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Boron isotopes as an artificial tracer.

A field study was conducted using a combination of intrinsic and artificial tracers to estimate travel times and dilution during transport of infiltrate from a reclaimed water infiltration basin to nearby monitoring wells. A major study objective was to validate boric acid enriched in (10)B as an artificial tracer. Basin 10E at the Rio Hondo Spreading Grounds in Whittier, California, was the site of the test. The basin normally receives a mixture of treated municipal waste water, purchased State Project water, and local runoff from the San Gabriel River. Approximately 3.5 kg of (10)B-enriched boric acid was dispersed among 2.05 x 10(5) m(3) of basin water to initiate the experiment. The resultant median delta(11)B in the infiltration basin was -71 per thousand. Prior to tracer addition, the basin water had an intrinsic delta(11)B of +2 per thousand. Local monitoring wells that were used to assess travel times had delta(11)B values of +5 per thousand and +8 per thousand at the time of tracer addition. Analytic results supported an assumption that boron is conserved during ground water transport and that boron enriched in (10)B is a useful artificial tracer. Several intrinsic tracers were used to reinforce the boric acid tracer findings. These included stable isotopes of oxygen (delta(18)O) and hydrogen (deltaD), sulfate concentration, and the boron to chloride ratio. Xenon isotopes, (136)Xe and (124)Xe, also supported boron isotope results. Xenon isotopes were added to the recharge basin as dissolved gases by investigators from the Lawrence Livermore National Laboratory.

Boron↗

Evaluation of preservation methods and solid media suitable for recovery of Ureaplasma urealyticum from transported urine specimens.

Urine specimens to be tested for Ureaplasma urealyticum (T-strain mycoplasma) must often be transported to a central laboratory for identification. To examine survival of these organisms over 2 days of transport, fresh urine samples were tested for the presence of ureaplasmas and then divided into four groups: (i) fresh, (ii) frozen, (iii) preserved with 1% (wt/vol) boric acid at room temperature, and (iv) preserved with 10% dimethyl sulfoxide and frozen. The samples were cultured on both A7 and New York City solid media, and the estimates of survival were compared to that of fresh urine. Less than 10% survival of Ureaplasma was observed in 11 of 14 specimens stored frozen for 2 days without preservation; six specimens lost all organisms. Specimens containing either dimethyl sulfoxide or boric acid showed higher survival rates, although neither method consistently approached the full recovery of the T-strains found with the fresh urine. Ureaplasmas from fresh specimens grew well on both New York City and A7 media; however, the New York City medium proved superior for those preserved with boric acid and for urine samples containing few ureaplasmas. These results indicate that preservation of samples does increase the yield of U. urealyticum from urine samples delayed in transit.

Bacteriological Techniques↗

Simultaneous determination of HIV-protease inhibitors lamivudine and zidovudine in pharmaceutical formulations by micellar electrokinetic chromatography.

A micellar electrokinetic chromatographic (MEKC) method for the simultaneous separation and determination of lamivudine (LMV) and zidovudine (ZDV) in pharmaceutical formulation has been developed. Factors that affect the separation, such as buffer pH, surfactant concentration (sodium dodecyl sulfate, SDS), organic solvents and applied voltage were optimized. Buffer consisting of 12.5 mM sodium tetraborate decahydrate and 15 mM boric acid adjusted at pH 10.8, containing 90 mM SDS and 5% (v/v) acetonitrile (ACN) was found to be suitable for the separation of the drugs. p-Aminobenzoic acid (PABA) was used as internal standard (I.S.). Detection of analytes and I.S. was performed at a wavelength of 210 nm. It was observed that both the drugs and I.S. were migrated within 20 min at the applied voltage of +10 kV. Validation of the method was performed in terms of linearity, accuracy, precision, limit of detection (LOD) and quantification (LOQ). An excellent linearity was obtained in the concentration range 10-80 microg/ml for LMV and 10-100 microg/ml for ZDV. The detection limits for LMV and ZDV were found to be 2.5 and 2.0 microg/ml, respectively. The optimized method was applied to the simultaneous determination of LMV and ZDV in pharmaceutical formulation and human plasma (spiked) samples. Recovery of both the drugs in tablet dosage form and spiked drugs in plasma were > or =99.72% (relative standard deviation (R.S.D.)< or =1.84%) and > or =80.4% (R.S.D.< or =5.4%), respectively. In the electropherogram no interfering peaks were observed in the region of analytes and I.S. due to inactive ingredients in the tablets and matrices in plasma.

4-Aminobenzoic Acid↗

Kinetic analysis of boron transport in Chara.

The permeability of biological membranes to boric acid was investigated using the giant internodal cells of the charophyte alga Chara corallina (Klein ex Will. Esk. R.D. Wood). The advantage of this system is that it is possible to distinguish between membrane transport of boron (B) and complexing of B by plant cell walls. Influx of B was found to be rapid, with equilibrium between the intracellular and extracellular phases being established after approximately 24 h when the external concentration was 50 microM. The intracellular concentration at equilibrium was 55 microM, which is consistent with passive distribution of B across the membrane along with a small amount of internal complexation. Efflux of B occurred with a similar half-time to influx, approximately 3 h, which indicates that the intracellular B was not tightly complexed. The concentration dependence of short-term influx measured with 10B-enriched boric acid was biphasic. This was tentatively attributed to the operation of two separate transport systems, a facilitated system that saturates at 5 microM, and a linear component due to simple diffusion of B through the membrane. Vmax and Km for the facilitated transport system were 135 pmol m(-2) s(-1) and 2 microM, respectively. The permeability coefficient for boric acid in the Chara plasmalemma estimated from the slope of the linear influx component was 4.4 x 10(-7) cm s(-1) which is an order of magnitude lower than computed from the ether:water partition coefficient for B.

Biological Transport↗

Analysis of selected constituents in methanolic extracts of Hypericum perforatum collected in different localities by capillary ITP-CZE.

The on-line combination of CZE with capillary ITP (ITP-CZE) was used for the separation and quantification of selected flavonoids and phenolic acids in Hypericum perforatum leaves and flowers collected in six different localities in Slovakia. The leading electrolyte in the ITP preseparation step was 10 mM HCl with Tris as counterion (pH* 7.2). The terminating electrolyte was 50 mM boric acid of pH* 8.2 (adjusted with barium hydroxide). The BGE in the electrophoretic step contained 25 mM beta-hydroxy-4-morpholinopropanesulfonic acid (MOPSO), 50 mM Tris, 65 mM boric acid, pH* 8.3. The content of methanol in all electrolytes was 20% v/v. The total time of the analysis (including the preseparation step) was approximately 35 min. The rectilinear calibration ranges were between 0.125 and 5.0 microg/mL with kaempferol as internal standard. The correlation coefficients ranged between 0.9912 (for quercitrin and chlorogenic acid) and 0.9988 (for isoquercitrin). The RSD values are between 0.86 and 7.78% (n = 6) when determining rutin and quercetin (4 microg/mL). The optimized method was employed for the assay of flavonoids in medicinal plant extract of different collections of Hypericum perforatum haulm. The variability of the content of the active components depending on the place of collection was confirmed.

Boric Acids↗

A study of three preparations in the treatment of otitis externa.

Twenty-seven patients with an initial diagnosis of otitis externa, untreated for the previous 2 weeks, were treated with weekly aural suction toilet and a choice of three topical drops. Two contained combinations of antibiotics and steroids, one of which also contained an anti-fungal agent. The third was boric acid in spirit. The patients were assessed at weekly intervals until a remission was established. From this study it would appear that antibiotic-based drops confer no real advantage over boric acid in spirit.

Administration, Topical↗

Perak, Malaysia, mass poisoning. Tale of the Nine Emperor Gods and rat tail noodles.

In October 1988, a series of food poisoning cases occurred in the State of Perak in Malaysia. Most of the victims were children. Ultimately 13 children between the ages of 2.5 and 11 years died. Epidemiological investigations showed that the probable source of the poison was Loh See Fun, a noodles in the shape of a rat's tail. All the deceased ate the noodles from one supplier. Clinical and pathological findings were similar in each case. Postmortem examination was performed in 11 cases. Toxicological examination on organs in 10 cases showed a high concentration of aflatoxin in tissues of the deceased. High levels of boric acid were excreted from most of the victims. Histological examination of the liver in these cases showed necrotic changes found in aflatoxin poisoning. Combination of the epidemiological, clinical, toxicological, and pathological findings pointed to the fact that there was a common toxin or toxins responsible for the deaths. These were thought to be a combination of boric acid and aflatoxin.

Boric Acids↗

Reinvestigation of positional distribution of fatty acids in docosahexaenoic acid-rich fish oil triacyl-sn-glycerols.

Positional distribution of fatty acids in triacyl-sn-glycerols of docosahexaenoic acid (DHA)-rich tuna orbital and bonito head oils has been reanalyzed by a method based on chromatographic separation of isomeric and enantiomeric monoacyl-sn-glycerol (MAG) derivatives. When boric acid thin-layer chromatography (TLC) was used for separation of 1(3)- and 2-MAG analytical intermediates, the stereospecific analysis showed the preferential association of DHA to the sn-2 position followed by the sn-3 position. This distribution pattern differed from that obtained by silicic acid TLC of their bis-3,5-dinitrophenylurethane (DNPU) derivatives. Reversed-phase high-performance liquid chromatography elution profiles of 1(3)- and 2-MAG intermediates revealed that 1(3)- and 2-MAG made up of both short- and long-chain lengths cannot be clearly resolved by TLC after preparation of the DNPU derivatives. The 1(3)- and 2-MAG must be resolved by boric acid TLC prior to derivatization.

Animals↗

Evaluation of self-taken samples for the presence of genital Chlamydia trachomatis infection in women using the ligase chain reaction assay.

The aim of this study was to evaluate the sensitivity and acceptability of self-taken vulval-introital (VI) samples, first-catch urine (FCU) samples and clinician-obtained cervical samples for the presence of genital Chlamydia trachomatis infections in women using the ligase chain reaction (LCR) assay. One hundred and four patients were enrolled, of whom 54 patients had chlamydial DNA in at least one of the samples tested. The sensitivity of the cervical sample was 96.3%, vulval-introital sample in LCR buffer 92.6%, vulval-introital swab collected dry 88.9%, FCU stored at +2-8 degrees C 81.5%, FCU stored at room temperature 77.8% and FCU stored with 2% w/v boric acid at room temperature 87.0%. Self-taken vulval-introital LCR samples were shown to be an acceptable alternative to a clinician-obtained LCR sample. The addition of boric acid may overcome the need for a continuous cold chain for FCU samples.

Adolescent↗

Determination of diterpenoid triepoxides in Tripterygium wilfordii by micellar electrokinetic capillary chromatography.

A micellar electrokinetic capillary chromatographic (MEKC) method has been established for the identification and determination of diterpenoid triepoxides in the Chinese herb Tripterygium wilfordii Hook F. and its preparations. Studies of the influence of boric acid and borax buffer concentration and pH, and of sodium dodecylsulphate (SDS) concentration have been carried out, and the optimum separation for the triepoxides was achieved using 20 mM boric acid and 10 mM borax with 20 mM SDS as the running buffer. MEKC was found to exhibit good accuracy, precision and repeatability. The sensitivity of the assay was sufficient to monitor the three active components in T. wilfordii and its preparations.

Borates↗

[Effect of colchicine and other polyploidogenic factors in contact with the deep vegetative mycelium of the fungus Trichothecium roseum--producer of the antibiotic trichothecin and proteolytic enzymes].

Colchicine, boric acid, and camphor were used to produce viable cultures of Trichothecium roseum actively producing the antibiotic, by treating the submerged vegetative mycelium of the fungus with these compounds. Colchicine (0.5--1.0%), boric acid (0.25--2.00%), and camphor (0.25--1.50%) were added to the nutrient medium before it was inoculated with conidia or vegetative mycelium; later, the submerged mycelium was transferred to slanted agar. The cultures were characterized by a high number of the nuclei in the conidia, a low ratio of the plasma to the nuclei, a high rate of growth, and a high antibiotic activity. In sharply varying conditions of nutrition, such polynuclear forms had an adaptive advantage as compared to the parent forms. The polynuclear cultures are regarded to be biologically equal to polyploid forms.

Anti-Bacterial Agents↗

Separation and determination of sorbitol and xylitol in multi-component pharmaceutical formulations by capillary isotachophoresis.

Pharmaceutically important polyhydric alcohols sorbitol (SO) and xylitol (XY) are efficiently separated and determined by analytical capillary isotachophoresis (ITP) with conductometric detection. The on-column complex-formation equilibria between the polyols and boric acid are utilized--the terminating borate ion acts as the complexing agent. The ITP operational system used consists of 10 mM HCl + 20 mM imidazole (LE, pH 7.0) and 20 mM boric acid (TE, pH 8.0). The effective mobilities of the borated SO and XY are 8.3 x 10(-9) m2 V-1 s-1 and 7.4 x 10(-9) m2 V-1 s-1, respectively. The ITP analysis is performed with the driving and detection currents of 50 microA (for 700 s) and 20 microA, respectively. The calibration graphs are rectilinear in the range 25-250 mg l-1 of SO and 50 to 500 mg l-1 of XY. The method is applied to the simultaneous assay of SO and XY in three mass-produced multi-component infusion solutions. Favourable values of the method validation parameters obtained confirm the suitability of the proposed ITP method for the quality control of pharmaceuticals.

Boric Acids↗

The distribution of components in the different size fractions of granules prepared from binary mixtures.

Boric acid, sulphanilamide and citric acid have been mixed separately with lactose and then granulated by massing and screening. The granules have been fractionated by sieving and each fraction has been analysed for lactose content. The effect of premixing time, massing time, binder volume and ratio of components on the distribution of lactose between size fractions of granules prepared from lactose: boric acid mixtures has been investigated. Uneven distribution of lactose has been found for all blends examined. There is a premixing time and massing time that gives the optimum distribution of lactose for any given blend and binder volume. Increased binder volume in some cases improves granule uniformity. The proportion of lactose in the blend has a major effect on the distribution of this component in the granules, as does the particle size of the lactose. Granules prepared from blends of lactose with sulphanilamide and with citric acid were also examined for lactose distribution.

Boric Acids↗

Novel extraction of German cockroach (Dictyoptera: Blattellidae) fecal pellets enhances efficacy of spray formulation insecticides.

Methanol extracts of German cockroach, Blattella germanica (L.), fecal pellets have limited use as pheromone attractants in the urban environment because of their unpleasant color and odor. To eliminate these characteristics, a novel aqueous extract of German cockroach feces was formulated. Fetal material was extracted with methylene chloride and mixed with water. The aqueous phase of the extract was colorless and relatively odorless compared with methanol fecal extracts. Aqueous extract was bioassayed and compared with methanol extracts for aggregation activity. The efficacy of chlorpyrifos and boric acid formulations was tested with and without the addition of fecal extracts. In tests with chlorpyrifos, the addition of the aqueous extract produced significantly greater mortality in young nymphs than either chlorpyrifos alone or chlorpyrifos + methanol extract. There were no differences in mortality between the 2 extract + boric acid treatments during the test period, and both enhanced mortality.

Animals↗

The granulation of binary mixtures.

Granules have been prepared from blends of lactose and boric acid by the massing and screening method and their properties have been compared with those of granules from the individual materials, granulated separately. Increasing the volume of binder solution used produced granules stronger and of greater mean size in all blends studied. The largest and strongest granules, for any given binder solution volume and massing time, were obtained from a blend, as were the granules with the minimum pore size. The effect of massing time on granule properties was generally similar to those already reported for single component systems; prolonged massing in some blend led to a reduction in mean granule size. Variation in pre-mixing time produced no significant change in the physical properties of granules prepared from a lactose: boric acid blend.

Boric Acids↗

Determination of urinary free noradrenaline by reversed-phase high-performance liquid chromatography with on-line extraction and fluorescence derivatization.

A fully automated reversed-phase high-performance liquid chromatographic method with fluorimetric detection for the determination of urinary free noradrenaline was developed. Urine samples, diluted with buffer were injected into a boric acid gel column (12 microns, TSK: 10 x 4.6 mm I.D.) without prior extraction. Urinary noradrenaline was simultaneously extracted and derivatized with an alkaline mobile phase of pH 11, containing o-phthalaldehyde and 2-mercaptoethanol, in a boric acid gel column. After switching columns, the fluorescent derivatized catecholamines were separated with an ODS-4PW column (TSK) and a mobile phase of pH 2 and the fluorescence was monitored with excitation at 340 and emission at 440 nm. The retention times of noradrenaline and dopamine were 11.0 and 14.2 min, respectively. The detection limits for noradrenaline and dopamine were 0.2 and 20 ng, respectively. This method has the advantages of not requiring preliminary extraction of urinary catecholamines, high sensitivity and stability of o-phthalaldehyde-derivatized catecholamines.

Boric Acids↗

Lyso platelet activating factor (LysoPAF) and its enantiomer. Total synthesis and carbon-13 NMR spectroscopy.

Described is a reaction sequence for the total synthesis of lyso platelet activating factor (lysoPAF; 1-O-alkyl-sn-glycero-3-phosphocholine) and its enantiomer. The procedure is versatile and yields optically pure isomers of defined chain length. The synthesis is equally suited for the preparation of lysoPAF analogues and its enantiomers with unsaturation in the long aliphatic chain. First, rac-1(3)-O-alkylglycerol is prepared by alkylation of rac-isopropylideneglycerol with alkyl methanesulfonate followed by acid-catalyzed removal of the ketal group. The primary hydroxy group of alkylglycerol is then protected by tritylation, the secondary hydroxy group is acylated, and the protective trityl group is removed under mild acidic conditions with boric acid on silicic acid, essentially without acyl migration. Condensation of the diradylglycerol with bromoethyl dichlorophosphate in diethyl ether, hydrolysis of the resulting chloride, and nucleophilic displacement of the bromine with trimethylamine gives rac-1-O-alkyl-2-acylglycero-3-phosphocholine in good overall yield. The racemic alkylacylglycerophosphocholine is finally treated with snake venom phospholipase A2 (Ophiophagus hannah) which affords 1-O-alkyl-sn-glycero-3-phosphocholine (lysoPAF) of natural configuration in optically pure form. The "unnatural" 3-O-alkyl-2-O-acyl-sn-glycero-1-phosphocholine enantiomer, which is not susceptible to phospholipase A2 cleavage, gives 3-O-alkyl-sn-glycero-1-phosphocholine upon deacylation with methanolic sodium hydroxide. Homogeneity and structure of the intermediates and final products were ascertained by carbon-13 nuclear magnetic resonance spectroscopy on monomeric solutions.

Acylation↗

Purification and characterization of urease isolated from the pathogenic fungus Coccidioides immitis.

Coccidioides immitis, the causative agent of San Joaquin Valley fever (coccidioidomycosis), produces a urease which has been suggested to contribute to the virulence of this fungal pathogen. Urease catalyzes the hydrolysis of urea and has been proposed to at least partly account for alkalinity of the microenvironment in which C. immitis grows due to the release of ammonia and ammonium ions. The C. immitis urease was purified to homogeneity (1048-fold) from the mycelial cytosol by chromatographic fractionation. The sequence of 12 N-terminal amino-acid residues of the purified, native polypeptide was identical to that predicted by the translated urease gene sequence which has been reported. The isolated enzyme exhibited a specific activity in the presence of urea of 1750 micromol min(-1) mg(-1) protein, has a native molecular mass of 450 kDa, revealed a Km for urea of 4.1 mM, had a pH optimum of 8.0 and is heat stable. Hydroxyurea, acetohydroxamic acid (AHA) and boric acid each inhibited activity of the purified enzyme. Urease activity was enhanced by the presence of 5-10 mM concentrations of Mg2+ or Mn2+, but inhibited by Li+, Ni2+, Cu2+ or Zn2+. The reversible urease inhibitor, AHA, blocked enzyme activity in the crude mycelial cytosolic fraction when added at a concentration of 10 mM. On the other hand, 10 mM AHA added to 4-day-old mycelial cultures only partially decreased the amount of ammonium detected in the culture medium. It is evident, therefore, that C. immitis urease activity does not account for the total amount of ammonia secreted during in vitro growth of the pathogen. Other metabolic sources of ammonia, which may also contribute to the virulence of C. immitis, are under investigation.

Amino Acid Sequence↗