the smallest repeating unit of styrene (dimer), was investigated in soil and liquid enrichment cultures. Degradation rates in soil, as determined by 14CO2 evolution from applied [14C]polystyrene, varied from 1.5 to 3.0% for a 4-month period. Although relatively low, these percentages were 15 to 30 times greater than values previously reported. Enrichment cultures, containing 1,3-diphenylbutane as the only carbon souce, were used to determine the mechanisms of microbial oxidation of the polymer chain ends. Metabolism of 1,3-diphenylbutane appeared to involve the attack by a monooxygenease to form 2-phenyl-4-hydroxyphenylbutane followed by a further oxidation and subsequent fission of the benzene ring to yield 4-phenylvaleric acid and an unidentified 5-carbon fragment via the classic meta-fission pathway. Phenylacetic acid was probably formed from 4-phenylvaleric acid by subsequent beta-oxidation of the side chain, methyl-oxidation and decarboxylation. An initial examination of the population of microorganisms in the diphenylbutane enrichment cultures indicated that these oxidative reactions are carried out by common soil microorganism of the genera Bacillus, Pseudomonas, Micrococcus, and Nocardia.

Bacteria

Quantitative analysis of styrene monomer in polystyrene and foods including some preliminary studies of the uptake and pharmacodynamics of the monomer in rats.

A variety of food containers, drinking cups and cutlery, fabricated from polystyrene (PS) or polystyrene-related plastic, were analyzed for their styrene monomer content. Samples of yogurt, packaged in PS cups, were similarly analyzed and the leaching of styrene monomer from PS containers by some food simulants was also determined. Blood level studies with rats, dosed with styrene monomer by various routes, illustrated uptake phenomena that were dependent on the dose and route of administration and were also affected by the vehicle used to convey the styrene monomer.

Animals

Platelet retention by albuminated glass and polystyrene beads.

Ex vivo platelet retention by albuminated glass and polystyrene beads has been evaluated as a function of flow rate, bead surface area, blood exposure time and albumin treatment. The stability of the albumin coatings as well as scanning electron microscopy of the various surfaces before and after blood exposure has also been included. Results indicate that platelet retention is sensitive to changes in the above parameters and that albumin pretreatment of different substrates can decrease platelet retention. This decrease is substrate dependent in that platelet retention is different for the albuminated glass and polystyrene substrates. Chemical analysis of the substrate materials by X-ray photoelectron spectroscopy (XPS) as well as bulk chemical analysis is also reported.

Animals

Mutagenicity study of workers employed in the styrene and polystyrene processing and manufacturing industry.

Mutagenicity studies were undertaken on lymphocytes from the following groups of persons exposed to styrene or polystyrene: one group from a styrene manufacturing plant, one from a polystyrene plant, and three from plants processing unsaturated polyester resins with different manufacturing methods. For all the groups the concentration of styrene in the atmosphere of the workplace was measured at various points and the amount of urinary mandelic acid was determined. In each case 100 metaphases were analyzed for chromosomal aberrations. The results were compared with reference groups.

Adult