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PubMed1993

A nitro radical anion formation from nifedipine: an electrochemical approach.

The cyclic voltammetric behaviour of nifedipine was studied. The addition of three aprotic solvents to nifedipine in an aqueous citrate buffer system was examined. Qualitatively they result in separation of the initial irreversible 4 electron reduction into two stages, the NO2/RNO2.- and RNO2.-, 4H/RNHOH, H2O couples, respectively. Particular attention was directed to the 1-electron RNO2/RNO2.- couple as measured by the cyclic voltammetric mode in mixed media. Analysis of the cyclic voltammetric response as a function of scan rate and non-aqueous solvent content yields information on the stability of the radical anion. The chemical forward reaction of the radical anion follows a second order kinetics with a stability constant of 1.1 x 10(-3) l mol-1 s-1 and a half-life time of 0.09 s for 1 mM of nifedipine in aqueous citrate buffer, pH 7.4/DMF; 50:50.

Electrochemistry↗
PubMed1995

Determination of ceftiofur and its desfuroylceftiofur-related metabolites in swine tissues by high-performance liquid chromatography.

An HPLC method was developed and validated for the determination of ceftiofur-related metabolites that have the potential to be microbiologically active in swine muscle, kidney, liver and fat. Its performance was evaluated against incurred-residue swine tissues. This method is based on the cleavage of the disulfide and/or thioester bonds between the metabolites and their conjugate sulfur containing moiety using dithioerythritol to yield desfuroylceftiofur, and further stabilization to desfuroylceftiofur acetamide. The limit of quantitation was 0.1 micrograms ceftiofur equivalents/g tissue. The assay is specific for ceftiofur-related metabolites when evaluated against commercially available antibiotics for swine.

Adipose Tissue↗
PubMed1992

N-(m-[125I]iodophenyl)maleimide: an agent for high yield radiolabeling of antibodies.

In an effort to radiolabel antibodies, N-(m-[125I]iodophenyl)maleimide (m-[125I]IPM) was prepared by the demetallation of an N-[m-tri-(n-butyl)stannylphenyl]maleimide intermediate. The unlabeled intermediate was synthesized in greater than or equal to 75% yield using a palladium catalyzed reaction of hexabutylditin with m-bromoaniline, followed by reaction with maleic anhydride and ring annulation. All products were confirmed by NMR and elemental analysis. Labeling with 125I was carried out in a biphasic mixture containing chloramine-T (radiochemical yield greater than or equal to 70%). Rabbit IgG modified with the heterobifunctional crosslinking agent N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP) and bovine serum albumin were conjugated with m-[125I]IPM (yield: 40 and 80%, respectively). In addition, m-[125I]IPM was conjugated to rabbit IgG subunits (HL) in 70% yield. The in vitro stability of the radiolabeled proteins in serum showed less than 1% deiodination over 24 h.

Animals↗
PubMed1992

Synthesis, characterization and biodistribution studies of a neutral-lipophilic Tc-99m N3S2 chelate.

Diaminedithiols (DADT) are known to form neutral-lipophilic complexes with 99mTc in aqueous solutions, where they are readily formed in high yields and demonstrate excellent stability. A new triaminedithiol (TADT) ligand was synthesized, characterized and shown to form a neutral-lipophilic 99mTc-chelate. The biodistribution of this 99mTc chelate in rats showed that its uptake in brain or heart following i.v. injection of the 99mTc chelate was low, but activity taken up was retained over a long period of time. The in vivo and in vitro properties of this chelate indicate the possibility that chemical modification of this TADT ligand may produce ligand systems that form 99mTc chelates with suitable diagnostic properties.

Animals↗
PubMed2005

Preparation and important functional properties of water-soluble chitosan produced through Maillard reaction.

The objective of this research was to improve the solubility of chitosan at neutral or basic pH using the Maillard-type reaction method. To prepare the water-soluble chitosans, various chitosans and saccharides were used under various operating conditions. Biological and physicochemical properties of the chitosan-saccharide derivatives were investigated as well. Results indicated that the solubility of modified chitosan is significantly greater than that of native chitosan, and the chitosan-maltose derivative remained soluble when the pH approached 10. Among chitosan-saccharide derivatives, the solubility of chitosan-fructose derivative was highest at 17.1 g/l. Considering yield, solubility and pH stability, the chitosan-glucosamine derivative was deemed the optimal water-soluble derivative. Compared with the acid-soluble chitosan, the chitosan-glucosamine derivative exhibited high chelating capacity for Zn(2+), Fe(2+) and Cu(2+) ions. Relatively high antibacterial activity against Escherichia coli and Staphylococcus aureus was noted for the chitosan-glucosamine derivative as compared with native chitosan. Results suggest that the water-soluble chitosan produced using the Maillard reaction may be a promising commercial substitute for acid-soluble chitosan.

Anti-Bacterial Agents↗
PubMed2006

Preparation of cyclotron-produced 186Re and comparison with reactor-produced 186Re and generator-produced 188Re for the labeling of bombesin.

The radioisotopes (186)Re and (188)Re have been extensively investigated for various forms of radiotherapy due to their useful and high-abundance beta particle emissions, low-abundance and imageable gamma-rays, and chemical resemblance to technetium. In addition, (188)Re is available in no-carrier-added (NCA) form from long lived W-188 generators, whereas (186)Re can be produced in large quantities from reactors, although not in NCA form. However, NCA (186)Re can be produced on a cyclotron by a (p,n) reaction on (186)W. The purpose of this study was to compare labeling of the peptide bombesin with these three forms of rhenium radioisotopes. Cyclotron-produced NCA (186)Re was separated radiochemically from enriched (186)W (96.9%) targets using high-purity methyl ethyl ketone (MEK). The resulting (186)Re-MEK was then loaded onto a small alumina column to separate the resulting NCA (186)Re from any remaining (186)W. The experimental levels of impurities associated with (186)Re at the end of the separation process were found to be 5.7 x 10(-6) Ci of (182)Re (0.57%, t(1/2) = 12.7 h) and 1.283 x 10(-5) Ci of (182m)Re (1.28%, t(1/2) = 2.67 days). The radionuclidic purity of the separated (186)Re was found to be 99.6%, whereas the chemical identity was determined by reversed phase high-performance liquid chromatography (RP-HPLC) to be perrhenate ((186)ReO(4)(-)). Generator-produced (188)ReO(4)(-) from a (188)W/(188)Re generator (Oak Ridge National Laboratory) and CA (186)ReO(4)(-) produced from a (185)Re(n,gamma)(186)Re reaction at the University of Missouri Research Reactor (MURR) were used for comparison with the NCA (186)Re in subsequent studies. N(3)S-5-Ava-BBN(7-14)NH(2) conjugates provide flexibility for designing (186,188)Re-labeled conjugates that retain high in vitro and in vivo specificity targeting of GRP receptor-expressing cells. This study showed that the N(3)S-5-Ava-BBN(7-14)NH(2) could be labeled with (186,188)Re following the preconjugation, postmetallation approach. The (186,188)Re(V)O-N(3)S-5-Ava-BBN(7-14)NH(2) complexes were found to form stable complexes following the reduction of perrhenate (Re(VII)O(4)(-)) with stannous chloride at room temperature, as verified by HPLC and stability studies. The radiolabeling yield was found to be >90%. The HPLC chromatograms of (186,188)Re-N(3)S-5-Ava-BBN(7-14)NH(2) complexes revealed two peaks for each conjugate, reflecting the presence of syn- and anti-isomers, which were resolvable by HPLC but re-isomerized on separation. The biodistribution studies showed that the compounds were excreted through the renal and hepatobiliary systems and demonstrated receptor-specific uptake with an average pancreas accumulation of 8.15% ID/g at 1 h postinjection. Administration of cold BBN effectively blocked pancreatic uptake and further reflects the high specificity this conjugate has for the GRP receptors. At low levels of radioactivity, radiolysis effects were not observed. Scale-up may or may not elicit this effect, particularly for the higher energy beta emitter (188)Re. The biodistribution studies demonstrated that the CA and NCA (186,188)Re conjugates behaved similarly, raising the question of whether NCA (186,188)Re is necessary for specific tumor receptor targeting.

Animals↗
PubMed2004

Improved formulations for existing CBPP vaccines--recommendations for change.

Contagious bovine pleuropneumonia (CBPP) is an economically important transboundary disease, widely present in sub-saharan Africa. Social, cultural and economic factors mean that effective vaccination is the only viable control policy at present. Unfortunately, contemporary live attenuated vaccines are reportedly of limited efficacy and have been unable to control recent outbreaks. Efforts to develop newer vaccine technologies are currently underway, although with little success to date. This review examines the prospects of success for such approaches, and argues that alternative strategies, based upon simple and inexpensive changes to current vaccines and protocols are likely to prove far more effective in the foreseeable future. Such changes include the use of HEPES-buffer systems and the inclusion of pH indicators in vaccine media, together with restrictions in the use of 1M MgSO4 as a vaccine diluent. These changes can increase vaccine yields 10-fold and stability several 100-fold, increase the ease of production, provide a significant level of end user-enforceable quality control, and ultimately produce a vaccine which should prove effective in the field immediately.

Africa↗
PubMed2002

Progress in the development and use of ferrate(VI) salt as an oxidant and coagulant for water and wastewater treatment.

This paper reviews the progress in preparing and using ferrate(VI) salt as an oxidant and coagulant for water and wastewater treatment. The literature revealed that due to its unique properties (viz. strong oxidizing potential and simultaneous generation of ferric coagulating species), ferrate(VI) salt can disinfect microorganisms, partially degrade and/or oxidise the organic and inorganic impurities, and remove suspended/colloidal particulate materials in a single dosing and mixing unit process. However, these findings have not yet lead to the full-scale application of ferrate(VI) in the water industry owing to difficulties associated with the relatively low yield of ferrate(VI), the instability of the chemical depending on its method of preparation, and the lack of adequate studies that have demonstrated its capabilities and advantages over existing water and wastewater treatment methods. Fundamental study is thus required to explore the new preparation methods focusing on increasing the production yield and product's stability and avoiding using hypochlorite or chlorine as the oxidant. Also, the application of ferrate(VI) in drinking water treatment has not been studied systematically and future work in this field is recommended.

Disinfection↗
PubMed1994

Resorbable dowels for osteosynthesis of noncongruent bone fragments.

Dowels, in contrast to screws, offer the great advantage in osteosynthesis that no pressure peaks occur by functional loading rectangular to the screw axis; consequently, there is no bone resorption and no loosening of the fixation because of the even contact of dowel and bony surface. However, dowels must be made of resorbable materials because they cannot be removed. We developed double-spreading dowels with a slotted tube whose outside is cylindric and the inside conical at both ends. Two conical screws in the core of the dowel moved toward each other cause symmetric expansion of the outer surface. Dowel material consisted of polylactic acid with an inherent viscosity > 7 dl/g (Resomer 214, Böhringer Ingelheim). A model simulating retromolar sagittal split osteotomy was chosen for experimental research because, in this case, noncongruent bone segments have to be osteosynthesized to regenerate in the sense of distance osteogenesis. Two segments of synthetic material were synthesized like proximal and distal segments after retromolar sagittal split osteotomy fixed by two or three dowels without coherent contact. The synthesized segments underwent a continuous jiggling stress test comparable with the physiologic stress of a jaw for about 4 weeks. Dowel fixation proved to be nearly as stable as position-screw fixation. Resorbable dowel fixation yields experimentally the required stability for the time needed for bony regeneration.

Analysis of Variance↗
PubMed2002

Pilot plant recovery of catheptic proteases from surimi wash water.

Recovery of bioactive compounds, such as proteolytic enzymes, from waste streams is a means to both recuperate value and reduce environmental pollution. Previously optimized lab-scale parameters for the recovery of a stable crude protease fraction from Pacific whiting (Merluccius productus) surimi wash water were tested using pilot plant equipment. Pretreatment of surimi wash water with 60 degrees C heat, acidification to pH 6, and centrifugation doubled ultrafiltration membrane flux and significantly improved protease purity by reducing a majority of the 35-205 kDa proteins. Concentrated crude protease obtained from wash water contained predominantly cathepsin L activity. Enzyme purity was increased about 100-fold, and yield was approximately 80%. Stability (frozen and freeze-dried protease) was maintained for 9 weeks at -80 degrees C. Freeze-dried preparations were also stable for 9 weeks at 4 and -15 degrees C. Successful application of pilot plant conditions allows for sufficient production of protease for further investigations into their applicability.

Animals↗
PubMed2002

A versatile planar QCM-based sensor design for nonlabeling biomolecule detection.

Despite high theoretical sensitivity, low-cost manufacture, and compactness potentially amenable to lab-on-a-chip use, practical hurdles have stymied the application of the quartz crystal microbalance (QCM) for aqueous applications such as detection of biomolecular interactions. The chief difficulty lies in achieving a sufficiently stable resonance signal in the presence of even minute fluctuations in hydrostatic pressure. In this work, we present a novel versatile planar sensor chip design (QCM chip) for a microliter-scale on-line biosensor. By sealing the quartz resonator along its edges to a flat, solid support, we provide uniform support for the crystal face not exposed to solvent, greatly decreasing deformation of the crystal resonator under hydrostatic pressure. Furthermore, this cassette design obviates the need for direct handling when exchanging the delicate quartz crystal in the flow cell. A prototype 27-MHz sensor signal exhibited very low noise over a range of flow rates up to 100 microL/min. In contrast, signals obtained from a conventional QCM sensor employing an O-ring-based holder were less stable and deteriorated even further with increasing flow rate. Additional control designs with intermediate amounts of unsupported undersurface yielded intermediate levels of stability, consistent with the interpretation that deformation of the crystal resonator under fluctuating hydraulic pressure is the chief source of noise. As a practical demonstration of the design's high effective sensitivity, we readily detected interaction between myoglobin and surface-bound antibody.

Animals↗
PubMed2003

Sol-gel-derived sensor materials that yield linear calibration plots, high sensitivity, and long-term stability.

Novel O2-sensing materials based on spin-coated n-octyltriethoxysilane (Octyl-triEOS)/tetraethylorthosilane (TEOS) composite xerogel films have been synthesized and investigated. These sensors are based on the O2 quenching of tris(4,7-diphenyl-1,10-phenanthroline)ruthenium(II) ([Ru(dpp)3]2+) sequestered within the xerogels. Scanning electron microscopy and luminescence measurements (steady state and time resolved) have been used to investigate the structure of these films and their analytical figures of merit and determine the underlying reasons for their observed performance. The results show that certain [Ru(dpp)3]2+-doped Octyl-triEOS/TEOS composites form uniform, crack-free xerogel films that can be used to construct high-sensitivity O2 sensors that have linear calibration curves and excellent long-term stability. For example, an 11-month-old sensor based on 50 mol % Octyl-triEOS exhibits more than 4-fold greater sensitivity in comparison to an equivalent sensor based on pure TEOS. Over an 11-month time period, the sensitivity of a pure TEOS-based sensor drops by more than 400% whereas a sensor based on 50 mol % Octyl-triEOS remains stable (RSD = 4%).

Biosensing Techniques↗
PubMed2003

Study of residual particle concentrations generated by the ultrasonic nebulization of deionized water stored in different container types.

A scanning mobility particle sizer has been used to quantify residual particle number and mass concentrations generated by ultrasonic nebulization of deionized (DI) water stored in a variety of bottles. High variability of residual particles was found not only between different bottle types but also between different bottles of the same type. Degradation of the water quality, quantified as increased residual mass and number concentrations as a function of time, occurred to varying degrees for water stored in different bottle types. Overall, glass bottles showed the highest residual particle concentrations and exhibited the poorest stability over time. After a storage period of 3 weeks, DI water stored in Pyrex bottles showed average increases in particle mass and number densities in the aerosol of over 250% and 60%, respectively. Total dissolved impurity levels in the water increased from 110 to 290 ng mL(-1) over the 3-week period. It is hypothesized that leaching from the bottle walls increases impurity levels in the water over time. Leaching was observed for both glass and polymer bottles. Contrary to this trend, residual particle concentrations from deionized water stored in Teflon bottles showed a net decrease during the measurement period. With respect to absolute residual particle concentrations and storage stability, a Teflon bottle yielded the best performance. Total residual particle mass and number densities for Teflon were less than a factor of 15% and 1%, respectively, as compared to residual particle levels observed for the Pyrex bottle. Absolute dissolved impurity levels in the water for the Teflon bottle decreased from 7.8 to 3.7 ng mL(-1) over the 4-week period.

Journal Article↗
PubMed1996

Synthesis and characterization of a caged receptor ligand suitable for chemical kinetic investigations of the glycine receptor in the 3-microseconds time domain.

Here we report the development and characterization of a new photolabile protecting group for the carboxyl group of neurotransmitters, 2-methoxy-5-nitrophenyl. The synthesis and characterization of a photolabile derivative of beta-alanine, caged beta-alanine, are described. beta-Alanine can activate the glycine receptor, a major inhibitory receptor in the mammalian central nervous system; the 2-methoxy-5-nitrophenyl derivative of beta-alanine combined with a laser-pulse photolysis method makes it possible to investigate the chemical kinetic mechanism of the receptor in the 3-microseconds time domain. The derivative is photolyzed by a laser pulse to release free beta-alanine within 3 microseconds and with a product quantum yield of 0.2. In aqueous solution in the dark and at neutral pH, the compound is more stable, by a factor of approximately of 25, than the analogous derivative of glycine [Ramesh, D., Wieboldt, R., Niu, L., Carpenter, B. K., & Hess, G. P. (1993) Proc. Natl. Acad. Sci. U.S.A. 90, 11074-11078]. 2-Methoxy-5-nitrophenyl-beta-alanine hydrolyzes in aqueous solution at neutral pH with a t1/2 of approximately 1.5 h. Neither the 2-methoxy-5-nitrophenyl-beta-alanine nor the 2-methoxy-5-nitrophenol photolysis side product activates, inhibits, or potentiates the response of glycine receptors in rat hippocampal neurons to glycine. Photolysis of 2-methoxy-5-nitrophenyl-beta-alanine by irradiation with a 600-ns laser pulse at 333 nm releases beta-alanine, which then activates glycine receptor-channels on neurons equilibrated with the caged compound, as detected by whole-cell current recording. Compared with the analogous derivative of glycine, in terms of quantum yield, photolysis rate, and stability, this new compound is not only a better candidate for use in chemical kinetic investigations of the glycine receptor, but can also be used in determining the location of glycine receptors in neuronal cells.

Animals↗
PubMedDate not supplied

Laccase-mediated Remazol Brilliant Blue R decolorization in a fixed-bed bioreactor.

A crude laccase mixture preparation from Pleurotus ostreatus cultures supplemented with copper and ferulic acid was used to decolorize the anthraquinonic dye Remazol Brilliant Blue R (RBBR). Performance of this enzymatic system was tested, and a maximum of 70% decolorization was achievable under optimal conditions. The crude preparation was immobilized by entrapment in copper alginate beads attaining 65% yield of laccase activity. Stability of the immobilized laccases was remarkably increased in comparison with that of the free enzyme preparation. Efficiency of the immobilized system was evaluated during stepwise dye additions in batch operations. Under the best conditions, 70% RBBR decolorization was achieved even after 20 cycles, although decolorization time exponentially increased after the 10th cycle. Different fixed-bed bioreactors were prepared and analyzed in continuous decolorization processes. The best performance was obtained by decreasing the amount of enzyme loaded and by improving laccase retention using chitosan-coated alginate beads.

Anthraquinones↗
PubMedDate not supplied

Modeling retrovirus production for gene therapy. 1. Determination Of optimal bioreaction mode and harvest strategy.

Although retroviruses are a promising tool for gene therapy, there are two major problems limiting the establishment of viable industrial processes: retrovirus stability and low final yield in the supernatant. This fact emphasizes the need for an effective process optimization, not only at a genetic level but also at a bioprocess engineering level. In part 1 of this paper a mathematical model was developed to optimize the bioreaction yield by determining the best retrovirus harvest strategy in perfusion cultures. PA317 cells producing recombinant retroviruses were used to develop and test this model. Cell culture was performed in stirred tanks using porous supports. The parameters of the proposed model were experimentally determined for batch and perfusion cultures at 32 and 37 degrees C both with and without additives to enhance production; the model was then validated. This model allowed the determination of the optimal values of all operational variables included: batch and perfusion duration and perfusion rate. The highest productivity (2682 virus cm(-)(3) h(-)(1)) was obtained under the following conditions: batch at 37 degrees C for 53 h followed by perfusion at 32 degrees C for 23 h with a perfusion rate of 0.107 h(-)(1). This value was 3.5-fold higher than the best result obtained in batch cultures for the same conditions of titer and quality. A sensitivity analysis of the parameters showed that the parameters that affect most the final productivity depend on the bioreaction phase: cell growth in batch culture and production and product degradation in perfusion culture. In part 2 of this paper, this model is extended to the first step of downstream processing, and the addition of further steps to the process is discussed in order to achieve global process optimization.

Animals↗
PubMed2002

Mannich-type reactions in the gas-phase: the addition of enol silanes to cyclic N-acyliminium ions.

The intrinsic gas-phase reactivity of cyclic N-acyliminium ions in Mannich-type reactions with the parent enol silane, vinyloxytrimethylsilane, has been investigated by double- and triple-stage pentaquadrupole mass spectrometric experiments. Remarkably distinct reactivities are observed for cyclic N-acyliminium ions bearing either endocyclic or exocyclic carbonyl groups. NH-Acyliminium ions with endocyclic carbonyl groups locked in s-trans forms participate in a novel tandem N-acyliminium ion reaction: the nascent adduct formed by simple addition is unstable and rearranges by intramolecular trimethylsilyl cation shift to the ring nitrogen, and an acetaldehyde enol molecule is eliminated. An NSi(CH(3))(3)-acyliminium ion is formed, and this intermediate ion reacts with a second molecule of vinyloxytrimethylsilane by simple addition to form a stable acyclic adduct. N-Acyl and N,N-diacyliminium ions with endocyclic carbonyl groups, for which the s-cis conformation is favored, react distinctively by mono polar [4(+) + 2] cycloaddition yielding stable, ressonance-stabilized cycloadducts. Product ions were isolated via mass-selection and structurally characterized by triple-stage mass spectrometric experiments. B3LYP/6-311G(d,p) calculations corroborate the proposed reaction mechanisms.

Journal Article↗