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Antibiotic resistance in Staphylococcus aureus isolated at an Australian hospital between 1946 and 1981.

A total of 517 strains of Staphylococcus aureus isolated at a hospital in Melbourne, Australia between 1946 and 1981 was examined for resistance to a range of antimicrobial agents and for the presence of plasmid DNA. The use of mixed-culture transfer and restriction endonuclease analysis showed that the determinants for resistance to penicillin and to the heavy metals were carried by several related plasmids of (15-23) X 10(6) mol. wt, and that tetracycline resistance was encoded on a plasmid of 2.8 X 10(6) mol. wt in strains isolated before 1970. These phenotypes were chromosomally encoded in the majority of strains isolated thereafter. Resistance to chloramphenicol throughout the study period was plasmid-mediated. Of five aminoglycoside-resistance phenotypes, one was plasmid-mediated and three were chromosomally encoded. The remaining phenotype, specifying low-level gentamicin resistance, was found to be located on the chromosome of early isolates, but in later strains was borne by an 18 X 10(6) mol. wt plasmid which also encoded resistance to quaternary ammonium compounds.

Aminoglycosides↗

Inhibition of Gibberellin Production in the Fungi Gibberella fujikuroi and Sphaceloma manihoticola by Plant Growth Retardants.

The effect of different types of plant growth retardants on fungal gibberellin (GA) formation has been studied in cultures of Gibberella fujikuroi and Sphaceloma manihoticola. Quaternary ammonium compounds (chlormequat chloride, mepiquat chloride, Amo-1618), triazoles (uniconazole and several experimental compounds), and the norbornanodiazetine tetcyclacis inhibited GA biosynthesis in both fungal species. Concentrations between 2 x 10(-4) and 10(-9)m were required for a 50% inhibition of the production of gibberellin A(3) in Gibberella fujikuroi and of giberellin A(4) in Sphaceloma manihoticola. The formation of other prominent GAs was affected at a similar degree of intensity. Tetcyclacis was the most active compound in both fungi. Compared to the growth retardants mentioned above, the biological activity of chlorphonium chloride was low. The acylcyclohexanediones prohexadione and LAB 198 999 had virtually no activity. Most likely, this lack of activity is due to a rapid metabolism of the compounds in the cultures. For the triazole-type compounds and tetcyclacis, a relatively distinct correlation exists in their ability to inhibit GA formation in fungal cultures, to block ent-kaurene oxygenase in a cell-free system, and to reduce shoot growth of rice seedlings. Due to differences in their metabolic fate and species specificities, such conclusions cannot be made for the other compounds.

Journal Article↗

Responses of halophytes to high salinities and low water potentials.

The effects of nonsaline polyethylene glycol (PEG)-6000 and saline seawater solutions of comparable osmotic potential on the concentrations of organic solutes and inorganic ions in the tissues of halophytes (Plantago maritima L., Triglochin maritima L., Limonium vulgare Mill., Halimione portulacoides (L.) Aell) have been investigated. Studies were made to determine whether high salinities induce specific ion effects that are absent in plants grown in nonsaline solutions of comparable osmotic potential. Over-all, the responses of each species to the two different treatments (seawater or PEG) are similar; the accumulation of organic solutes (compatible osmotica) in tissues is primarily correlated with a decrease in the osmotic potential of culture solutions. Depending on the species, sorbitol, proline, reducing sugars, quaternary ammonium compounds, and alpha-amino nitrogen accumulate in tissues as the water potential of the tissues falls. Within a species there are differences in the concentrations of inorganic ions and organic solutes between roots and shoots of plants grown at high salinities or at high concentrations of PEG.

Journal Article↗

Ion Homeostasis in Chloroplasts under Salinity and Mineral Deficiency: II. Solute Distribution between Chloroplasts and Extrachloroplastic Space under Excess or Deficiency of Sulfate, Phosphate, or Magnesium.

Spinach (Spinacia oleracea var "Yates") plants grown hydroponically were exposed to an excess or deficiency of various mineral ions. Solutes were measured in leaf extracts and in isolated intact chloroplasts. Under phosphate (120 millimoles per liter NaH(2) PO(4)), sulfate (200 millimolar per liter (Na(2) SO(4)), or magnesium excess (150 millimolar per liter MgCl(2)), concentrations of these ions in leaf extracts increased, but in chloroplasts, concentrations of all ions remained constant. Concentrations of quarternary ammonium compounds in chloroplasts increased. Under mild phosphate or magnesium deficiency, concentrations of these ions decreased in chloroplasts less than in whole leaf extracts. Under severe sulfate deficiency causing chlorosis in younger leaves, sulfate concentrations in chloroplasts remained even unchanged, despite a drastic decrease of sulfate concentrations both in green and in chlorotic leaves. Together with results from a companion study (G Schröppel-Meier, WM Kaiser 1988 Plant Physiol 87: 822-827) our data demonstrate that leaf cells are able to keep the concentrations of several mineral ions rather constant in metabolically active compartments even at extremely large variations of ion concentrations in the culture solution and in the leaves.

Journal Article↗

Comparative Physiological Evidence that beta-Alanine Betaine and Choline-O-Sulfate Act as Compatible Osmolytes in Halophytic Limonium Species.

The quaternary ammonium compounds accumulated in saline conditions by five salt-tolerant species of Limonium (Plumbaginaceae) were analyzed by fast atom bombardment mass spectrometry. Three species accumulated beta-alanine betaine and choline-O-sulfate; the others accumulated glycine betaine and choline-O-sulfate. Three lines of evidence indicated that beta-alanine betaine and choline-O-sulfate replace glycine betaine as osmo-regulatory solutes. First, tests with bacteria showed that beta-alanine betaine and choline-O-sulfate have osmoprotective properties comparable to glycine betaine. Second, when beta-alanine betaine and glycine betaine accumulators were salinized, the levels of their respective betaines, plus that of choline-O-sulfate, were closely correlated with leaf solute potential. Third, substitution of sulfate for chloride salinity caused an increase in the level of choline-O-sulfate and a matching decrease in glycine betaine level. Experiments with (14)C-labeled precursors established that beta-alanine betaine accumulators did not synthesize glycine betaine and vice versa. These experiments also showed that beta-alanine betaine synthesis occurs in roots as well as leaves of beta-alanine betaine accumulators and that choline-O-sulfate and glycine betaine share choline as a precursor. Unlike glycine betaine, beta-alanine betaine synthesis cannot interfere with conjugation of sulfate to choline by competing for choline and does not require oxygen. These features of beta-alanine betaine may be advantageous in sulfate-rich salt marsh environments.

Journal Article↗

Free radical reduction by thioredoxin reductase at the surface of normal and vitiliginous human keratinocytes.

Cell cultures of human keratinocytes contain membrane-associated thioredoxin reductase that is extremely active in reducing radicals on the outer plasma membrane. This enzyme activity was confirmed by its purification from cultures of stratified human keratinocytes by affinity column chromatography. The enzyme was assayed both in vivo and in vitro using a spin-labeled quaternary ammonium compound as the substrate, under saturating conditions in free radical substrate. Specific activities were determined by monitoring the sequential decrease in the amplitude of the electron spin resonance signal per unit of cell protein. The following properties were found: Cultures of adult stratified cells have approximately twice the thioredoxin reductase activity of neonatal cells. The enzyme is inhibited by thioprotein inhibitors (i.e., parachloromecuribenzoate and dinitrochlorobenzene). The activity is regulated by calcium concentrations of the cell culture medium. Stratified keratinocytes are half as active in medium containing 2 mM Ca++ compared with 0.1 mM Ca++ concentration. Product inhibition of the enzyme occurs with oxidized coenzyme NADP+ (i.e., 87% inhibition of enzyme activity over 30 min). The enzyme is heat stable at temperatures of 70 degrees C for 10 min. It is inactivated at 75 degrees C. A comparative study of thioredoxin reductase activity on stratified differentiated and undifferentiated rapidly growing cells was performed. Also, enzyme activity was quantitated for cultured keratinocytes isolated from vitiliginous and normal skin of the same donor. The results of this study, and the connection between this enzyme activity and UV-generated free radicals are reconciled in terms of the mechanism of action and metabolic activity of thioredoxin reductase.

Adult↗

Two different mechanisms are involved in the extremely high-level benzalkonium chloride resistance of a Pseudomonas fluorescens strain.

A Pseudomonas fluorescens strain, PFRB, which we previously isolated as a contaminant in a batch of benzalkonium chloride (BAC) stock solution, exhibits high-level resistance, not only to BAC, but also to other cationic surfactants belonging to disinfectants classified as quaternary ammonium compounds (QACs). In this study, we analyzed the resistance mechanism of the strain to BAC and other disinfectants. We obtained results suggesting that two different mechanisms, reduced adsorption of BAC to the cell surface and an energy-dependent mechanism which is most probably an efflux system, were implicated in the high-level resistance to BAC. Reduced adsorption of BAC is likely due to the decreased negative cell surface charge of the strain. The putative efflux system seems to be unique in that it excretes only a certain range of cationic membrane-acting disinfectants belonging to QACs.

Adsorption↗

Allergy to suxamethonium: persisting abnormalities in skin tests, specific IgE antibodies and leucocyte histamine release.

Twenty-one patients, who had previously experienced an anaphylactic reaction to suxamethonium during general anaesthesia, were selected for this study. Initially, skin tests with muscle relaxants were carried out in the twenty-one patients, detection of specific anti-choline IgE in nineteen, and leucocyte histamine release in seventeen. These three tests were then repeated between 1 year and 4 years after the initial evaluation. In the majority of patients, sensitization to the muscle relaxants persisted for more than 1 year after the anaphylactic reaction. Only three patients out of twenty-one (4%) had negative skin tests when retested 1-4 years later. A reduction in leucocyte histamine release was noticed in one of the seventeen retested patients (6%). Modifications of anti-choline IgE were observed in five of nineteen patients (26%). The persistence of sensitization to suxamethonium may result from repeated stimulation by occasional contacts with quaternary ammonium compounds. This study demonstrates the reliability of skin tests, leucocyte histamine release and detection of anti-choline IgE to diagnose allergic reactions to suxamethonium, even when they are performed a long time after the initial anaphylactic reaction.

Alcuronium↗

A simple most probable number method for the enumeration of sulphate-reducing bacteria in biocide containing waters.

A simple most probable number (MPN) method has been developed for the enumeration of sulphate-reducing bacteria (SRB) in biocide-containing waters. The medium used is based on source water, it contains no toxic thioglycollate and is resistant to oxidation through mishandling. Reduction is by a suspension of Pseudomonas putida which acts as a powerful adsorbent of biguanide, phenolic, quaternary ammonium compound, glutaraldehyde and isothiazolone biocides. Good recoveries of SRB type strains were obtained using this method and were comparable to other published techniques. Recovery of SRB in mixed culture was comparable to that using a standard laboratory technique.

Adsorption↗

Sensitivity of methicillin-resistant Staphylococcus aureus strains to some antibiotics, antiseptics and disinfectants.

The effects of antibiotics, antiseptics and disinfectants against some methicillin-resistant (MRSA) and methicillin-sensitive (MSSA) Staphylococcus aureus strains have been studied. The MRSA and MSSA strains were equally sensitive to phenols, esters of para(4)-hydroxybenzoic acid and chlorhexidine but MRSA strains were slightly more resistant to quaternary ammonium compounds and considerably more so to dibromopropamidine isothionate. Some MRSA strains were also resistant to phenylmercuric nitrate (but not another organomercurial, thiomersal), mercuric chloride and cadmium chloride. All MRSA strains produced beta-lactamase. Strains from the Royal Free Hospital, London were highly resistant to beta-lactam antibiotics, erythromycin, trimethoprim and tetracyclines but were sensitive to other antibiotics. One strain from the University Hospital of Wales, Cardiff was resistant to gentamicin but sensitive to tetracycline and trimethoprim.

Anti-Bacterial Agents↗

The colonization of solid PVC surfaces and the acquisition of resistance to germicides by water micro-organisms.

Six common water bacteria were examined for their ability to colonize polyvinyl chloride (PVC) surfaces, survive various germicidal treatment, and re-establish themselves in sterile distilled water (SDW). For each test, two 30.4 cm PVC pipes attached to a 90 degrees PVC elbow were filled with 600 ml of distilled water inoculated with either Pseudomonas aeruginosa, Ps. cepacia, Ps. mesophilica, Acinetobacter anitratus, Mycobacterium chelonae or M. chelonae var. abscessus. After 8 weeks contaminated water was removed and the pipes were exposed to 600 ml of 1:213 iodophor disinfectant (ID), 1:128 phenolic detergent (P), 1:256 quaternary ammonium compound (QA), stock iodophor antiseptic (IA), 2% formaldehyde (F), 10-15 ppm free chlorine (C), 2% glutaraldehyde (G) and 70% ethanol (E). These germicides were periodically sampled, neutralized and examined for surviving organisms. After exposure for 7 d the germicides were removed and each pipe was refilled with SDW. This was assayed at 7 d intervals to determine microbial re-establishment. Samples were removed during microbial conditioning and examined by scanning electron microscopy (SEM). Pseudomonads were isolated directly from ID, QA, C, P and F, and mycobacteria from QA, IA, ID, P, G, C and F. Pseudomonas aeruginosa and Ps. cepacia survived in PVC pipes after 7 d of exposure to P, ID and C; Ps. mesophilica, after C and ID; and both mycobacteria, after C. SEM examination of PVC remnants revealed bacterial attachment and formation of extracellular material with embedded cells. These studies show that common water bacteria can attach and colonize the interior surface of PVC pipes and develop significant resistance to the action of certain germicides.(ABSTRACT TRUNCATED AT 250 WORDS)

Acinetobacter↗

Pseudocontamination of blood components with Burkholderia cepacia during quality controls.

We report on a pseudooutbreak of Burkholderia cepacia because of the use of a contaminated disinfectant during quality controls in a university blood bank. No septic reactions associated with transfusions had been reported in patients over the last 6 months. Analysis of the individual quality control procedures showed that a disinfectant based on a quaternary ammonium compound (QAC) had been used in order to disinfect the rubber stopper of the blood culture bottle. B. cepacia was found in a sample taken from this disinfectant, which was prepared with concentrate and tap water according to the manufacturer's instructions. The four isolates (one in disinfectant and three in blood components) were found to be identical in their biochemical reactions and resistance patterns. QAC-based disinfectants are not efficacious against a part of the spectrum of gram-negatives and are therefore inadequate. After introduction of an alcohol-based preparation, no more cases of B. cepacia contamination have been identified.

Blood Component Transfusion↗

Glycopyrrolate compared with atropine in association with ketamine anaesthesia.

Atropine and glycopyrrolate given intravenously before the induction of a ketamine anaesthesia to diminish salivary secretion were compared for their effect on psychotomimetic side-effects, awakening time and heart rate. Though atropine is a tertiary amine that crosses the blood-brain barrier, which glycopyrrolate as a quaternary ammonium compound does not, it did not increase the incidence of psychotomimetic side-effects nor did it significantly prolong the awakening time after ketamine anaesthesia. During intubation the increase in heart rate was significantly higher following atropine than following glycopyrrolate.

Anesthesia↗

Selective inhibition of responses to carbachol and McN-A-343 in the rabbit vas deferens.

1. The effect of several selective muscarine receptor antagonists were evaluated on the responses of carbachol (CCh) and McN-A-343 (McN) during sympathetic nerve stimulation in the rabbit vas deferens. 2. The muscarine M1 receptor antagonist pirenzepine exhibited similar apparent pKB values for antagonism of the prejunctional inhibitory response of either CCh (pKB, 8.2) or McN (pKB, 8.5) on sympathetic nerve stimulation. 3. The muscarine M2 receptor antagonists, pancuronium and the bisalkyl ammonium compound 'C7/3-phth' were selective inhibitors of the postjunctional facilitatory response produced by CCh on sympathetic nerve stimulation. They were also 17- and three-fold, respectively, less potent against the inhibitory responses of McN on sympathetic nerve stimulation. The apparent pKB value of pancuronium on the inhibitory response produced by CCh did not differ significantly (P greater than 0.05) from that using McN. A similar finding was made for C7/3-phth. 4. Selective blockade of the inhibitory response to CCh with pirenzepine (0.03 or 0.5 mumol/L) did not significantly (P greater than 0.05) modify the apparent pKB value for pancuronium on the facilitatory response of CCh. 5. Selective blockade of the facilitatory response to CCh with a low concentration of pancuronium (0.5 mumol/L) did not significantly (P greater than 0.05) modify the apparent pKB value for pancuronium (30 mumol/L) on the inhibitory response of CCh. 6. It is suggested that CCh and McN activate the same prejunctional M1 muscarine receptor and that pancuronium is the most selective of the muscarine M2 receptor antagonists presently tested in this preparation for distinguishing between muscarine M1 and M2 receptors.

(4-(m-Chlorophenylcarbamoyloxy)-2-butynyl)trimethy↗

Disinfectant tolerance and antibiotic resistance in psychrotrophic gram-negative bacteria isolated from vegetables.

A total of 330 strains of psychrotrophic non-fermenting Gram-negative bacteria isolated from vegetables were studied. In spite of the wide range of antibiotic resistance occurring, less than 10% showed resistance patterns which included mezclocillin-ticarcillin-gentamicin or ceftizoxime-norfloxacin. Reductions of > 5 log10 in the numbers of cfu were found when these strains were exposed for 30 min to a quaternary ammonium compound (1% w/v).

Colony Count, Microbial↗

Effects of isoxazolium cations on some isolated muscle preparations.

1. The quaternary ammonium compound, N,t-butyl-5-methyl isoxazolium perchlorate (BIP), an anionic group reagent, initially causes contractions of the rat phrenic-nerve diaphragm, guinea-pig ileum and rabbit aortic strip preparations in vitro.2. In addition, the drug produces an irreversible block of indirectly elicited twitch responses in the diaphragm and of contractions induced by acetylcholine, methylfurmethide, dimethyl-phenylpiperazinium, 5-hydroxytryptamine (5-HT), histamine, angiotensin and pilocarpine in the ileum, while direct electrical stimulation of the diaphragm and contractions of the ileum to Ba and K ions are relatively unaffected.3. BIP is also an irreversible inhibitor of acetylcholinesterase but not of butyrylcholinesterase.4. On rabbit aortic strip preparations, responses to histamine, noradrenaline and 5-HT were differentially sensitive to irreversible blockade by BIP.5. Diphenhydramine, used in conditions which gave complete protection of the histamine response to irreversible block by dibenamine, did not protect against the blocking action of BIP but increased the blockade.6. These results suggest that BIP reacts covalently with anionic groups which mediate receptor initiated stimuli. The isoxazolium group may be useful in conferring irreversible properties by its substitution in drug molecules for the pyrrole or pyrrolidine group.

Acetylcholine↗

Pharmacological analysis of muscarinic receptors coupled to oxyntic cell secretion in the mouse stomach.

In the light of recent attempts to subclassify muscarinic receptors, agonist-antagonist interactions at muscarinic receptors have been re-examined using improved techniques, on the mouse, isolated, lumen-perfused stomach gastric acid assay. Using 5-methylfurmethide as the muscarinic agonist, the pKB estimated for atropine was significantly lower on the stomach assay (7.78) than on the guinea-pig trachea (8.93). However pKB values for N-methylatropine, the quaternary ammonium derivative of atropine, at concentrations producing dose-ratios above 20 on the stomach assay (pKB = 9.67), and over the full concentration range studied on the trachea (pKB = 9.69) were not significantly different. The deviation from simple competitive behaviour at low dose-ratios with N-methylatropine in the stomach assay is consistent with the effects of a saturable uptake mechanism for quaternary ammonium compounds. The pKB values for pirenzepine on the stomach (6.67) and the trachea (6.87) were not significantly different suggesting that pirenzepine behaves more like N-methylatropine in terms of expressed affinity. We conclude that the oxyntic cell muscarinic receptors are homogeneous with those in the guinea-pig trachea. An initial exploration suggests that there is a relationship between the lipophilicity (log P) of the antagonists and the degree of apparent underestimation of antagonist affinity in the stomach assay. This supports the hypothesis that the underestimation of antagonist affinity is due to the loss of antagonist into the gastric secretion from the receptor compartment. Apparently, relatively selective inhibition of acid secretion, compared to atropine, could be explained without the need to postulate heterogeneity of muscarinic receptor populations.

Animals↗

The refurbished pulse generator.

With the advent of long-life lithium pulse generators, normally functioning pulse generators with a potential life of more than five years have been removed from patients and become available for re-implantation. Although pulse generator refurbishing is widely employed, the practice has not been accepted in the United States. At The Royal Melbourne Hospital, all lithium pulse generators removed because of patient death or other causes were washed in a quaternary ammonium compound and soaked in formaldehyde. Pulse generators were than electronically tested and, if within specification, were made available for re-implantation. Pulse generators were then washed under sterile conditions in distilled water and gas-sterilized with ethylene oxide. Between 1975 and 1978 (48 months), 600 pulse generators were implanted and 93 pulse generators removed. There were 56 deaths, 22 cases of pre-erosion, erosion, or infection and 15 elective removals either due to lead problems or impending power source depletion. Eight-three (89%) pulse generators were refurbished (14% of total implants). This included 12 pulse generators refurbished on two occasions. Ten pulse generators were returned to manufacturers, seven because of impending power source depletion, two with suspected electronic faults, and one with a damaged case. Two complications occurred in patients with refurbished pulse generators. An infective process present with the previous pulse generator spread to a new pocket. The other pulse generator was removed 35 months post second implantation because of impending power source depletion. Primary infection or unusual tissue reactions did not occur. Pulse generator refurbishing as described was found to be a safe and economic procedure.

Electric Power Supplies↗