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PubMed2004

A comparison of the effects of ocular preservatives on mammalian and microbial ATP and glutathione levels.

The aim of this study was to investigate the mechanism of action of the preservative sodium chlorite (NaClO2), and the relationship with intracellular glutathione depletion. A detailed comparison of the dose responses of two cultured ocular epithelial cell types and four species of microorganism was carried out, and comparisons were also made with the quaternary ammonium compound benzalkonium chloride (BAK), and the oxidant hydrogen peroxide (H2O2). The viability of mammalian and microbial cells was assessed in the same way, by the measurement of intracellular ATP using a bioluminescence method. Intracellular total glutathione was measured by reaction with 5,5'-dithiobis-2-nitrobenzoic acid in a glutathione reductase-dependent recycling assay. BAK and H2O2 caused complete toxicity to conjunctival and corneal epithelial cells at approximately 25 ppm, in contrast to NaClO2, where > 100 ppm was required. The fungi Candida albicans and Alternaria alternata had a higher resistance to NaClO2 than the bacteria Staphyloccus aureus and Pseudomonas aeruginosa, but the bacteria were extremely resistant to H2O2. NaClO2 caused substantial depletion of intracellular glutathione in all cell types, at concentrations ranging from < 10 ppm in Pseudomonas, 25-100 ppm in epithelial cells, to > 500 ppm in fungal cells. The mechanisms of cytotoxicity of NaClO2, H2O2 and BAK all appeared to differ. NaClO2 was found to have the best balance of high antibacterial toxicity with low ocular toxicity. The lower toxicity of NaClO2 to the ocular cells, compared with BAK and H2O2, is in agreement with fewer reported adverse effects of application in the eye.

Adenosine Triphosphate↗
PubMed2001

Effects of daily dermal application of DEET and epermethrin, alone and in combination, on sensorimotor performance, blood-brain barrier, and blood-testis barrier in rats.

DEET and permethrin were implicated in the development of illnesses in some veterans of the Persian Gulf War. This study was designed to investigate the effects of daily dermal application of these chemicals, alone or in combination, on the permeability of the blood-brain barrier (BBB) and blood-testes barrier (BTB) and on sensorimotor performance in male Sprague-Dawley rats. Groups of five rats were treated with a dermal daily dose of 4, 40, or 400 mg/kg DEET in ethanol or 0.013, 0.13, or 1.3 mg/kg permethrin in ethanol for 60 d. A group of 10 rats received a daily dermal dose of ethanol and served as controls. BBB permeability was assessed by injection of an iv dose of the quaternary ammonium compound [3H]hexamethonium iodide. While permethrin produced no effect on BBB permeability, DEET alone caused a decrease in BBB permeability in brainstem. A combination of DEET and permethrin significantly decreased the BBB permeability in the cortex. BTB permeability was decreased by treatment with DEET alone and in combination with permethrin. The same animals underwent a battery of functional behavior tests 30, 45, and 60 d after exposure to evaluate their sensorimotor abilities. All treatments caused a significant decline in sensorimotor performance in a dose- and time-dependent manner. These results show that daily dermal exposure to DEET, alone or in combination with permethrin, decreased BBB permeability in certain brain regions, and impaired sensorimotor performance.

Administration, Topical↗
PubMed1956

Effect of silicate on Gram staining and viability of pneumococci and other bacteria.

Application of silicate solutions to living or heat-killed pneumococci and to certain "viridans" streptococci causes their conversion from a Gram-positive to a Gram-negative state. The original staining properties can be restored by suspending the silicate-treated bacteria in alkaline solutions of various salts but not by simple washing in water. Living pneumococci and the strains of streptococci whose staining properties are similarly affected are killed when suspended in silicate solutions. In other Gram-positive species silicate causes conversion to Gram negativity but restoration to positivity occurs upon washing in water. In a third group of Gram-positive organisms silicate has no effect on the Gram reaction. The viability of organisms in these two groups is unaffected by silicate under the conditions employed. No effect on staining or viability of Gram-negative bacteria has been observed. The effects of silicate on staining and viability are inhibited by nutrient broth or whole serum but not by purified serum albumin. Lecithin, choline, and other substituted ammonium compounds also inhibit the effects of silicate on pneumococci.

Bacteria↗
PubMed2006

Use of germicides in the home and the healthcare setting: is there a relationship between germicide use and antibiotic resistance?

BACKGROUND: The spread of antibiotic-resistant pathogens represents an increasing threat in healthcare facilities. Concern has been expressed that the use of surface disinfectants and antiseptics may select for antibiotic-resistant pathogens. OBJECTIVE: To review the scientific literature on whether there is a link between use of germicides (ie, disinfectants and antiseptics) and bacterial resistance to antibiotics. In addition, we will review whether antibiotic-resistant bacteria exhibit altered susceptibility to germicides that are recommended for use as disinfectants or antiseptics. DESIGN: A review of the appropriate scientific literature. RESULTS: In the laboratory, it has been possible to develop bacterial mutants with reduced susceptibility to disinfectants and antiseptics that also demonstrate decreased susceptibility to antibiotics. However, the antibiotic resistance described was not clinically relevant because the test organism was rarely a human pathogen, the altered level of antimicrobial susceptibility was within achievable serum levels for the antibiotic, or the antibiotic tested was not clinically used to treat the study pathogen. Similarly, wild-type strains with reduced susceptibility to disinfectants (principally, quaternary ammonium compounds) and antiseptics (principally, triclosan) have been reported. However, because the concentration of disinfectants used in the healthcare setting greatly exceeds the concentration required to kill strains with reduced susceptibility to disinfectants, the clinical relevance of these observations is questionable. CONCLUSION: To date, there is no evidence that using recommended antiseptics or disinfectants selects for antibiotic-resistant organisms in nature. Disinfectants and antiseptics should be used when there are scientific studies demonstrating benefit or when there is a strong theoretical rationale for using germicides.

Anti-Bacterial Agents↗
PubMed1996

Disinfection and sterilization of patient-care items.

This article provides recommendations on the preferred method for disinfection and sterilization of patient-care items based on the intended use of the item (i.e., critical, semicritical, noncritical). The chemical disinfectants recommended for patient-care items and instruments include glutaraldehyde, hydrogen peroxide, peracetic acid, sodium hypochlorite, alcohol, iodophors, phenolics, and quaternary ammonium compounds. The choice of disinfectant, concentration, and exposure time is based on the risk of infection associated with the use of the item. The sterilization methods briefly discussed include steam sterilization, ethylene oxide, dry heat, and the new low-temperature sterilization technologies. When properly used, these disinfection and sterilization processes can ensure the safe use of invasive and noninvasive medical devices. However, this requires strict adherence to current cleaning, disinfection, and sterilization guidelines.

Cross Infection↗
PubMed1997

Activity of disinfectants against vancomycin-resistant Enterococcus faecium.

Vancomycin-resistant enterococci (VRE) often contaminate the hospital environment. We examined the activity of commonly used disinfectants against eight strains of VRE, using a quantitative suspension test method. Isopropyl alcohol and sodium hypochlorite were highly effective. Hydrogen peroxide was ineffective for all strains. After 10 minutes of incubation (the manufactures' recommended time of exposure), three phenolic and three quaternary ammonium compounds also were highly effective. After 3 minutes of exposure, however, occasional failures did occur. With the exception of 3% hydrogen peroxide, most disinfectants appear to be active against VRE.

Anti-Bacterial Agents↗
PubMed2001

Disinfectant and antibiotic resistance of lactic acid bacteria isolated from the food industry.

Quaternary ammonium compounds (QACs) are widely used as disinfectant in medical and food environments. There is a growing concern about the increasing incidence of disinfectant-resistant microorganisms from food. Disinfectant-resistant lactic acid bacteria (LAB) may survive disinfection and cause spoilage problems. Moreover, resistant LAB may potentially act as a reservoir for resistance genes. A total number of 320 LAB from food industry and meat were screened for resistance to the QAC benzalkonium chloride (BC). Out of 320 strains, five strains (1.5%) were considered to be resistant and 56 (17.5%) were tolerant to BC. The resistant strains were isolated from food processing equipment after disinfection. The resistant, tolerant, and some sensitive control bacteria were examined for susceptibility to 18 different antibiotics, disinfectants, and dyes using disc agar diffusion test and microdilution method. Little systematic cross-resistance between BC and any of the antimicrobial agents tested were detected except for gentamycin and chlorhexidine. A BC-tolerant strain was much easier to adapt to higher levels of BC as compared to a BC-sensitive strain. No known gram-positive QAC resistance genes (qacA/B, qacC, qacG, and qacH) were detected in the BC-resistant strains. Identification to species level of the BC-resistant isolates was carried out by comparative analysis of 16S-rDNA sequencing. In conclusion, resistance to BC is not frequent in LAB isolated from food and food environments. Resistance may occur after exposure to BC. The BC resistant isolates showed no cross-resistance with other antimicrobial compounds, except for gentamycin and chlorhexidine. Nevertheless, BC-resistant LAB may be isolated after disinfection and may contribute to the dissemination of resistance.

Anti-Infective Agents, Local↗
PubMed2006

Accumulation of glycinebetaine in rice plants that overexpress choline monooxygenase from spinach and evaluation of their tolerance to abiotic stress.

BACKGROUND AND AIMS: Glycinebetaine (GB), a quaternary ammonium compound, is a very effective compatible solute. In higher plants, GB is synthesized from choline (Cho) via betaine aldehyde (BA). The first and second steps in the biosynthesis of GB are catalysed by choline monooxygenase (CMO) and by betaine aldehyde dehydrogenase (BADH), respectively. Rice (Oryza sativa), which has two genes for BADH, does not accumulate GB because it lacks a functional gene for CMO. Rice plants accumulate GB in the presence of exogenously applied BA, which leads to the development of a significant tolerance to salt, cold and heat stress. The goal in this study was to evaluate and to discuss the effects of endogenously accumulated GB in rice. METHODS: Transgenic rice plants that overexpressed a gene for CMO from spinach (Spinacia oleracea) were produced by Agrobacterium-mediated transformation. After Southern and western blotting analysis, GB in rice leaves was quantified by (1)H-NMR spectroscopy and the tolerance of GB-accumulating plants to abiotic stress was investigated. KEY RESULTS: Transgenic plants that had a single copy of the transgene and expressed spinach CMO accumulated GB at the level of 0.29-0.43 micromol g(-1) d. wt and had enhanced tolerance to salt stress and temperature stress in the seedling stage. CONCLUSIONS: In the CMO-expressing rice plants, the localization of spinach CMO and of endogenous BADHs might be different and/or the catalytic activity of spinach CMO in rice plants might be lower than it is in spinach. These possibilities might explain the low levels of GB in the transgenic rice plants. It was concluded that CMO-expressing rice plants were not effective for accumulation of GB and improvement of productivity.

Betaine↗
PubMed2002

Pseudomonas aeruginosa cells adapted to benzalkonium chloride show resistance to other membrane-active agents but not to clinically relevant antibiotics.

Our objective was to determine whether strains of Pseudomonas aeruginosa can adapt to growth in increasing concentrations of the disinfectant benzalkonium chloride (BKC), and whether co-resistance to clinically relevant antimicrobial agents occurs. Attempts were made to determine what phenotypic alterations accompanied resistance and whether these explained the mechanism of resistance. Strains were serially passaged in increasing concentrations of BKC in static nutrient broth cultures. Serotyping and genotyping were used to determine purity of the cultures. Two strains were examined for cross-resistance to other disinfectants and antibiotics by broth dilution MIC determination. Alterations in outer membrane proteins and lipopolysaccharide (LPS) expressed were examined by SDS-PAGE. Cell surface hydrophobicity and charge, uptake of disinfectant and proportion of specific fatty acid content of outer and cytoplasmic membranes were determined. Two P. aeruginosa strains showed a stable increase in resistance to BKC. Co-resistance to other quaternary ammonium compounds was observed in both strains; chloramphenicol and polymyxin B resistance were observed in one and a reduction in resistance to tobramycin observed in the other. However, no increased resistance to other biocides (chlorhexidine, triclosan, thymol) or antibiotics (ceftazidime, imipenem, ciprofloxacin, tobramycin) was detected. Characteristics accompanying resistance included alterations in outer membrane proteins, uptake of BKC, cell surface charge and hydrophobicity, and fatty acid content of the cytoplasmic membrane, although no evidence was found for alterations in LPS. Each of the two strains had different alterations in phenotype, indicating that such adaptation is unique to each strain of P. aeruginosa and does not result from a single mechanism shared by the whole species.

Anti-Bacterial Agents↗
PubMed2025

Genomic structure of class 1 and 2 integrons in non-typhoidal Salmonella isolated from food animals and related meat products in the USA.

OBJECTIVES: Integrons facilitate the capture and expression of exogenous genes, including antimicrobial resistance (AMR) genes. This study aimed to detect the presence of integrons, examine their genomic structure and location, and analyse integron-associated AMR, virulence and stress response genes in Salmonella using WGS. METHODS: WGS data from 193 Salmonella strains, representing 38 serotypes isolated from food animals and related meat products (2001-2019), were analysed using bioinformatic tools to assess integron presence and characterize their genomic architectures. RESULTS: Of 193 isolates, 116 (60.1%) harboured class 1 and/or class 2 integrons. Class 1 integrons alone were detected in 105 isolates, with some containing multiple copies. One S. Infantis isolate harboured only class 2 integrons, whereas 10 others contained both classes. No class 3-5 integrons were found. Twenty-seven class 1 integrons were chromosomal; the rest were plasmid-associated, linked to various plasmid incompatibility (Inc) types. Sixty-nine distinct AMR genes conferring resistance to 11 antimicrobial classes were found in integron cassettes or integron-associated plasmids. Genes linked to resistance to quaternary ammonium compounds and heavy metals, as well as ISs and transposons, were also identified. Significant virulence and stress response genes and proteins such as groES-groEL, LysR and EAL (glutamate, alanine and leucine) were common in integron cassettes. CONCLUSIONS: Class 1 integrons are prevalent in MDR Salmonella isolates from food animals and related meat products and are linked to diverse plasmid types. Their association with AMR, virulence and stress response genes underscores their role in AMR dissemination, and bacterial adaptation and pathogenicity.

Integrons↗
PubMedDate not supplied

Extraction, identification and quantitation of succinylcholine in embalmed tissue.

Succinylcholine, a bis-quaternary ammonium compound, was extracted from embalmed tissues as an ion-pair with hexanitrodiphenylamine in methylene chloride. The evaporated ion-pair residue is demethylated with sodium benzenethiolate. The tertiary amine formed is identified and quantitated by gas chromatography/mass spectrometry (GC/MS) utilizing a 25 meter glass capillary column coated with SE 52. Identification is accomplished by retention time and mass spectrometry. Quantitation is performed after the addition of deuterated succinylcholine as an internal standard by focusing the mass spectrometer on m/z 58 (for demethylated succinylcholine) and m/z 62 (for the internal standard). The method is applied to the quantitation of succinylcholine from the embalmed kidney, liver, and muscle of rats injected i.m. with 10-200 mg/kg. After six months of storage, the succinylcholine can still be identified and quantitated with highest concentrations found in the muscle injection site. Concentrations as low as 5 ng/g are easily detected.

Animals↗
PubMedDate not supplied

Determination of succinylcholine in tissues by TLC, GC/NPD, and GC/MS.

Succinylcholine, a bis-quaternary ammonium compound, is considered an elusive analyte due to rapid hydrolysis by pseudocholinesterase in plasma. However, tissue acetylcholinesterase is relatively inactive toward this substrate. Hence, analysis of tissues of succinylcholine-treated animals may reveal the presence of unchanged drug. This report describes three chromatographic techniques (TLC, GC/NPD, and GC/MS) which may be applied to the analysis of this substance in human tissues. Validation of the techniques was accomplished using tissues from rats treated with succinylcholine.

Animals↗
PubMed2003

Choline uptake in human intestinal Caco-2 cells is carrier-mediated.

The objective of the current investigation was to examine the transport characteristics of choline, an endogenous quaternary ammonium compound, into human intestinal Caco-2 cells; the transport of choline has not been characterized in human intestine. The cellular accumulation of choline was independent of an inwardly directed Na(+) gradient and demonstrated temperature dependence and saturability. Using the initial uptake rates, choline accumulation was best characterized by a Michaelis-Menten equation and a diffusion component with a K(m) and V(max) of 110 +/- 3 micro mol/L and 2800 +/- 250 pmol/(mg protein. 10 min), respectively. Choline uptake was significantly inhibited by an excess of choline itself and by hemicholinium-3, a structural analog of choline. However other hydrophilic organic cations, such as tetraethylammonium (TEA) and N-methylnicotinamide (NMN), did not affect choline uptake in Caco-2 cells. Additionally, two typical p-glycoprotein substrates, daunomycin and verapamil, both inhibited choline accumulation. However the opposite was not true: choline did not inhibit DNM accumulation in Caco-2 cells. These results indicate the presence of a carrier-mediated transport system for choline in Caco-2 cells. The substrate specificity of this carrier is unlike that seen in the rat intestinal epithelium, and the human transport protein is distinct from those for TEA and NMN. P-glycoprotein substrates may inhibit choline uptake through specific or nonspecific interactions with the choline transporter.

Caco-2 Cells↗
PubMed1986

Risk factors for wound infections after genitourinary reconstructive surgery.

In January through August 1983, eight children developed wound infections after they underwent genitourinary reconstructive surgery at one hospital. Cultures of purulent drainage yielded Pseudomonas aeruginosa in two cases, Streptococcus faecalis in one case, both S. faecalis and Staphylococcus epidermidis in one case, and S. faecalis and other Gram-negative rods in four cases. A study in which uninfected children who had had genitourinary reconstructive surgery were used as controls revealed that all cases and controls had received cephalosporin prophylaxis. Antibiograms revealed that all organisms were resistant to the cephalosporin antibiotics tested. Risk of infection was associated with shaving (p less than 0.001, Fisher's exact test) and exposure to one nurse (nurse X) (p = 0.003, Fisher's exact test) and one surgeon (surgeon X) (p = 0.007, Fisher's exact test). Nurse X was the private scrub nurse for surgeon X; she was present only at operations surgeon X performed (p less than 0.001, Fisher's exact test) and had no reported or observed practices different from those of other surgical nurses. However, surgeon X had two practices that were notably different from those of other surgeons: 1) he used quaternary ammonium compounds, acetone/alcohol, and 1 per cent hexachlorophene for preoperative skin preparation; and 2) he prepared the operative site himself. In addition, he was observed to prepare the operative site by beginning centrally, moving peripherally, and returning to the central area with the same sponge. These findings suggested that the shaving of the operative site and the use of certain agents and/or different preparatory techniques may have contributed to increased risk of wound infections. These wound infections were caused by organisms resistant to the prophylactic antibiotic(s) used.

Anti-Bacterial Agents↗
PubMed1998

Longitudinal changes in lung function associated with aspects of swine-confinement exposure.

Several aspects of swine-confinement farming appear to be leading to adverse respiratory effects. This study was set up in a longitudinal design to study the association between certain characteristics of farms or the way they are run and a decline in lung function. A cohort of 171 pig farmers was observed for 3 years. Lung function was measured. Exposure to farm characteristics was determined at the start of the observation period, using data from standardized farm surveys and from diaries kept by the participants. Mean decline in lung function was 73 mL/year for forced expiratory volume in 1 second (FEV1) and 55 mL/year for forced vital capacity (FVC). A longitudinal decline in FEV1 was significantly associated with the use of quaternary ammonium compounds as disinfectants (an additional 43 mL/year) and also with the use of an automated dry feeding system (an additional 28 mL/year). The association with the use of wood shavings as bedding material was not statistically significant. The impact of these characteristics in a longitudinal study provides stronger evidence for causal inference than that shown in previous cross-sectional designs. This may be useful in promoting preventive measures.

Adult↗
PubMed2003

Production of the novel mesangial autocrine growth factors GDNF and IL-10 is regulated by the immunomodulator AS101.

Mesangial cell (MC) proliferation is essential for the pathogenesis and progression of various glomerular diseases. This study shows that glial cell line-derived neurotrophic factor (GDNF) and IL-10 are mesangial autocrine growth factors that play a pivotal role in rat MC proliferation in vitro. Downstream targets of GDNF signaling and their role in MC hyperplasia are identified. The phosphatidylinositol 3-kinase (PI3K) pathway and its downstream target NF-kappaB were found to mediate GDNF-induced MC mitogenesis. This pathway also mediates GDNF-induced decrease in the cyclin-dependent kinase inhibitor p27(kip1) expression, resulting in the increased formation of cyclin D1/cdk4 and cyclin E/cdk2 complexes, followed by hyperphosphorylation of retinoblastoma, a key event for G1 to S phase progression. IL-10 appears to be a more potent MC growth factor that negatively regulates GDNF expression. Indeed, its inhibition by the nontoxic tellurium anti-IL-10 compound, ammonium trichloro(dioxoethylene-o,o') tellurate (AS101), extensively decreased MC clonogenicity despite GDNF upregulation. Identification of the mesangial GDNF and IL-10 pathways as critical mediators of mesangial cell proliferation may provide another target for therapeutic intervention in certain glomerular diseases. In vivo animal studies using AS101, currently undergoing phase II clinical trials on cancer patients, are warranted to determine its potential in the management of glomerular diseases associated with mesangial cell proliferation.

Adjuvants, Immunologic↗
PubMed1978

Fine structure, physiology and biochemistry of arthrospore germination in Streptomyces antibioticus.

During germination, Streptomyces antibioticus arthrospores passed through stages: darkening, swelling and germ tube emergence. The first stage, darkening, whose main features were a decrease in absorbance and a loss of refractility, only required exogenous divalent cations (Ca2+, Mg2+ or Fe2+) and energy that can be obtained from the spore reserves. This stage was blocked by agents that inhibit ATP formation but not by antibiotics that inhibit macromolecular synthesis. The second stage, swelling, needed an exogenous carbon source and was not blocked by mitomycin C. In this stage, the spores exhibited the highest cytochrome oxidase and catalase activities and respiratory quotient. The last stage, germ tube emergence, required additional carbon and nitrogen sources. Ammonium compounds were superior to nitrate. Dry weight remained constant during the stages of darkening and swelling, with a rapid increase from the moment of germ tube emergence. Optimum pH and temperature for germination were 8.0 and 45 degrees C, respectively. Heat treatment (55 degrees C for 10 min) had no effect on germination. The fine structure of the spore underwent important changes during germination. The wall of the swollen spore became stratified and the inner layer was continuous with the germ tube wall. Macromolecular synthesis occurred in the sequence RNA, protein and then DNA. Rifampicin, streptomycin and mitomycin C prevented synthesis when added at the start of incubation. The same effect was obtained if the addition was made during germination, except with mitomycin C which inhibited DNA, but not RNA and protein synthesis.

Anti-Bacterial Agents↗
PubMed1989

Physical and biochemical characterization of the qacA gene encoding antiseptic and disinfectant resistance in Staphylococcus aureus.

We have previously cloned a 3.5 kb fragment from the Staphylococcus aureus multiresistance plasmid pSK1 which carries the qacA determinant responsible for linked resistance to acriflavine (Acr), ethidium bromide (Ebr), quaternary ammonium compounds (Qar), propamidine isethionate (Pir), and diamidinodiphenylamine dihydrochloride (Ddr). This report presents a biochemical and physical analysis of qacA and shows the widespread carriage of this gene on S. aureus resistance plasmids. Tn5 insertion mutagenesis defined the extent of qacA to within 2.40 kb of pSK1 DNA. Examination of the expression of insertion and deletion mutants of the cloned qacA sequences in both maxicells and minicells led to the association of a 50 kDa protein, designated QacA, with the AcrEbrQarPirDdr phenotype. Based on fluorimetric and isotopic assays used to determine the extent of accumulation of ethidium bromide by S. aureus strains harbouring pSK1, we propose that the basis of AcrEbrQarPirDdr in S. aureus is a qacA-mediated efflux system.

Anti-Infective Agents, Local↗