X-ray reflectivity measurement of interdiffusion in metallic multilayers during rapid heating.

A technique for measuring interdiffusion in multilayer materials during rapid heating using X-ray reflectivity is described. In this technique the sample is bent to achieve a range of incident angles simultaneously, and the scattered intensity is recorded on a fast high-dynamic-range mixed-mode pixel array detector. Heating of the multilayer is achieved by electrical resistive heating of the silicon substrate, monitored by an infrared pyrometer. As an example, reflectivity data from Al/Ni heated at rates up to 200 K s-1 are presented. At short times the interdiffusion coefficient can be determined from the rate of decay of the reflectivity peaks, and it is shown that the activation energy for interdiffusion is consistent with a grain boundary diffusion mechanism. At longer times the simple analysis no longer applies because the evolution of the reflectivity pattern is complicated by other processes, such as nucleation and growth of intermetallic phases.

Implementation of minimally invasive and objective humane endpoints in the study of murine Plasmodium infections.

SUMMARY Defining appropriate and objective endpoints for animal research can be difficult. Previously we evaluated and implemented a body temperature (BT) of <32 °C as an endpoint for experimental cerebral malaria (ECM) and were interested in a similar endpoint for a model of severe malarial anaemia (SMA). Furthermore, we investigate the potential of a minimally invasive, non-contact infrared thermometer for repeated BT measurement. ECM was induced with Plasmodium berghei ANKA infection in C57Bl/6 mice. SMA was induced with Plasmodium chabaudi AS infection in A/J mice. Our previous published endpoint was applied in ECM and 30 °C was pre-determined as the lowest permitted limit for termination in SMA according to consultation with the Danish Animal Inspectorate. Infrared thermometer was compared with the rectal probe after cervical dislocation, ECM and SMA. Linear regression analysis of rectal versus infrared thermometry: cervical dislocation: Pearson R = 0·99, R 2 = 0·98, slope = 1·01, y-intercept = 0·55; ECM: 0·99, 0·98, 1·06, -2·4; and SMA: 0·98, 0·97, 1·14, -5·6. Implementation of the 30 °C endpoint captured all lethal infections. However, some animals with BT below 30 °C were not deemed clinically moribund. This study supports repeated measurement infrared thermometry. A humane endpoint of 30 °C was sensitive in capturing terminal animals but might overestimate lethality in this SMA model.

Modification of the electro-optical properties of the B1 liquid-crystal phase using a rodlike liquid-crystal dopant.

It has recently been observed that on application of an alternating electric field, the B1 liquid-crystalline phase may be induced to form a switchable phase. This induced phase has been shown to have an almost thresholdless dielectric response. In this paper we examine this E-field-induced transition as a function of temperature both with and without a ferroelectric liquid-crystal dopant. Although the aim of this experiment was to enhance and stabilize the field-induced phase on addition of dopant, we find that the opposite is the case. The ferroelectric dopant actually increases the threshold E-fields required for transformation to the switching phase and at just 5 wt % dopant a smectic-A phase is formed. Addition of the dopant also acts against the low-field switching and significantly speeds relaxation back to the B1 phase on field removal. In addition we find that the field-induced phase experiences a slower crystallization. We use polarized optical microscopy, differential scanning calorimetry, and x-ray scattering experiments to characterize the physical properties of the mixtures.

Investigation into liquid crystalline smectic-C* subphase stability using chiral and achiral dopants.

In this paper we investigate the influence of chiral and achiral dopants on a chiral smectic liquid crystal material, which exhibits a rich phase sequence, including the antiferroelectric phase and the three-layer intermediate smectic (SmC*FI1) phase. Using polarized optical microscopy, differential scanning calorimetry, and x-ray diffraction we find that small amounts of achiral dopant have the ability to significantly broaden the SmC*FI1 phase, whereas an oppositely-handed dopant (otherwise identical to the host material) destabilizes the phase. This work clearly indicates that bulk chirality strongly influences SmC*FI1 phase formation and that steric effects also play an important role. Interestingly, addition of the shorter achiral molecule (8CB) was observed to increase the smectic layer spacing, most likely by suppressing interdigitation of alkyl chains between adjacent smectic layers. Control of the SmC*FI1 phase width using mixtures in this way is clearly important for effective phase characterization, and could potentially lead to commercially viable materials with a stable SmC*FI1 phase over a large temperature range.

Asymmetric synthesis of alpha-branched primary amines on solid support via novel hydrazine resins.

[reaction: see text]. Two novel chiral hydrazine resins for asymmetric solid-phase synthesis have been developed. The enantiopure beta-methoxyamino auxiliaries, derived from trans-4-hydroxy-(S)-proline and (R)-leucine, were attached to Merrifield resin and transformed into their corresponding hydrazines. Immobilization of various aldehydes, followed by 1,2-addition of organolithium reagents to the resulting enantiopure hydrazones and reductive cleavage from the solid support, furnished alpha-branched amines, which were isolated as their corresponding amides in good overall yields and enantiomeric excesses of up to 86%.

Impact of oral bases on aluminum absorption.

Control of hyperphosphatemia in renal failure is often difficult to achieve. Although calcium-containing phosphate binders have become the preferred phosphate binders, many patients require the addition of an aluminum-containing phosphate binder (APB). Enhanced aluminum absorption has been noted when APBs are administered with citrate-containing products such as citrate/citric acid solution (CCA). Alternative phosphate binders such as calcium acetate may also increase aluminum absorption. This study investigated the effect of CCAs on aluminum absorption when aluminum antacids (APBs) were administered concurrently and 2 hours apart. The effects of the alternative alkalinizing agent sodium bicarbonate and the alternate phosphate binding agent calcium acetate on aluminum absorption were also studied. During five 2-day phases, ten normal volunteers randomly received three times daily with standardized meals aluminum hydroxide alone and concurrently with NaHCO3, calcium acetate, CCA, or with CCA 2 hours postprandially. Twenty-four hour urines were collected on the second day of each phase and aluminum excretion was determined using inductively coupled plasma emission spectroscopy. Urine aluminum excretion was statistically significantly (P <.005) elevated in subjects receiving Al(OH)3 and CCA both with meals, 269.3 +/- 146.3 microg/d, and 2 hours after meals, 303.3 +/- 142.9 microg/d, compared with 79.2 +/- 52.0 microg/d during treatment with Al(OH)3 alone. Administration of CCA 2 hours after APB does not permit the safe use of these agents concurrently. Concomitant administration of sodium bicarbonate and calcium acetate with APBs appears to be safe, as aluminum absorption was not affected.

Electrochemical detection of thiols with a coenzyme pyrroloquinoline quinone modified electrode.

A modified electrode sensor for the detection of thiols is described. The sensor was constructed by incorporation of the coenzyme pyrroloquinoline quinone (PQQ) into a polypyrrole (PPy) film on a glassy carbon electrode substrate by the electropolymerization of pyrrole in the presence of PQQ. The electrochemical properties of entrapped PQQ in the PPy film were influenced by the applied potential during electropolymerization and by film thickness, both of which were optimized to yield a stable and reproducible response for entrapped PQQ. The PQQ/ PPy sensor was utilized for the amperometric detection of cysteine, homocysteine, penicillamine, N-acetylcysteine, and glutathione. The response for each thiol in pH 8.42 borate buffer was found to be linear with detection limits (S/N = 3) ranging from 13.2 microM for glutathione to 63.7 nM for cysteine with sensitivities of 0.023 nA/microM and 4.71 nA/microM, respectively. The response and detection limits were found to be sensitive to the nature of the thiol and the solution pH. Furthermore, in the presence of dopamine, ascorbic acid, or uric acid, the pH-dependent redox potential of the PQQ catalyst allows tuning of the detection potential to enhance the selectivity for thiols over these potential electroactive interferences.

Separation of the enzyme cofactor pyrroloquinoline quinone and three isomeric analogues by capillary electrophoresis with ion-pairing media.

The enzyme cofactor pyrroloquinoline quinone (PQQ) was successfully separated from three closely related isomeric analogues by capillary electrophoresis with ultraviolet detection. Rapid and efficient separation of all four negatively charged isomers with baseline resolution was achieved by the addition of low concentrations (1-5 mM) of short chain tetraalkylammonium (TAA) salts to the capillary buffer. The TAA cations act as ion-pairing agents and promote differential migration of the isomers with only a minimal reduction in the electroosmotic flow. The effects of the TAA salt concentration and the alkyl chain length were examined. Detection limits of PQQ and its isomers were in the range of 7-15 microM with mass detection limits of 98-210 fmol.

6-Hydroxycatecholine, a choline-mimicking analogue of the selective neurotoxin, 6-hydroxydopamine.

The synthesis and properties of 6-hydroxy-N,N-dimethylepinephrine (6-hydroxy-catecholine) are reported. This agent is intended for use as a selective presynaptic cholinotoxin and is based on previously reported neurotoxins of the same type. 6-hydroxycatecholine is a close structural analogue of the catecholaminergic neurotoxin 6-hydroxydopamine, and is expected both to be selectively reactive at cholinergic sites and to undergo less vigorous and potentially more selective inactivating reactions. It is also possible that in specific dementia-inducing pathologies, 6-hydroxycatecholine could be formed endogenously.

Dual-electrode detection for capillary electrophoresis/electrochemistry.

The extremely low sample volumes required for capillary electrophoresis and the high sensitivity and selectivity of electrochemical detection make capillary electrophoresis/electrochemistry (CEEC) a very useful method for bioanalysis. In this paper, two types of dual-electrode detectors for CEEC are described. The first employs a ring-disk microelectrode placed in a wall-jet configuration and is used for the selective detection of substances undergoing chemically reversible oxidations. Collection efficiencies obtained for catecholamines with this configuration were between 25 and 35%. The second electrode design consists of two adjacent carbon fibers embedded in an epoxy matrix and is analogous to the parallel dual-electrode configuration used in liquid chromatography/electrochemistry. This configuration can be used to confirm peak identity and purity by operating the electrodes at two different potentials. Alternatively, it is possible to perform simultaneous oxidative and reductive electrochemical detection.

Chemical vapor deposition fabrication and characterization of silica-coated carbon fiber ultramicroelectrodes.

Carbon fiber disk ultramicroelectrodes (UMEs) with well-defined geometries were prepared by chemical vapor deposition techniques. Transparent silica films with thicknesses from 1 to 600 microns were deposited on the cylindrical length of 5 and 10 microns carbon fibers from a SiCl4, H2, and O2 ternary precursor system at 850-1150 degrees C or sequential deposition from Si(OEt)4 as a single source precursor at 700 degrees C followed by the SiCl4, H2, and O2 precursor system. Film thickness, film adhesion to the fiber substrate, and the overall dimensions of the silica-coated carbon fiber were studied and found to be a function of the precursor system, precursor concentrations, fiber diameter, deposition time, and fiber temperature. The silica films were found to be free of microcracks and characterized by a quality seal between the carbon fiber and the coating. As a result, the silica-coated disk UME exhibits an excellent electrochemical response without the need to use an epoxy sealant at the electrode tip. Furthermore, the deposition of hard and inert ceramic materials imparts durability to fragile carbon fibers and facilitates the handling of UMEs in microenvironments. Finally, the advantage of concentric deposition about the fibers to produce a disk UME in the center of an insulating plane was used to examine the effect of the thickness of the insulating coating on the limiting current response.

High performance liquid chromatographic separation and pH-dependent electrochemical properties of pyrroloquinoline quinone and three closely related isomeric analogues.

Pyrroloquinoline quinone 1 (PQQ, methoxatin, 4,5-dihydro-4,5-dioxo-1H-pyrrolo[2,3-f]quinoline-2,7,9-tricarboxylic acid) is a novel coenzyme. Closely related isomeric analogues of PQQ may be formed in nature and may possess PQQ-like functions. Here, the electrochemical pH-dependent properties and the high performance liquid chromatographic (HPLC) separation protocols are reported for three major isomeric PQQ analogues likely to be formed in nature. The electrochemical data (cyclic voltammetry) provided the pKas of the five ionizable groups in each isomeric PQQ. The HPLC protocols provide the means of investigating the formation of PQQ and the PQQ isomers reported here during the normal turnover of eukaryotic amine oxidases containing integral topaquinone residues, as well as from direct enzyme-mediated reactions which remain to be characterized.

Carbon ring-disk ultramicroelectrodes.

Novel ring-disk ultramicroelectrodes (RD-UMEs) with analytical tip diameters as small as 25-30 microns were fabricated. Carbon RD-UMEs were reproducibly prepared by the chemical vapor deposition of alternating concentric layers of silica and carbon on resistively heated 10 microns carbon fibers. High-quality films with excellent adhesion at the interfaces between the carbon and silica layers were shown by electrochemical and scanning electron microscopy measurements. Electrochemical measurements of a solution of 1.0 mM ferrocene with 200 mM LiClO4 in CH3CN were used to characterize the single- and dual-electrode response lf the RD-UME. The electrochemical responses of the ring and the disk are sigmoidal in shape and indicated that radial diffusion is the primary mode of mass transport at each electrode at slow scan rates. Diffusion-controlled generation-collection experiments showed that the concentric dual-electrode configuration exhibits high collection efficiencies at the ring electrode with a 2-5 microns separation between electrodes and a 2-4 microns ring thickness. Close proximity of the ring and disk electrodes led to enhanced detection sensitivity due to back diffusion of regenerated molecules of a reversible redox couple from the collector to the generator electrode.

Mechanism of action of the redox affinity reagent [(trimethylammonio)methyl]catechol.

The synthesis of 4- and 5-hydroxy-3-[(trimethylammonio)methyl]catechol (4- and 5-HTMC) was carried out to examine their proposed involvement as intermediates in the spontaneous redox-dependent half-of-sites inactivation of neurotoxin binding sites in the nicotinic acetylcholine receptor (nAcChR) mediated by the parent compound 3-[(trimethylammonio)methyl]catechol (TMC) [Nickoloff et al. (1985) Biochemistry 24, 999-1007]. Oxidation of 4- and 5-HTMC occurred with sodium periodate with facile conversion to the corresponding p-quinones which were intercepted with thiols and cyclopentadiene. Both 4- and 5-HTMC inactivated neurotoxin binding in the nAcChR in a time course and over a concentration range consistent with their involvement as intermediates in the TMC redox-dependent inactivation of neurotoxin ([125I]-alpha-bungarotoxin) binding sites. Rapid concentration-dependent inactivation of neurotoxin sites occurred over a 10-1000 microM range and was resistant to further inactivation after 50% loss of available toxin binding sites on the nAcChR. Both 4- and 5-HTMC inactivated nAcChR neurotoxin sites much more rapidly and efficiently than was observed previously with TMC. The apparent binding constants for 4- and 5-HTMC with the nAcChR, calculated from their concentration-dependent inactivation behavior toward toxin binding sites, were Kd = 224 +/- 98 and 39 +/- 17 microM, respectively. The observed results and the redox potentials (vs Ag/AgCl reference electrode) measured by cyclic voltammetry at pH 1.8 for TMC (719 mV) and the 4- and 5-HTMC derivatives (519 and 443 mV, respectively) supported the previously proposed mechanism for inactivation of the nAcChR by TMC.(ABSTRACT TRUNCATED AT 250 WORDS)

[Formation and degradation of triademinol after the use of triadimefon in a wheat monoculture]. / Bildung und Abbau von Triadimenol nach Anwendung von Triadimefon in einer Weizenmonokultur.

During two vegetation periods, the behaviour of the triadimefon metabolite, triadimenol, in different plant parts of winter wheat and in soil was investigated. The fungicide Bayleton DF (triadimefon + captafol) was applied at the beginning of earing. Different ratios of triadimenol-A/-B were found in individual plant parts. Triadimenol-A predominated in the two uppermost leaves, and triadimenol-B in the roots and in the soil. No residues of triadimenol were found in grain at harvest time (detection limit 0.01 mg/kg).

[Behavior of captafol residues after prolonged application in a wheat monoculture]. / Abbauverhalten von Captafol nach langjähriger Anwendung in einer Weizenmonokultur.

In a 4-year study, the behaviour of the residues of the fungicide captafol in different plant parts of winter wheat and in the soil was investigated. The fungicide Bayleton DF (captafol + triadimefon) was applied at the beginning of earing. Captafol residues clearly showed a high dependence on weather conditions in all examined aerial parts of the plant. After a 9-year application of captafol, there was no evidence of an enrichment of residues in the soil. At harvest, captafol residues in grains always were below the maximum residue limits of 0.5 mg/kg established in West Germany.

Characterization of technetium radiopharmaceuticals by thin-layer spectroelectrochemistry.

Optically transparent thin-layer electrode (OTTLE) techniques have been used to characterize technetium complexes of biomedical importance. Different Tc oxidation states have been generated at the electrodes and absorption spectra determined. Small cell volumes allow the study of materials available in small quantities, and a flow cell configuration has also been used. Heart and bone imaging agents have been studied.

Interactions of methylestrogens with cytoplasmic and nuclear estrogen receptors in rat pituitary gland, hypothalamus and uterus.

The in vitro affinities to cytoplasmic estrogen receptors of the methylestrogens 2-methylestradiol-17 beta, 4-methylestradiol-17 beta and 4-hydroxy-2-methylestradiol-17 beta, which are useful probes to test the biological importance of 2- or 4-hydroxylation of estradiol-17 beta (catecholestrogen formation), have been determined in hypothalamic, pituitary and uterine tissue of the ovariectomized rat. Moreover, the in vivo capacity of these compounds to translocate estrogen receptors into the cell nucleus of pituitary and uterine tissue has been studied. Methylestrogens exhibited estrogen receptor affinities which were not significantly different from the binding affinity of estradiol-17 beta. When given at a high dose (100 micrograms/animal) their nuclear translocation capacity was equal (4-methylestradiol-17 beta) or even higher (2-methylestradiol-17 beta) than that of estradiol-17 beta. However, at a low dose (5 micrograms/animal) 4-methylestradiol was completely ineffective in both the pituitary gland and the uterus, and 2-methylestradiol-17 beta was less potent than estradiol-17 beta in the pituitary gland.