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The EcoRI restriction endonuclease, covalently closed DNA and ethidium bromide.

The reactions of the EcoRI restriction endonuclease on the covalently closed DNA of plasmid pMB9 were studied in the presence of ethidium bromide. At the concentrations of ethidium bromide tested, which covered the range over which the DNA is changed from negatively to positively supercoiled, the dye caused no alteration to the rate at which this enzyme cleaved the covalently closed DNA to yield the open-circle form, but the rate at which these open circles were cleaved to the linear product could be inhibited. The fluorescence change, caused by ethidium bromide binding with different stoichiometries to covalently closed and open-circle DNA, provided a direct and sensitive signal for monitoring the cleavage of DNA by this enzyme. This method was used for a steady-state kinetic analysis of the reaction catalysed by the EcoRI restriction enzyme. Reaction mechanisms where a complex between DNA and Mg2+ is the substrate for this enzyme were eliminated, and instead DNA and Mg2+ must bind to the enzyme in separate stages. The requisite controls for this fluorimetric assay in both steady-state and transient kinetics studies, and its application to other enzymes that alter the structure of covalently closed DNA, are described.

DNA↗

Neuromuscular recovery following rocuronium bromide single dose in infants.

BACKGROUND: Rocuronium bromide, a steroid nondepolarizing muscle relaxant, has a rapid onset and an intermediate duration of action in infants, children and adults. However, clinical evidence shows a longer duration of recovery in small infants. The aim of this study was to investigate the influence of age on rocuronium recovery during the first year of life. METHODS: ASA I-II infants, scheduled for elective surgery under general anaesthesia and intubation were included after ethics committee approval and parents' written consent. According to age the patients were randomly allocated to receive either 0.45 mg.kg(-1) or 0.6 mg.kg(-1) rocuronium bromide in three age-groups: (A) 0-1 month, (B) 2-4 months and (C) 5-12 months. After induction with thiopentone (5-7 mg.kg(-1)), anesthesia was maintained with isoflurane without opioids. Prior to surgery, caudal block with bupivacaine (0.125%) 1.0 ml.kg(-1) and paracetamol 25 mg.kg(-1) rectally were given for analgesia. Efficacy variables were intubation conditions 60 s after administration of muscle relaxant (T(0)) and recovery of neuromuscular blockade measured as T(1) at 10, 25, 50 and 75 % of baseline, train-of-four (TOF) of 0.7 and Recovery Index (RI). Data were characterized by summary statistics and analysis of variance. RESULTS: A total of 61 infants with a median age range of 67 (2-364) days were included. Intubation conditions were excellent or good in all dose and age groups. T(0) in group A was reached in a range of 15-30 s, in others up to 60 s. T(1) recovery (T75) after 0.45 mg.kg(-1) was 56.4 +/- 16 (A), 62.7 +/- 32 (B) and 45.8 +/- 18 (C) min. Recovery times for of 0.6 mg.kg(-1) were 100.8 +/- 35 (A), 70.6 +/- 19 (B) and 63.4 +/- 21 (C) min, respectively. The TOF ratio (0.7) was 62.3 +/- 18 (A), 64.1 +/- 27 (B) and 43.7 +/- 12 (C) min using 0.45 mg.kg(-1) compared with 94.8 +/- 31 (A), 63.8 +/- 14 (B) and 67.5 +/- 18 (C) min with 0.6 mg.kg(-1). The differences of T75 and TOF 0.7 in A and C were significant (P </= 0.05). CONCLUSIONS: Recovery of muscle relaxation using rocuronium bromide under isoflurane anesthesia in infants differs widely and shows great differences between age groups as well as dose regimen. A dose of 0.6 mg.kg(-1) resulted in a significantly longer duration of action in group A. The reduced dose of 0.45 mg.kg(-1) resulted in rapid and good relaxation in all infants without very long lasting effects. Reduced doses of rocuronium should be used in newborns and small infants.

Age Factors↗

The effects of topical ocular application of 0.25% demecarium bromide on serum acetylcholinesterase levels in normal dogs.

OBJECTIVE: To assess the effects of topical ocular application of 0.25% demecarium bromide on serum acetylcholinesterase (AChE) levels in normal dogs. ANIMALS: Nine adult mixed breed dogs weighing between 18 and 27 kg. PROCEDURES: Fifty micro L of 0.25% demecarium bromide were applied to one eye of each dog every 8 h for 6 days. Blood was analyzed for AChE levels prior to commencement of eye drops, and at 45 min, 1 h 45 min, 4 h 45 min, 1 day, 3 days, and 7 days following commencement of eye drops using a 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB) reaction. RESULTS: Acetylcholinesterase levels declined over the first 24 h following commencement of demecarium administration in most dogs. This decline was highly variable and was statistically significant by 24 h. In some individuals AChE levels were suppressed to levels approaching clinical toxicity. By day 3 AChE levels had risen to levels above baseline in most dogs. CONCLUSIONS: Topical ocular application of demecarium causes transient suppression of systemic acetylcholinesterase levels in most dogs. Acetylcholinesterase levels generally do not fall to toxic levels, but may do so in certain individuals. Demecarium bromide eye drops generally do not cause AChE toxicity, but dogs receiving such therapy should be monitored for signs of AChE toxicity, and concomitant use of other AChE inhibitors should be avoided.

Acetylcholinesterase↗

[Comparison of emepronium bromide with intravesical administration of lidocaine gel in women with urge incontinence].

In the treatment of urge incontinence, drug therapy predominates. In a prospective randomised study on treatment of females with motor urge incontinence, intravesically administered lidocaine-gel and oral emepronium bromide were compared using clinical and urodynamic parameters. From 30 patients, 15 women were treated for 3 weeks with repeated intravesical lidocaine gel instillations whilst the other 15 had a peroral emepronium bromide medication. The intravesical lidocaine gel installation proved more efficient compared with peroral emepronium bromide therapy with regard to the parameters of imperative micturition, occurrence of detrusor contractions as well as number of urinary frequencies per diem. Intravesical application of lidocaine-gel is particularly indicated in patients with increased sensitivity against parasympathicolytics and anticholinergics.

Administration, Intravesical↗

The dynamics of Br(2Pj) formation in the photodissociation of vinyl and perfluorovinyl bromides.

The photodissociation dynamics of vinyl bromide and perfluorovinyl bromide have been investigated at 234 nm using a photofragment ion imaging technique coupled with a state-selective [2+1] resonance-enhanced multiphoton ionization scheme. The nascent Br atoms stem from the primary C-Br bond dissociation leading to the formation of C2H3(X) and Br(2Pj;j=1/2,3/2). The obtained translational energy distributions have been well fitted by a single Boltzmann and three Gaussian functions. Boltzmann component has not been observed in the perfluorovinyl bromide. The repulsive 3A'(n,sigma *) state has been considered as the origin of the highest Gaussian components. Middle translational energy components with Gaussian shapes are produced from the 1A"(pi,sigma*) and/or 3A"(pi,sigma*) which are very close in energy. Low-energy Gaussian components are produced via predissociation from the 3A'(pi,pi*) state. The assignments have also been supported by the recoil anisotropy corresponding to the individual components. It is suggested that intersystem crossing from the triplet states to the ground state has been attributed to the Boltzmann component and the fluorination reduces the probability of this electronic relaxation process.

Journal Article↗

Interaction of ethidium bromide with heparin.

The reactions of hydrated electrons produced during pulse radiolysis have been utilized to investigate the binding of ethidium bromide to heparin. Complexes of ethidium bromide and heparin can be dissociated with salt. Divalent cations are more effective than monovalent cations in this respect. Pulse-radiolysis investigations at different temperatures indicate that the thermodynamic parameters governing the interaction of ethidium bromide with heparin are deltaH' = 11-6 kcal mole-1 and deltaS' = 42-6 cal deg-1 mole-1.

Electrons↗

Evaluation of weather conditions for methyl bromide ambient air monitoring in Monterey, Santa Cruz, and Kern Counties in the year 2000.

Weather condition is one of the most important factors affecting spatial and temporal distributions of air pollutants, especially for short-term air dispersion. Abnormal weather conditions might lead to higher or lower ambient air concentrations than they would be under normal weather conditions. Therefore, testing for normality of weather conditions during the air monitoring period is an essential step for evaluating ambient air monitoring results. In this paper, a distance method was used to select a most representative weather station from the available candidates. An array of meteorological elements were identified that affect air dispersion and transportation. A statistical method was used to determine whether the weather conditions during the air monitoring period were significantly different from that of previous years. Using methyl bromide ambient air monitoring as a case study, this paper documents the methods, procedures, and results of weather analysis for Monterey, Santa Cruz, and Kern Counties during ambient air monitoring periods for methyl bromide in the year 2000. With a few exceptions, the meteorological elements and atmospheric stability factors, such as wind speeds, wind directions, and stability classes, during the monitoring period were in the normal range. Although there were higher frequencies of stable atmospheric conditions in Monterey/Santa Cruz Counties than in Kern County, weather conditions during the monitoring period were not significantly different from normal local weather conditions of previous years. Consequently, the subchronic air concentrations observed during the ambient air monitoring periods for methyl bromide in the year 2000 was taken under typical weather conditions of those areas at that time of the year.

Air Pollutants↗

Manufacture, characterization, and pharmacodynamic evaluation of engineered ipratropium bromide particles.

PURPOSE: The intent of this research was to generate and characterize respirable particles of ipratropium bromide (IPB), a short-acting anticholinergic bronchodilator, to achieve demonstrable sustained-release properties. The value of a long-acting anticholinergic agent is evident in the use of tiotropium for the treatment of chronic obstructive pulmonary disease. METHODS: Hollow, spherical particles of ipratropium bromide suitable for inhalation were generated using a spray-drying process and characterized by laser diffraction particle size analysis, scanning electron microscopy, dynamic vapor sorption, X-ray diffraction, differential scanning calorimetry, thermogravimetric analysis, and dissolution testing. Experimental design techniques were used to identify critical process parameters and optimize the spray drying process. Pharmacodynamic studies were conducted to determine duration of effect. RESULTS: Crystalline, stable, respirable particles with a range of dissolution profiles were manufactured by application of polylactic acid (PLA) coatings of 1, 5, 10, 15, 30, and 50% w/w. A novel, robust, modified Type IV dissolution method discriminated between formulations and guided their development. Preliminary studies in guinea pigs indicated an increased duration of bronchodilatory effect for 30% PLA-coated particles (56.3 min) particles compared with IPB powders alone (11.0 min). CONCLUSIONS: Sustained-release respirable particles of ipratropium bromide were developed using a PLA spray coating approach and a trend for increased duration of effect was demonstrated in guinea pigs.

1,2-Dipalmitoylphosphatidylcholine↗

The efficacy of nebulized albuterol/ipratropium bromide versus albuterol alone in the prehospital treatment of suspected reactive airways disease.

OBJECTIVE: Ipratropium bromide has demonstrated efficacy when added to albuterol for the treatment of reactive airways disease (RAD). Its prehospital use has not been explored. METHODS: A before-and-after design was used. Prehospital and emergency department (ED) medical records were examined retrospectively six months before and six months after institution of a new protocol, which allowed the addition of ipratropium bromide to all nebulized treatments with albuterol. Primary outcome measures included: changes in vital signs (heart rate, respiratory rate, oxygen saturation), clinical improvement as assessed by paramedics, and admission rates. RESULTS: A total of 371 patients were included (n = 192 albuterol alone, n = 179 ipratropium/albuterol). There was no statistically significant difference between groups with regard to the change in heart rate, respiratory rate, or oxygen saturation. In addition, there was no difference in the proportion of patients with clinical improvement or deterioration as assessed by paramedics. There was no statistically significant difference in the admission rate from the ED except in the subgroup of patients using a metered-dose inhaler at the time of illness. Of note, more than one-third (133/371) of the patients were ultimately determined to have a diagnosis other than RAD, the majority of whom were diagnosed as having cardiac disease. CONCLUSIONS: The addition of ipratropium bromide to albuterol for the prehospital treatment of RAD does not appear to result in clinical outcome improvements. A substantial number of patients enrolled in the study were diagnosed as having cardiac disease.

Administration, Inhalation↗

Determination of acute toxic effects in mice following exposure to methyl bromide.

An inhalation system was designed and constructed for acute nose exposure of mice to methyl bromide. Animals were exposed for 1 h to concentrations ranging from 0.87 to 5.93 mg/l. Mice exposed to concentrations up to 1.72 mg/l did not exhibit any indication of developing a toxic response. Animals exposed to concentrations to 2.20 and 2.70 mg/l exhibited significantly decreased lung and liver weights when compared to controls. Animals exposed to concentrations above 3.50 mg/l exhibited kidney lesions. At concentrations of 3.82 mg/l and above, animals exhibited abnormal clinical signs, weight loss, and mortality. In addition, at 4.70 mg/l, a liver lesion was observed. At concentrations above 5.77 mg/l, pathological changes were observed in the color and a decreased motor coordination was evident. Methyl bromide exposures of up to 3.82 mg/l did not affect the ability of mice to recall a single-task passive-avoidance test. The 1-h LC50 of methyl bromide in mice via inhalation was determined to be 4.68 mg/l (approximately 1200 ppm). The dose-response curve was quite steep and the LC10 to LC90 range of mortality was contained within a doubling of concentration.

Animals↗

Dose-morality and large-scale studies for controlling codling moth (Lepidoptera: Tortricidae) eggs on 'd'Agen' plums by using methyl bromide.

Codling moth, Cydia pomonella (L.), eggs on 'd'Agen' cultivar of plums, Prunus salicina Lindl., were treated with methyl bromide to determine if a quarantine treatment could be developed so that the plums could be exported to Japan from the United States. Small-scale tests consisted of treating codling moth eggs on plums with various doses of methyl bromide at 20 degrees C for 2 h. Small-scale tests showed that 0- to 24-h-old eggs of codling moth on the plums were controlled by doses > 22.5 g/m3. Because testing showed that 48 g/m3 had no adverse effect on the quality of plums, this dose was chosen for large-scale testing to establish the quarantine dose. Large-scale tests consisted of treating plums at 18.5 degrees C for 2 h using methyl bromide at a dose of 48 mg/liter. Large-scale tests showed that the dose of 48 g/m3 killed all 0- to 24-h-old codling moth eggs exposed on plums in packing cartons without affecting the quality of the plums.

Animals↗

Effect of ethidium bromide on transplanted virus-induced tumor cells.

Ethidium bromide (2,3-diamino-5-ethyl-6-phenylphenanthridinium bromide) significantly inhibited the RNA-dependent DNA polymerase of types A and C particles isolated from transplanted adenovirus 12-induced tumors of CBA mice. It was also cytotoxic for an established in vitro line of adenovirus 12-induced tumor cells of CBA mice and caused cell death, inhibition of [3H]thymidine uptake, and a significant reduction of cells in metaphase. Ethidium bromide significantly inhibited the in vivo growth of transplanted adenovirus 12-induced tumor cells of CBA mice, simian virus 40-induced tumor cells of hamsters, and murine leukemia virus-induced lymphoma cells of BALB/c mice. The compound may have exerted the antitumor activity by selectively affecting oncornavirus in the tumor cells.

Adenoviridae↗

A study of the interaction between ethidium bromide and rye chromatin: comparison with calf thymus chromatin.

We studied the interaction of ethidium bromide with rye and calf thymus chromatin. Both types of chromatin have the same dye accessibility, which is about 50% of that of DNA. From this result we conclude that the molecular structure of these two chromatins is similar. For rye, the extraction of H1 produces no change in the binding of ethidium bromide. The subsequent extraction of H2A and H2B produces a 14% increase in the binding, and the removal of H3 and H4, another 54% increase. At this stage, the number of binding sites is still less than that of DNA. This is presumably due to the presence of some tightly bound non-histones. Thus, the arginine-rich histones and the tightly bound non-histones are most responsible for limiting the binding of ethidium bromide to rye chromatin.

Animals↗

Heat-induced DNA relaxation in vitro by mouse DNA topoisomerase I in the presence of ethidium bromide.

In general, covalently-closed circular DNAs that are relaxed by DNA topoisomerase I at higher temperatures are more negatively (or less positively) supercoiled when analyzed at the same temperature. We found that in the presence of ethidium bromide the reaction products were less negatively supercoiled at higher temperatures. Consequently, the linking number of substrate DNA changed reversibly with temperature-shift: DNA relaxation occurred on temperature shift-up, and re-supercoiling on temperature shift-down. Analyses of the migration of covalently closed circular DNA on agarose gel electrophoresis and of the fluorescence intensity of ethidium bromide bound to DNA indicated decrease in binding of the drug to DNA at higher temperatures. Thus, the thermal effects on the topology of the reaction products of mouse DNA topoisomerase I in the presence of ethidium bromide can be explained by the temperature-sensitive interaction of the drug with DNA.

Animals↗

The effects of pipecuronium bromide on intracranial pressure and cerebral perfusion pressure.

Twenty patients with expansive pathologic intracranial lesions, who were anesthetized with thiopental, nitrous oxide in oxygen, and fentanyl and mechanically ventilated to ensure normocarbia, received pipecuronium bromide 70 microg/kg i.v. Intracranial pressure (ICP), heart rate, arterial pressure, central venous pressure (CVP), EKG, and end-tidal CO2 were simultaneously recorded for 5 min before and for 15 min after administration of the muscle relaxant. No statistically significant changes in ICP and cerebral perfusion pressure were observed after administration of pipecuronium bromide. Cardiovascular stability was maintained during the study period except for a small, although significant, decrease of the CVP from 5.7 +/- 2.5 (SEM) to 5.0 +/- 2.5 mm Hg. These results, together with the long-lasting muscular effect of pipecuronium bromide, suggest that this new neuromuscular blocking agent may be used for muscle relaxation during neurosurgical operations in patients who have normal intracranial pressure at the time of administration of the drug.

Journal Article↗

Leisingera methylohalidivorans gen. nov., sp. nov., a marine methylotroph that grows on methyl bromide.

A marine methylotroph, designated strain MB2T, was isolated for its ability to grow on methyl bromide as a sole carbon and energy source. Methyl chloride and methyl iodide also supported growth, as did methionine and glycine betaine. A limited amount of growth was observed with dimethyl sulfide. Growth was also noted with unidentified components of the complex media marine broth 2216, yeast extract and Casamino acids. No growth was observed on methylated amines, methanol, formate, acetate, glucose or a variety of other substrates. Growth on methyl bromide and methyl iodide resulted in their oxidation to CO2 with stoichiometric release of bromide and iodide, respectively. Strain MB2T exhibited growth optima at NaCl and Mg2+ concentrations similar to that of seawater. Phylogenetic analysis of the 16S rDNA sequence placed this strain in the alpha-Proteobacteria in proximity to the genera Ruegeria and Roseobacter. It is proposed that strain MB2T (= ATCC BAA-92T = DSM 14336T) be designated Leisingera methylohalidivorans gen. nov., sp. nov..

Alphaproteobacteria↗

Neuromuscular blockade of fazadinium bromide (AH 8165) in renal failure patients.

Fazadinium bromide (AH 8165) is a new non-depolarizing neuromuscular blocking agent with rapid onset of action and intermediate duration of effect. The characteristics of the neuromuscular blockade induced were compared in normal and renal failure patients by monitoring the mechanical responses of thumb adductor muscle to ulnar nerve stimulation. The data suggest that renal failure, even in anephric patients, does not influence the depth of neuromuscular relaxation when judged from the identical dose-effect curves in both groups of patients. Moreover, the duration of the neuromuscular block was not significantly increased in end-stage kidney failure patients, whether the initial or late phases of the spontaneous recovery of neuromuscular transmission were considered, and this occurred over the entire therapeutic range. However, additional incremental injections of fazadinium bromide should be used cautiously, as evidence appeared of prolonged duration of action. This agent, fazadinium bromide, may therefore present useful advantages to other non-depolarizing neuromuscular blocking agents as to clinical safety and effects in patients with renal insufficiency.

Adolescent↗

Inhibition of methoxamine-induced bronchoconstriction by ipratropium bromide and disodium cromoglycate in asthmatic subjects.

We compared the effects of pretreatment with saline, ipratropium bromide, and disodium cromoglycate (DSCG) on bronchoconstriction induced by methoxamine--an alpha-adrenoceptor agonist, in asthmatic subjects. All 12 patients bronchoconstricted in response to methoxamine after saline. The PD20 (the dose of methoxamine causing a 20% fall in forced expiratory volume in 1 s [FEV1]) ranged from 0.3-18 mumol. Ipratropium bromide (200 micrograms administered by aerosol) significantly inhibited (P less than 0.05) the response to methoxamine in all patients without producing significant changes in the mean baseline lung function. The mean PD20 for methoxamine after saline was 6.8 mumol and 95% confidence limits (CL) were 3.6, 12.7 mumol. The mean PD20 for methoxamine after ipratropium bromide was 35.4 (95% CL 28.8, 43.6) mumol. DSCG also produced significant (P less than 0.05) shifts to the right in the methoxamine dose response curves, but did not affect resting airway calibre as measured by the FEV1. The mean PD20 for methoxamine increased from 3.3 mumol (95% CL 1.1, 10.0 mumol) after saline to 25.1 mumol (95% CL 14.1, 44.6) after DSCG pretreatment. These findings suggest that alpha-adrenoceptors in the airways of asthmatic subjects may be located at sites other than smooth muscle--possibly on mast cells but more likely on nerve endings and/or parasympathetic ganglia.

Adult↗