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Preparation of peanut butter suspension for determination of peanuts using enzyme-linked immunoassay kits.

Peanuts are one of the 8 most common allergenic foods and a large proportion of peanut-allergic individuals have severe reactions, some to minimal exposure. Specific protein constituents in the peanuts are the cause of the allergic reactions in sensitized individuals who ingest the peanuts. To avoid accidental ingestion of peanut-contaminated food, methods of analysis for the determination of the allergenic proteins in foods are important tools. Such methods could help identify foods inadvertently contaminated with peanuts, thereby reducing the incidence of allergic reactions to peanuts. Commercial immunoassay kits are available but need study for method performance, which requires reference materials for within- and between-laboratory validations. In this study, National Institute of Standards and Technology Standard Reference Material 2387 peanut butter was used. A polytron homogenizer was used to prepare a homogenous aqueous Peanut Butter suspension for the evaluation of method performance of some commercially available immunoassay kits such as Veratox for Peanut Allergen Test (Neogen Corp.), Ridascreen Peanut (R-Biopharm GmbH), and Bio-Kit Peanut Protein Assay Kit (Tepnel). Each gram of the aqueous peanut butter suspension contained 20 mg carboxymethylcellulose sodium salt, 643 microg peanut, 0.5 mg thimerosal, and 2.5 mg bovine serum albumin. The suspension was homogenous, stable, reproducible, and applicable for adding to ice cream, cookies, breakfast cereals, and chocolate for recovery studies at spike levels ranging from 12 to 90 microg/g.

Arachis↗

Hypersensitivity reactions to vaccine components.

Vaccines are responsible for the control of many infectious diseases that were once common in the United States, including polio, measles, diphtheria, pertussis (whooping cough), rubella (German measles), mumps, tetanus, and Haemophilus influenzae type b. National efforts to generate collaboration between federal, state, and local governments and public and private health care providers have resulted in record high levels of vaccination coverage in the United States. The high rate of US vaccinations is paralleled by growing concerns about the safety of their delivery. The variety of substances used in vaccines sometimes causes the development of cutaneous reactions in susceptible adults and children. This article will review adverse cutaneous events consistent with hypersensitivity reactions to the following ingredients in vaccines: aluminum, thimerosal, 2-phenoxyethanol, formaldehyde, and neomycin.

Aluminum↗

Allergic contact dermatitis to preservatives.

In summary, a wide variety of skin care products contain preservatives. Patients who are allergic to one of these preservatives may have either localized or widespread dermatitis. Affected patients may find it difficult to avoid thimerosal without the help of the health care provider because the use of these allergens is so widespread. Patch testing is an invaluable tool for patients who struggle with dermatitis. Antigen-avoidance lists that facilitate patient education about what products to avoid are available from the manufacturers of patch test allergens (for example, TRUE Test or Chemotechnique). These lists are helpful starting points for patients in that they provide general categories (for example, shampoos, soaps, or creams) of products that the patient should avoid. With these printed guidelines alone, patients must read skin care product labels carefully, looking for the names of their allergens as identified by patch tests as well as for any synonyms and cross-reactors of these allergens. Thus, patients may feel overwhelmed by hearing the names of allergens that are long and complex. After an allergen has been identified, the nurse can play a key role in helping patients understand their dermatitis and its management. Nurses are in a unique position to spend time educating patients about how to uncover the sources of specific allergens and, subsequently, how to avoid them. The Contact Allergen Replacement Database can help in this educational process by giving patients a shopping list of specific items that are free of the specific allergens causing their allergic contact dermatitis.

Causality↗

[The chromosome-damaging action of 2 chemical vaccine contaminants on mice].

The cytogenetic effects of two chemical agents, hydroxylamine used for the destruction of bacterial cells and thimerosal added to many immunobiological preparations as preservative, were studied in vivo by their action on the marrow cells of C57BL/6J mice. The preparations under study, when injected intraperitoneally in a wide range of doses, including subtoxic ones, induced no chromosomal aberrations. At the same time cyclophosphamide, an antitumor cytostatic agent used for positive control, produced a pronounced damaging effect on chromosomes.

Animals↗

Survival of Acanthamoeba in contact lens rinse solutions.

Acanthamoeba may cause a severe keratitis in contact lens wearers. Since most sterilization techniques require rinsing the lenses prior to insertion, contaminated solutions represent a potential vector for transmission of Acanthamoeba. The ability of rinse solutions to sustain an inoculum of Acanthamoeba polyphaga was investigated. A. polyphaga was exposed to 0.1% benzalkonium chloride, 0.001% thimerosal/0.1% edetate disodium, 0.1% edetate disodium, saline, tap water, and distilled water. The status of the organism was evaluated with direct microscopic counts and cultures to confirm viability. Incubation with 0.1% edetate disodium, saline, tap water, and distilled water resulted in the maintenance of reduced populations of viable organisms for 7 days. Benzalkonium chloride preserved saline and solutions containing thimerosal with edetate rendered the Acanthamoeba nonviable.

Acanthamoeba↗

[Lethal mercury poisoning due to ingestion of merthiolate].

A case of mercurial poisoning caused by ingestion of an organic mercurial compound, thimerosal, found in local antiseptic solutions. The clinical picture consisted of grave neurological symptoms which were not reversed by penicillamine and resin administration despite rapid plasma level reduction of mercury. We call attention to this case because of the widespread availability of antiseptic solutions containing mercurial compounds and alcohol.

Adult↗

Preservative effects on the healing rate of rabbit corneal epithelium.

Benzalkonium chloride (BAK), sodium lauryl sulfate (SLS), chlorhexidine digluconate, thimerosal and polyaminopropyl biguanide were tested for their effects on the rate of corneal epithelial regrowth after mechanical lesions, and BAK and SLS after heptanol-generated lesions in rabbit corneas. Only 1.3% SLS reduced the rate of epithelial healing after three times a day drop application as determined using planar measures of epithelial regrowth. SLS, a common ingredient in soups and shampoos, should be used carefully where a lesion or wound is present in the corneal epithelium.

Animals↗

[Cytogenetic control over the safety of merthiolate].

The capacity of merthiolate (also known as thimerosal), a mercury organic compound used as preservative in vaccines, to induce chromosomal aberrations was studied in vivo in experiments on mice. The metaphasic method of the registration of anomalies was used. In a dose of 3.3 micrograms/kg body weight merthiolate did not produce a damaging effect on the chromosomes of marrow, spleen and embryonal liver cells. Cyclophosphamide (in a dose of 25-50 micrograms/kg) used as positive control induced a significant rise in the frequency of chromosomal transformations.

Animals↗

Further efficacy studies on inactivated, aluminum-hydroxide-adsorbed vaccines against infectious coryza.

Specific-pathogen-free chickens were given infectious coryza vaccines containing inactivated cells of Haemophilus paragallinarum adsorbed onto an aluminum-hydroxide gel. The vaccines were protective when given subcutaneously or intramuscularly but not when given intranasally. Vaccines prepared with two different commercial brands of aluminum-hydroxide gel gave similar protection. Vaccines in which the inactivating agent was thimerosal were more protective than similar vaccines in which the inactivating agent was formalin. Monovalent vaccines protected against challenge only from organisms of the same agglutination serovar, whereas bivalent vaccines protected against challenge from organisms of either serovar. A single dose of the vaccine, given at 16 weeks of age, was not as effective as two doses given at 12 and 16 weeks of age in protecting against challenge 12, 25, or 56 weeks after vaccination.

Administration, Intranasal↗

Uncoupling the red cell sodium pump by proteolysis.

In situ proteolysis of Na,K-ATPase was studied using inside-out red cell membrane vesicles. Proteolysis of the enzyme in its "E1" conformation with either trypsin or chymotrypsin inactivated cation translocation more than ATP hydrolysis. This was evident both in the absence of intravesicular alkali cations when Na-ATPase was compared to ATP-dependent 22Na+ influx, and in the presence of K+ when Na+/K+ exchange was compared to (Na+ + K+)-activated ATPase. This differential loss in pump versus hydrolysis was observed also when the activities of only intact, non-leaky vesicles were compared and therefore reflects intramolecular uncoupling rather than nonspecific leakage. Although oligomycin and thimerosal, like trypsin and chymotrypsin, inhibit the enzyme's conformational step(s), neither effect uncoupling. It is concluded that specific cleavage(s) of Na,K-ATPase, at least as it exists in situ, alters the reaction sequence with respect to the normal ordered mechanism. Accordingly, cytoplasmic Na+ and extracellular K+ bind to the enzyme, stimulate phosphorylation (ATP + E1----E1P + ADP) and dephosphorylation (E2P----E2 + Pi), respectively, but each is then released to the same side from which it had bound; presumably release occurs prior to the conformational transitions of E1P to E2P and E2 to E1. This conclusion is supported by experiments showing that, ar micromolar ATP concentration, the hydrolytic activity (Na-ATPase) of the trypsinized but not the unmodified enzyme is stimulated by K+, consistent with earlier experiments (Hegyvary, C., and Post, R. L. (1971) J. Biol. Chem. 246, 5234-5240) showing that the K X E2 to K X E1 transition is slower than the E2 to E1 transition.

Biological Transport, Active↗

High degrees of astigmatism: are hydrogels the answer?

A bilateral compound hyperopic astigmat, with-the-rule, with incipient presbyopia and refractive amblyopia fitted with toric (custom) soft lenses in a monofit mode is described. Shortly after wearing the hydrogel toric lenses the patient developed a severe allergic reaction from a thimerosal based saline, which eventually resolved.

Adult↗

The effects of topical drugs and preservatives on the tears and corneal epithelium in dry eye.

Medications used in 'Dry Eye' patients are reviewed for their effects on the corneal surface including the overlying tear film. Preservatives are discussed, since they affect the properties of commercial preparations which may be instilled frequently as a substitute for normal tears. The major beneficial effect which a topically applied agent can have on epithelium is to supplement or stabilise the tear film. Thimerosal sometimes triggers a sensitivity reaction, and other mercurial compounds are unstable. Benzalkonium chloride compromises both corneal epithelium and tear film. Some cationic detergents, including chlorhexidine digluconate and polyquat, cause less disruption at prophylactic concentrations. The use of a small drop size is helpful in preventing toxic effects of preservatives. All preservatives should be avoided when unit doses of a sterile tear replacement, such as saline, can be made available. Topical antibiotics should be used only to control known bacterial infections, avoiding high concentrations of bacitracin, gentamicin, and neomycin. Steroids and antibiotic/steroid combinations must be used with great caution, and only when uncontrolled ocular inflammation justifies the risk of possible corneal ulceration.

Acetylcysteine↗

Chemical burns and skin preparation solutions.

Skin preparation burns associated with chemical agents are uncommon. They occur most frequently in those patients placed in the lithotomy position undergoing gynecologic operations, the burn being on the buttocks, and in those undergoing orthopedic operations, the burn being on the extremities and under a tourniquet. Thimerosal has been the most common agent to be implicated. The basic mechanism involves irritation coupled with maceration and pressure--conditions that can cause an acute pressure sore and superficial skin loss. To prevent these burns from occurring, the skin must not be abraded excessively before the final skin preparation solution is applied; the agent should not be allowed to pool and become trapped under the tourniquet or the torso of the patient, and the agent should be allowed to dry before the patient is draped. In our experience, the agent that has been the most effective and the least irritating to the skin during the time it has been used is povidone-iodine.

Adolescent↗

[Evaluation of the toxic action of prophylactic and therapeutic preparations on cell cultures of different types and origin. II. The cytotoxic action of adsorbed DPT vaccine and its components on cells of the continuous L132 line].

Different batches of the same preparation manufactured at the same enterprise, or at different enterprises, in accordance with the same manufacturing regulations have been found to be capable of producing a damaging effect of different intensity on the continuous cell culture L132. The titers vary, according to their cytotoxic effect, from 1 : 32 to 1 :2048. The components of B. pertussis antigens and thimerosal solutions have been found to produce the most pronounced cytotoxic effect on the cells. The comparison of the results of the titration of adsorbed DPT vaccine in cell cultures with clinical manifestations has shown correlation between a greater degree of cell damage in vitro and severe local reaction. Therefore, in the process of the quality control of preparations cell cultures provide more sensitive tests than laboratory animals, which is confirmed by our data obtained in revealing the toxic properties of adsorbed DPT vaccine and its components.

Adsorption↗

Determination of relative amounts of inositol trisphosphate receptor mRNA isoforms by ratio polymerase chain reaction.

The relative expression of different inositol 1,4,5-trisphosphate receptor (InsP3R) mRNA was determined in a selection of murine and rat cell types that are commonly used to study InsP3-mediated Ca2+ signaling. Different mRNA species (encoding the known InsP3R isoforms) were co-amplified using common polymerase chain reaction primer pairs that recognized sequences that are totally conserved between the various InsP3R. Specific identification of the co-amplified sequences was done by restriction site analysis. In cerebellum, mRNA encoding InsP3R-I accounted for > 90% of the total InsP3R mRNA. This isoform was also present in all other cell types tested and was often the major isoform. In contrast, the level of expression of the other isoforms was cell type-specific. A new InsP3R isoform (type V) was detected that had 94.5% sequence identity with the InsP3R-II in the amplified region. Interestingly, this isoform was largely expressed in murine but not in rat cells. We functionally characterized InsP3R-V using the mouse fibroblast C3H10T1/2 cells, where mRNA encoding InsP3R-V accounted for 76.4% of the total InsP3R mRNA. InsP3-induced Ca2+ release in permeabilized C3H10T1/2 cells was regulated by luminal and cytosolic Ca2+, stimulated by thimerosal, and inhibited by caffeine.

Animals↗

Inhibition of phosphodiesterase IV and intracellular calcium levels in human polymorphonuclear leukocytes.

Phosphodiesterase (PDE) isoenzyme type IV is the predominant cyclic AMP hydrolytic activity in polymorphonuclear leukocytes (PMNs). PDE IV inhibitors depress functional responses of PMNs but their influence on intracellular calcium concentration ([Ca2+]i) has not been extensively studied. The present study examined the effects of rolipram (a selective PDE IV inhibitor) on the chemotactic peptide formyl-methionyl-leucyl-phenylalanine (fMLP)-induced changes of [Ca2+]i in fura-2 loaded human PMNs. Rolipram (1 nM-10 microM) did not alter basal [Ca2+]i values. fMLP (10 nM approximately EC50) produced a transient calcium response, i.e., a peak followed by decay to a residual value above baseline. Peak [Ca2+]i values after fMLP were not altered but a faster decay and a lower residual [Ca2+]i were observed in rolipram (0.1-10 microM)-treated cells. fMLP added after thimerosal (20 microM) produced a peak followed by a sustained oscillatory response. Rolipram (up to 10 microM) did not alter the peak but inhibited the sustained response (-log IC50 = 6.39 +/- 0.12). The inhibitory effects of rolipram may be due to alterations in the mobilization of Ca2+ produced by the increase in the cellular content of cyclic AMP. SKF94120 (a selective PDE III inhibitor) produced minor effects on the fMLP-induced calcium response. SCA40 (a mixed PDE III/IV/V inhibitor) produced similar effects but was less potent than rolipram. Reduction of the calcium response probably underlies the inhibition of PMN functions produced by PDE IV inhibitors.

3',5'-Cyclic-AMP Phosphodiesterases↗

Electrophysiologic and morphologic effects of ophthalmic preparations on rabbit cornea epithelium.

The effects of several components of ophthalmic preparations on isolated rabbit cornea were studied by continuous electrophysiologic monitoring followed by fixation for scanning electron microscopy (SEM). Benzalkonium chloride (0.001 percent), thimerosal (0.0004 percent), and amphotericin B (0.0025 percent) all briefly increased ion transport, then greatly decreased epithelial resistance. Severe disruption of surface cell layers occurred simultaneously with resistance decrease. Silver nitrate (0.00017 percent) stimulated transport with less accompanying morphologic damage. Tetracaine (0.05 percent) disrupted epithelial function and caused exfoliation of several cell layers. Chlorobutanol (0.1 percent) produced a nearly complete loss of the squamous cell layer. Chloramphenicol, epinephrine, and pilocarpine produced minor changes in structure and electrophysiology at full clinical concentration. It was concluded that low concentrations of preservatives in ophthalmic preparations disrupt the barrier and transport properties of the corneal epithelium.

Amphotericin B↗

Growth and development of homograft tracheal transplants in the piglet model.

OBJECTIVE: To determine the growth characteristics of homograft tracheal transplants in piglets. DESIGN: Prospective controlled animal study. SETTING: Clinical animal laboratory. SUBJECTS: Seventeen Yorkshire swine piglets. INTERVENTIONS: The tracheae of adult Yorkshire swine were harvested and treated with formaldehyde, thimerosal, and acetone to remove immunogenic major histocompatibility complexes. Eleven piglets had these chemically treated homografts transplanted into 6-cm surgically created tracheal defects. The transplants were stented. Three control piglets had a 6-cm anterior tracheofissure, no transplant, and surgical placement of the stent. Three other control piglets had no transplant, and the stent was placed endoscopically. MAIN OUTCOME MEASURES: Growth outcome measurements were tracheal length and diameter. Functional outcome measurements were lumen patency and graft viability indicated by cartilage retention. RESULTS: The mean diameter of the tracheae in the stented tracheal transplant group was 11.7 mm before transplantation and 6.6 mm 2 months after transplantation. The transplanted segments were significantly malacic 2 months after transplantation. The mean diameter of the tracheae in the tracheofissure group was 9.0 mm before surgery and 11.0 mm 2 months after surgery. The mean diameter of the tracheae in the endoscopically stented group was 11.0 mm before surgery and 14.0 mm 2 months after stent placement. All homografts showed evidence of extensive resorption of the graft cartilage. The graft cartilage was replaced by collagen, with minimal evidence of neochondrification. There was no evidence of host-vs-graft rejection. All grafted trachea had severe tracheomalacia with granulation tissue. CONCLUSIONS: Homograft tracheal transplantation results in a tracheal segment that is replaced with collagen. The transplanted cartilage is resorbed, leaving a significantly malacic segment. Homograft tracheal transplantation might result in a small malacic airway with little potential for growth when performed in children.

Animals↗