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Analysis of plasma catecholamines by high-performance liquid chromatography with fluorescence detection: simple sample preparation for pre-column fluorescence derivatization.

Analysis of plasma catecholamines (norepinephrine, epinephrine and dopamine) by high-performance liquid chromatography using 1,2-diphenylethylenediamine as a fluorescent reagent is described. We have developed an automatic catecholamine analyser, based on pre-column fluorescence derivatization and column switching. The analysis time for one assay was 15 min. The correlation coefficients of the linear regression equations were greater than 0.9996 in the range 10-10,000 pg/ml. The detection limit, at a signal-to-noise ratio of 3, was 2 pg/ml for dopamine. A new method of sample preparation for the pre-column fluorescence derivatization of plasma catecholamines was used. In order to protect the catecholamines from decomposition, an ion-pair complex between boric acid and the diol group in the catecholamine was formed at a weakly alkaline pH. The stabilities of plasma catecholamines were evaluated at several temperatures. After complex formation, the catecholamines were very stable at 17 degrees C for 8 h, and the coefficients of variation for norepinephrine, epinephrine and dopamine were 1.2, 4.2 and 9.3%, respectively.

Autoanalysis↗

Sensitive determination of carbohydrates labelled with p-nitroaniline by capillary electrophoresis with photometric detection using a 406 nm light-emitting diode.

p-Nitroaniline was explored as a derivatising reagent for UV absorbance detection of carbohydrates after separation by CE. This derivatising agent has three advantages: first, it has excellent water solubility; second, it has high molar absorptivity; and third, it is possible to obtain sensitive detection using a UV or blue light-emitting diode (LED) as the light source. The labelling reaction took less than 30 min to complete with high reaction yield. The separation process was modelled and optimised using an artificial neural network. Nine carbohydrates were separated by a CE system within 16 min using a 0.17 M boric acid buffer at pH 9.7. On-column LED detection at 406 nm allowed the detection of carbohydrates with good detection limits (<1.1 microM or 8.8 fmol) and reproducible quantification in the concentration range of 2.6-200 microM. This method was applied successfully to the determination of component carbohydrates in some food samples.

Aniline Compounds↗

Germanium does not substitute for boron in cross-linking of rhamnogalacturonan II in pumpkin cell walls.

Boron (B)-deficient pumpkin (Cucurbita moschata Duchesne) plants exhibit reduced growth, and their tissues are brittle. The leaf cell walls of these plants contain less than one-half the amount of borate cross-linked rhamnogalacturonan II (RG-II) dimer than normal plants. Supplying germanium (Ge), which has been reported to substitute for B, to B-deficient plants does not restore growth or reduce tissue brittleness. Nevertheless, the leaf cell walls of the Ge-treated plants accumulated considerable amounts of Ge. Dimeric RG-II (dRG-II) accounted for between 20% and 35% of the total RG-II in the cell walls of the second to fourth leaves from Ge-treated plants, but only 2% to 7% of the RG-II was cross-linked by germanate (dRG-II-Ge). The ability of RG-II to form a dimer is not reduced by Ge treatment because approximately 95% of the monomeric RG-II generated from the walls of Ge-treated plants is converted to dRG-II-Ge in vitro in the presence of germanium oxide and lead acetate. However, dRG-II-Ge is unstable and is converted to monomeric RG-II when the Ge is removed. Therefore, the content of dRG-II-Ge and dRG-II-B described above may not reflect the actual ratio of these in muro. (10)B-Enriched boric acid and Ge are incorporated into the cell wall within 10 min after their foliar application to B-deficient plants. Foliar application of (10)B but not Ge results in an increase in the proportion of dRG-II in the leaf cell wall. Taken together, our results suggest that Ge does not restore the growth of B-deficient plants.

Boric Acids↗

Carbamylation of alkaline mesentericopeptidas.

The effects of carbamylation with potassium cyanate, and methylation with methyl p-nitrobenzene sulphonate on the mesentericopeptidase activity are studies. The treatment with potassium cyanate causes the enzyme to lose its activity towards ester substrates and casein. The specific reagent N-trans-cinnamoylimidazole does not acylate the active site in the carbamylated enzyme. The pH dependence of the rate of inactivation indicates that an ionizing group of pK = 7.3, probably the protonated imidazole group of the active site histidine, is involved in the reaction. The competitive inhibitor boric acid protects mesentericopeptidase against inactivation with potassium cyanate. These suggest that the active site residues are modified in the unprotected enzyme. Sixty per cent of the enzyme activity toward N-acetyl-L-tyrosine ethyl ester was restored after treatment of the carbamylated mesentericopeptidase with 1 M hydroxylamine hydrochloride. Circular dichroism spectra show that the carbamylation does not change markedly the native protein conformation.

Acylation↗

The granulation of ternary mixtures: the effect of the wettability of the powders.

Granules have been prepared by a standard method from ternary mixtures of lactose, boric acid and either sulphanilamide, heavy kaolin or salicylic acid, using an aqueous solution of polyvinylpyrrolidone as binder. The granules were examined for size distribution and for the work required to crush larger granules from each batch. For the same binder volume the mean granule sizes and resistance to crushing of the granules increased as the wettability of the third component increased. The detailed results are discussed in relation to powder wetting and the packing properties of powder mixtures.

Boric Acids↗

The granulation of ternary mixtures: the effect of the solubility of the excipients.

The importance of excipient solubility in wet granulation has been shown using mixtures of lactose and boric acid containing third components of differing solubility characteristics. Sucrose dissolved in the binder solution and increased the amount of liquid available to form liquid bridges between other solid particles and therefore increased the number of solid bridges when the damp mass was dried. Thus, stronger, larger granules were formed. The results obtained with Sta-Rx 1500 and magnesium carbonate powders were also dependent upon the extent of solubility of these powders in the granulating solution.

Boric Acids↗

Motility of three strains of Proteus mirabilis (G9, P11 and PM5C) in liquid suspension under different environmental conditions.

The effect of compounds on the motility of Proteus mirabilis swarmer cells varies from one strain to another. The effect of compounds on the motility of swarmer cells is mainly at higher concentrations than the concentration used to inhibit swarming. Boric acid only affects the motility of strain G9 swarmer cells, whereas sodium deoxycholate prevented the motility of swarmer cells for three strains. Some antibiotics show their effect on the motility of swarmer cells in anaerobic areas, by slowing the movement of swarmer cells, followed by stopping the movement after a period of time or disappearance of the cells. The differentiation between the strains of Proteus species seems to be better in liquid suspension than on the solid medium.

Anti-Bacterial Agents↗

Separation of ibuprofen, codeine phosphate, their degradation products and impurities by capillary electrophoresis. I. Method development and optimization with fractional factorial design.

A capillary electrophoresis method has been developed and optimized for the separation of ibuprofen, codeine phosphate and their main degradation products and impurities. In the course of developing the method, it was found that micellar electrokinetic capillary chromatography was necessary for the separation of the eleven peaks. A fractional factorial design was used for the optimization of the experiments. Six process parameters were varied at two levels: the concentration of sodium dodecyl sulfate (SDS), the pH, the concentration of acetonitrile, the concentration of boric acid, the field strength and the temperature. All these factors had a significant effect on the migration time and resolution. The optimum conditions were found to be a borate buffer of 40 mM H3BO3 at pH 10 with the addition of 40 mM SDS and 9% acetonitrile, a field strength of 515 V/cm and a temperature of 25 degrees C. This resulted in baseline separation of the eleven peaks within 12 min.

Acetonitriles↗

Otomycosis in Turkey: predisposing factors, aetiology and therapy.

Otomycosis usually requires long-term treatment and tends to recur. This study was performed on 87 patients with the clinical diagnosis of otomycosis and 20 controls in order to determine the pathogenic agents, predisposing factors and a cost-effective treatment. The predisposing factors included wearing head clothes (74.7 per cent), presence of dermatomycoses (34.5 per cent) and swimming (27.6 per cent). The most common pathogenic fungus was Aspergillus niger (44.8 per cent) in the otomycosis group. The only isolate was Candida albicans in the control group (2.5 per cent). We concluded that administration of four per cent boric acid solution in alcohol and frequent suction cleaning of the ear canal might be a cost-effective treatment for otomycosis since 77 per cent of the patients were treated effectively this way. Eighty per cent of the resistant cases had mixed fungal-bacterial infections, and 50 per cent of them had dermatomycoses. These resistant cases were treated by administration of tioconazole ointment.

Adolescent↗

[Comparative characteristics of bactericidal activity of various traditional and currently available antiseptics used in surgery].

Comparative study of the microbicidal activity of some traditional (hydrogen peroxide, boric acid, nitrofurazone) and currently available (Baliz-2, dioxidine, chlorhexidine bigluconate, pervomur) antiseptics against the most commonly encountered causative agents of purulent surgical infection showed 0.8%, 1.2%, and 2.4% pervomur solutions to possess the highest bactericidal and sporicidal activity. This circumstance allowed weak (0.8 and 1.2%) solutions of pervomur to be used in the clinic in 68 patients for the management of purulent postoperative complications and localized purulent infection, which improved the results of treatment significantly.

Anti-Infective Agents, Local↗

Current therapy of vulvovaginitis.

Trichomoniasis is reliably treated with a single 2-g dose of metronidazole; however, with this regimen simultaneous treatment of sexual partners is particularly important. Trichomoniasis in pregnant women, who should not receive metronidazole, might be treated initially with clotrimazole vaginal suppositories, which appear to cure about 50% of cases. Topical antifungal agents of the imidazole class are superior to polyenes in treating vulvovaginal candidiasis. Boric acid powder applied intravaginally in gelatin capsules for 14 days appears as effective as imidazoles. Nonspecific vaginitis is now recognized as involving infection with anaerobic bacteria of the vaginal flora as well as Gardnerella vaginalis. The condition is most successfully treated with a seven-day course of metronidazole, which probably acts by eradicating the anaerobes. In addition, metabolites of metronidazole may act directly on G. vaginalis. Sulfanilamide-aminacrine-allantoin preparations are much less effective than specific therapies and have no role in the treatment of vulvovaginitis.

Antifungal Agents↗

Activated sludge immobilization using the PVA-alginate-borate method.

The PVA-alginate-borate method was used successfully to immobilize activated sludge. The optimum polyvinyl alcohol (PVA) concentration in the immobilized sludge was determined to be 10-12.5%. A minimal alginate of 1% in the beads was needed to prevent bead agglomeration. If the pH of the saturated boric acid was adjusted to 7.0 prior to allowing droplets of the mixture of PVA, alginate and sludge to enter the solution, a high level of sludge activity could be maintained in the beads formed. During the continuous operation of a fluidized bed reactor, with hydraulic retention time (HRT) in the range of 24-3 h (BOD loading from 0.176 to 0.766 kg/d/m3) and an aeration rate at 1.0 L/min, more than 90% of BOD5, NH4(+)-N and TKN were removed. The immobilized sludge exhibited satisfactory mechanical stability without apparent breakage.

Alginates↗

A new simple method for biotyping Candida albicans.

A simple method which allows rapid, reproducible biotyping of Candida albicans isolates, and is suitable for use in any diagnostic medical microbiology laboratory is described. This system comprises three tests, the API ZYM system, the API 20C system, and a plate test for resistance to boric acid. The system differentiated a possible 234 biotypes, of which 33 were found amongst the 130 isolates of C. albicans taken from oral, genital and skin sites. Major biotypes found amongst oral and non-oral isolates were similar, although differences were found amongst minor biotypes. The new system was found to be reproducible, discriminatory, reasonably fast (48 h), easy to perform, and had no requirement for specialised equipment. Hence this biotyping system is suitable for use in diagnostic microbiology laboratories for epidemiological investigations of C. albicans infections.

Boric Acids↗

Agarose electrophoresis of DNA in discontinuous buffers, using a horizontal slab apparatus and a buffer system with improved properties.

Using a horizontal slab apparatus with a buffer in the reservoirs at the level of the gel ("sea-level electrophoresis"), the retrograde discontinuous buffer system reported by Wiltfang et al. for sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of proteins was applied to DNA electrophoresis. This application yielded the advantages of an increased displacement rate of the moving boundary front and a decrease in the concentration of the counterion base in the resolving phase, which yielded reduced relative mobility values at equivalent gel concentrations and practicable low buffer concentrations. The change of relative mobilities (Rf) with a variation of field strength is decreased compared to that of the migration rate in the continuous Tris-boric-acid-EDTA (TBE) buffer and thus the robustness of the system is improved, as well as the efficiency of separation. The system of Wiltfang et al. has in common with previously described discontinuous DNA system, that it is able to stack DNA from dilute samples and is insensitive to sample components with lower net mobilities than DNA, such as acetate. However, the variance of Rf at constant current density in the discontinuous buffer system is not improved over that of the migration rate at constant field strength in the continuous TBE buffer.

Boric Acids↗

Isotope ratios of molybdenum determined by thermal ionization mass spectrometry for stable isotope studies of molybdenum metabolism in humans.

Methods were developed to separate and purify Mo from biological samples and to measure isotopic ratios in 1 microgram of Mo. A magnetic sector, thermal ionization mass spectrometer was used with simultaneous collection of five isotopes. Isotopic ratios were corrected for mass fractionation by iterative normalization using the 96/98 ratio. Ion beam intensity was enhanced by using a double-filament configuration, loading samples onto evaporation filaments with silica gel and boric acid. A triple-isotope-dilution approach was used, so the method could be applied to two-tracer studies of Mo metabolism in human subjects. 94Mo was added to samples prior to purification to quantify the total Mo content of samples and to determine the amounts of enriched 97Mo and 100Mo appearing in urine and fecal samples of study participants. The three ratios, 94/98, 97/98, and 100/98, were determined with within-run precision of from 0.06 to 0.10% (RSD). Precision of the ratios between replicates was from 0.05 to 0.08%.

Boric Acids↗

Association of reversible alopecia with occupational topical exposure to common borax-containing solutions.

Boron is widely used in industrial materials, most frequently as the salt borax. Systemic exposure (eg, ingestion) to boron in boric acid been associated with reversible toxic alopecia among other manifestations. There is scant clinical literature on alopecia caused by topical exposure to boron. We observed a series of 3 patients in 2 workplaces who suffered reversible alopecia from cutaneous boron exposure. The scalp alopecia was global in 1 patient and patchy in 2 patients. Alopecia was completely reversed by elimination or reduction of exposure to boron-containing materials in all 3 patients. We conclude that occupational topical exposure to boron in solutions may cause reversible alopecia.

Adult↗

Candida albicans strain differentiation in complete denture wearers.

Strain differentiation of 66 clinical isolates of Candida albicans obtained from healthy dentate and complete denture wearers was performed. Resistogram method based on differences in the resistance of C. albicans isolates to sodium selenite, boric acid, cetrimide, sodium periodate and silver nitrate was used for strain differentiation. Of the 32 potential strains that can be distinguished, 14 different resistogram strains of C. albicans were found among the 66 isolates tested. Strain-C--was the most predominant (24.3% of total isolates), while strain A-CDE was the least predominant (1.5%). The results showed no particular association of certain strains with Candida infections in complete denture wearers. Sensitivity to antifungal agents showed that isolates from different strains were most sensitive to amphotericin B and nystatin and least sensitive to miconazole.

Amphotericin B↗

Assessment of a short-term reproductive and developmental toxicity screen.

Short-term tests for reproductive and developmental toxicity are needed to provide preliminary data on the toxicity of chemicals about which little or no data exist. An ideal design would test all aspects of reproduction and identify the target process in a short time period. One potential design has been evaluated using four chemicals of varying reproductive/developmental toxicity. Swiss mice were mated for 3 days prior to chemical exposure to produce time-mated females for gestational exposure and to ascertain fertility of the untreated males. The group of time-mated females was treated during Gestation Days 8-14 and allowed to litter for observations through Postnatal Day (PND) 4. Endpoints observed included pup number and body weights on PND 0, 1, and 4 and number of uterine implantation sites on PND 4. A second group of females was dosed daily for 19 days. After 7 days, these females (n = 10/group) were cohabited with male mice who had been treated for 5 days prior to this second mating. Daily chemical dosing continued during the 5-day cohabitation. This second group of females was killed after 19 days of treatment and the number of live and dead fetuses and implantation sites was recorded. After 17 days of dosing, male mice were killed and the reproductive system evaluated by organ weights, total epididymal sperm counts and motility, and testicular histology. All four chemicals tested, boric acid, ethylene glycol, ethylene glycol monomethyl ether, and theophylline, were found to be toxic to development or reproduction when tested previously by conventional developmental toxicity or continuous breeding protocols. This short-term (21 day) design correctly identified three of these four chemicals as reproductive and developmental toxicants and distinguished the potent toxicants from the less effective compounds. This design can be used to prioritize chemicals for further study, or to delineate the relative toxicities of structurally related chemicals, and to identify the proper dose range for subsequent toxicity studies.

Animals↗