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Adverse drug reactions from birth to early childhood.

Neonates and older infants are a diverse group of children, quite different from their older counterparts. Adverse drug reactions may have profound immediate, delayed, and long-term implications for their neurologic and somatic development. The intrauterine, neonatal, and infancy periods are the only stages in life in which one is exposed to and affected by drugs administered to another person, the mother. In addition, because of the fragility of the neonate and the complexity of their illnesses, their pharmacotherapy is frequently complicated with misadventure and adverse drug reactions that are unavoidable or difficult to assess. Because of their differences in morphology and disease process and treatments, infants and children experience a different range of adverse drug reactions. These reactions are not necessarily predictable from the adult experience. Despite the advances made in the field of pediatric adverse drug reactions and the lessons learned through the misfortunes involving children, children continue to suffer. Sixty years after the Elixir of Sulfanilamide-Massengill disaster, children continue to be given medications with diethylene glycol in developing countries. Pediatricians, pharmacologists, and others must continue to be vigilant and active in preventing, monitoring, and treating adverse drug reactions in children. Learning from mistakes of the past will improve the health of children by preventing mistakes in the future.

Adult↗

Arabidopsis dihydropteroate synthase: general properties and inhibition by reaction product and sulfonamides.

Dihydropteroate synthase (DHPS) (EC 2.5.1.15) was extracted from leaves of Arabidopsis thaliana and purified 21-fold by ion-exchange chromatography. This enzyme preparation was then characterized for several of its properties. Michaelis-Menten constants for the substrates, p-aminobenzoic acid and dihydropteridine diphosphate were estimated to be 2.5 and 91 microM, respectively. In an optimized assay, the reaction product, dihydropteroic acid, competitively inhibited the enzyme activity with a Ki of 81 microM. However, neither dihydropteroic acid nor tetrahydrofolate, products further downstream in the biosynthetic pathway inhibited enzymatic activity. This appears to be the first report of product inhibition of DHPS from a higher plant. The relative inhibitory properties of several sulfonamides, analogues of p-aminobenzoic acid, were also examined. The substitutions on the amide nitrogen of the sulfonamides influenced the degree of inhibition; thus I50 values for the inhibition of the DHPS activity by sulfanilamide, sulfacetamide and sulfadiazine were estimated to be 18.6, 9.6 and 4.2 microM, respectively. The competitive pattern of inhibition was shown in experiments with sulfadiazine.

Arabidopsis↗

Iron (III) aquacomplexes as effective photocatalysts for the degradation of pesticides in homogeneous aqueous solutions.

The degradation of the herbicide asulam (4-amino-benzosulfonyl-methylcarbamate) was studied by excitation of iron (III) aquacomplexes in aqueous solutions at 365 nm as well as by exposition to solar light. Sulfanilamide was also studied as a model molecule. The initial step of asulam disappearance was shown to be due to the formation of hydroxyl radicals generated from the excitation of Fe(OH)2+, the most photoactive iron (III) monomeric species. However, when the iron (III) species was totally photoreduced to iron (II), the degradation of asulam in the presence of oxygen continued to completion. A photoreactivity of iron (II) species and/or iron (II) complexes under our experimental conditions was proposed. The experimental results indicate that the presence of iron (III), iron (II) and molecular oxygen is the condition for achieving the complete mineralization of the solution. The proposed photocatalytic cycle could provide an interesting tool for oxidations in the environment.

Carbamates↗

Carbonic anhydrase inhibitors: inhibition of human and murine mitochondrial isozymes V with anions.

In addition to sulfonamides, metal complexing anions represent the second class of inhibitors of the zinc enzyme carbonic anhydrase (CA, EC 4.2.1.1). The first inhibition study of the mitochondrial isozyme CA V (of murine and human origin) with anions is reported here. Inhibition data of the cytosolic isozymes CA I and CA II as well as the membrane-bound isozyme CA IV with a large number of anionic species such as halides, pseudohalides, bicarbonate, nitrate, hydrosulfide, arsenate, sulfamate, and sulfamidate and so on, are also provided for comparison. Isozyme V has an inhibition profile by anions completely different to those of CA I and IV, but similar to that of hCA II, which may have interesting physiological consequences. Similarly to hCA II, the mitochondrial isozymes show micro-nanomolar affinity for sulfonamides such as sulfanilamide and acetazolamide.

Animals↗

Method for construction of a simple laboratory-scale nonwoven filament biocatalytic filter

A method is described for constructing a simple laboratory-scale nonwoven filament biocatalytic filter (FBF) from multilayer acrylic vinyl acetate copolymer coated thread (yarn) potentially useful for investigation of biotransformations using entrapped enzymes or viable microorganisms. The porous structure of a commercial 128 &mgr;m, 279 denier, 100% polyester thread was sealed with a latex coat (precoat), then coated with a latex + sulfanilamide-azocasein mixture, and finally coated with a latex top coat to generate the multilayer coated thread. The FBF was constructed by drawing the thread across a 4 cm diameter rubber O-ring in the form of multiple layers and sealing the layers with silicone sealer. The FBF is combatible with a commercially available filter housing containing a flow distrtibutor. A method for characterizing the permeability of the sealant (top) coat of the FBF using azocasein release was also developed.

Journal Article↗

Construction of a thread coater and use of azocasein release To characterize the sealant coat porosity of fibers coated with latex biocatalytic coatings

A single-stage annular fiber coating method with co-current dry-air drying at 30 degrees C has been developed for multilayer coating of 128 &mgr;m diameter polyester thread (yarn) with latex films as a model for enzyme immobilization and development of a filament biocatalytic filter. Acrylic vinyl acetate polymer coatings were sequentially metered onto the fibers by the combination of a flexible squeegee and a red rubber annulus. The thread coater can operate over a range of 0.07-1.37 m/min thread velocities while delivering a nearly constant and reproducible polymer loading of 30. 8 +/- 1.3 mg/m. A 100% polyester, 278.9 denier thread was precoated with latex to generate an approximately 369 denier sealed filament. The filament was then coated with a latex + sulfanilamide-azocasein mixture and sealed with a polymer top coat. The permeability of the polymer sealant top coat was characterized using entrapped azocasein as a tracer molecule and monitoring the azocasein release upon rehydration of the coated threads. Azocasein release rate decreased with curing time at 30 degrees C until 2 days and was invariant after 2-3 days of curing. A 282 mOsm rehydrating solution was sufficient to suppress increased azocasein release due to top coat blistering. No enhancement in the permeability of the top coat was observed when high molecular weight water soluble polymers (WSPs) were used as fillers. This probably results from the low WSP to latex ratio used (0.05-0.1) and the slow rate of WSP leaching compared to the release of azocasein. A method using 60-120 mesh silica was also developed to study the effect of mechanical abrasion of the coated threads as measured by azocasein release kinetics.

Journal Article↗

Preserving the chromatographic integrity of high-speed supercritical fluid chromatography separations using time-of-flight mass spectrometry.

The advantage of high-speed time-of-flight-mass spectrometry (TOF-MS) detection for ultrafast qualitative supercritical fluid chromatography/mass spectrometry (SFC/MS) applications allows the superior resolving power of SFC to be exploited in high-throughput analysis. A chromatographic comparison of quadrupole MS and TOF-MS shows high-speed TOF total ion current data point sampling to be more indicative of fast SFC separations and corresponding short (1-2 s) baseline peak widths. Results shown for analysis of a six-compound mixture with two peaks eluting at 0.86 and 0.89 min exhibit >50% resolution by high-speed TOF data sampling, whereas the same peaks appear to coelute using quadrupole MS data sampling. Additionally, a marked improvement in the peak baseline widths is afforded by fast TOF data acquisition of 0.1 s/spectrum, resulting in a reduction in the baseline width, 1.6 s, of sulfanilamide in a four-compound mixture that is more than 2-fold greater than that achieved at the slower data acquisition of 0.5 s/spectrum. The resulting increase in resolution and improved peak shapes allow automatic integration routines to perform more effectively. For most classes of compounds amenable to high performance liquid chromatography, including druglike species, steroids, and polymers, the union of SFC with TOF-MS provides the maximum density of chemical information per unit time available with any high-speed chromatographic/mass spectrometric method.

Journal Article↗

Generation of class-selective monoclonal antibodies against the penicillin group.

To develop a penicillin-specific ELISA, different attempts were made to obtain monoclonal antibodies specific for the common structure of penicillins. Ampicillin was coupled to different carrier proteins (bovine serum albumin, chicken ovalbumin, and thyroglobulin) to render it immunogenic. Different coupling methods were compared: two methods using a cross-linker (glutaraldehyde or a succinimide ester), one carbodiimide-mediated coupling method, and one method without any cross-linker or mediator molecule (physiological binding). Mice were immunized with the conjugates intraperitoneally or in the footpad. A screening ELISA was developed to detect anti-ampicillin antibodies in sera. Specificity and affinity of the antibodies were demonstrated by inhibiting their binding with a 10 mM solution of ampicillin. No difference could be observed using electrofusion or PEG-mediated fusion. For the production of the monoclonals, an intravenous final boost gave antibodies with better specificity and affinity than an intraperitoneal final booster injection. At least one anti-ampicillin monoclonal antibody (19C9) cross-reacts with penicillin G, oxacillin, dicloxacillin, and carbenicillin, and not with sulfanilamide, chloramphenicol, neomycin, and streptomycin, and is therefore considered interesting for developing a penicillin-specific ELISA.

Animals↗

Structure-activity relationship of glycine betaine analogs on osmotolerance of enteric bacteria.

Bacterial cells have the ability to accumulate compatible solutes within the cytoplasm to maintain their osmolarity above that of the extracellular milieu. Glycine betaine (GB) and its biosynthetic precursor choline (Chol) are the major compatible solutes that bacteria accumulate when osmotically challenged. Different osmotically triggered active transport mechanisms have been identified for GB and Chol. In the present study we examined the bioisosteric replacement of the carboxylic group of GB with sulfonic, phosphonic or benzenesulfonamido groups. The sulfonic acid analog (sulfobetaine, compound 3) showed osmoprotectant activity equivalent to that of GB. In addition, we tested the possibility of utilizing GB/Chol transport systems to deliver cytotoxic analogs of GB into three strains of E. coli that differed in their salt resistance. We found that N1-betainyl-N4-(haloacetyl)sulfanilamides (compounds 17c-e) that are GB analogs containing alkylating side chain within their structures inhibited the bacterial growth of the tested standard and salt sensitive strains of E. coli. We also showed that the (N-methyl-cyclic ammonio)methanesulfonates (compounds 21a-c) are able to block Chol transport system in both the standard and the salt-sensitive E. coli strains used. At the concentration used (0.1 mM), none of the tested compounds showed any significant effect on the salt-resistant strain used.

Betaine↗

Carbonic anhydrase inhibitors. The mitochondrial isozyme VB as a new target for sulfonamide and sulfamate inhibitors.

A lately discovered carbonic anhydrase (hCA, EC 4.2.1.1), the mitochondrial hCA VB, was cloned, expressed, and purified. Kinetic parameters proved it to be 3.37 times more effective than hCA VA as a catalyst for the physiological reaction, with kcat = 9.5 x 10(5) s(-1) and kcat/K(M) = 9.8 x 10(7) M(-1) s(-1), being second only to hCA II among the 16 isoforms presently known in humans. We investigated the inhibition of hCA VB with a library of sulfonamides/sulfamates, some of which are clinically used compounds. Benzenesulfonamides were ineffective inhibitors, whereas derivatives bearing 4-amino, 4-hydrazino, 4-methyl, 4-carboxy moieties or halogenated sulfanilamides were more effective (Ki's of 1.56-4.3 microM). Among the 10 clinically used compounds, acetazolamide, benzolamide, topiramate, and indisulam showed effective inhibitory activity (Ki's of 18-62 nM). Three compounds showed better activity against hCA VB over hCA II, among which were sulpiride and ethoxzolamide, which were 2 times more effective inhibitors of the mitochondrial over the cytosolic isozyme. hCA VB is a druggable target and some of its inhibitors may lead to the development of novel antiobesity therapies.

Amino Acid Sequence↗

Carbonic anhydrase inhibitors: DNA cloning and inhibition studies of the alpha-carbonic anhydrase from Helicobacter pylori, a new target for developing sulfonamide and sulfamate gastric drugs.

We have cloned and sequenced Helicobacter pylori alpha-class carbonic anhydrase (hpCA) from patients with different gastric mucosal lesions, including gastritis (n=15), ulcer (n=6), and cancer (n=16). Although several polymorphisms were newly identified such as 12Ala, 13Thr, 16Ile, and 168Phe, there was no significant relevance of any polymorphism with gastric mucosal lesion types. A library of sulfonamides/sulfamates has been investigated for the inhibition of hpCA, whereas new derivatives have been obtained by attaching 4-tert-butyl-phenylcarboxamido/sulfonamido tails to benzenesulfonamide/1,3,4-thiadiazole-2-sulfonamide scaffolds. All types of activity for inhibition of hpCA have been detected. Dorzolamide and simple 4-substituted benzenesulfonamides were weak inhibitors (KI 873-4360 nM). Sulfanilamide, orthanilamide, some of their derivatives, and indisulam showed better activity (KI 413-640 nM), whereas most of the clinically used inhibitors, such as methazolamide, ethoxzolamide, dichlorophenamide, brinzolamide, topiramate, zonisamide, etc., acted as medium-potency inhibitors (KI 105-378 nM). Some potent hpCA inhibitors were detected too (KI 12-84 nM) among acetazolamide, 4-amino-6-chloro-1,3-benzenedisulfonamide and some newly designed compounds incorporating lipophilic tails. Some of the newly prepared derivatives had selectivity ratios for inhibiting hpCA over hCA II in the range of 1.25-3.48, showing thus some selectivity for inhibiting the bacterial enzyme. Since hpCA is essential for the survival of the pathogen in acid, it might be used as a new pharmacologic tool in the management of drug-resistant H. pylori.

Amino Acid Sequence↗

Carbonic anhydrase inhibitors. DNA cloning, characterization, and inhibition studies of the human secretory isoform VI, a new target for sulfonamide and sulfamate inhibitors.

The secretory isozyme of human carbonic anhydrase (hCA, EC 4.2.1.1), hCA VI, has been cloned, expressed, and purified in a bacterial expression system. The kinetic parameters for the CO2 hydration reaction proved hCA VI to possess a kcat of 3.4 x 10(5) s-1 and kcat/KM of 4.9 x 10(7) M-1 s-1 (at pH 7.5 and 20 degrees C). hCA VI has a significant catalytic activity for the physiological reaction on the same order of magnitude as the ubiquitous isoform CA I or the transmembrane, tumor-associated isozyme CA IX. A series of sulfonamides and one sulfamate have been tested for their interaction with this isozyme. Simple benzenesulfonamides were rather ineffective hCA VI inhibitors, with inhibition constants in the range of 1090-6680 nM. Better inhibitors were detected among such derivatives bearing 2- or 4-amino-, 4-aminomethyl-, or 4-hydroxymethyl moieties or among halogenated sulfanilamides (KI values of 608-955 nM). Some clinically used compounds, such as acetazolamide, methazolamide, ethoxzolamide, dichlorophenamide, dorzolamide, brinzolamide, topiramate, sulpiride, and indisulam, or the orphan drug benzolamide, showed effective hCA VI inhibitory activity, with inhibition constants of 0.8-79 nM. The best inhibitors were brinzolamide and sulpiride (KI values of 0.8-0.9 nM), the latter compound being also a CA VI-selective inhibitor. The metallic taste reported as a side effect after the treatment with systemic sulfonamides may be due to the inhibition of the salivary CA VI. Some of the compounds investigated in this study might be used as additives in toothpastes for reducing the acidification produced by the relevant CO2 hydrase activity of enamel CA VI, which leads to the formation of protons and bicarbonate and may have a role in cariogenesis.

Amino Acid Sequence↗

Magnetic Field Orientation of Liquid Crystalline Epoxy Thermosets.

The effect of magnetic fields on the orientation and properties of 4,4'-bis(2,3-epoxypropoxy)-alpha-methylstilbene cured with sulfanilamide has been studied. This epoxy system is initially isotropic and forms a smectic A phase upon curing. A magnetic field was applied during the cure reaction, resulting in alignment of the molecules along the direction of the applied field. Measurement of the orientation parameter of the fully cured material by wide-angle X-ray scattering (WAXS) showed that orientation improved with an increase in field strength. The orientation parameters of the smectic layer normals calculated from the inner reflection of the WAXS pattern attained a maximum level of approximately 0.8 at a field strength of approximately 12 T. The orientation parameters calculated from the outer reflection of the WAXS pattern were considerably lower, possibly due to the presence of amorphous regions associated with domain boundaries or the loss of molecular alignment within the smectic layers due to topological restrictions of the cross-linking sites. Orientation resulted in an anisotropic linear thermal expansion coefficient after curing, although the overall volumetric expansion was constant. The elastic tensile modulus increased with the square of the orientation parameter, attaining a maximum value of 8.1 GPa, compared to 3.1 GPa for the unoriented material. The change in modulus with orientation could be fit with a simple model for the modulus of anisotropic materials.

Journal Article↗

Flow-through photometric sensor for determination of sulfonamides.

A sensor based on integration of spectrophotometric detection and retention of the product of the Bratton-Marshall reaction for the determination of sulfanilamide, sulfamethazine, sulfaquinoxaline and sulfathiazole is reported. The methods developed using this sensor in a flow injection manifold feature linear ranges for these analytes between 10 and 1200 ng ml(-1) with relative standard deviations smaller than 2.6% in all instances and they were applied to the determination of sulfonamides in spiked milk and water samples with excellent results. The two main advantages of these sensor-based methods over other automated and non-automated counterparts are enhanced sensitivity as a result of in situ concentration and avoidance of using a de-bubbler to eliminate the nitrogen produced in the flowing system.

Animals↗

WY-50295 tromethamine: a 5-lipoxygenase inhibitor without activity in human whole blood.

The 5-LO inhibitor, WY-50295 tromethamine (T) prevented leukotriene release (LTB4 production) in calcium ionophore stimulated, purified human and rat neutrophils. However, whereas WY-50295T inhibited both in vitro and ex vivo rat whole blood leukocyte LTB4 formation (IC50= 40 microM and oral ED50 of 18 mg/kg, respectively), it did not inhibit LTB4 production in calcium ionophore stimulated human whole blood at concentrations to 200 microM. To reduce binding of WY-50295T to serum albumin, 250 microM of a naphthalene sulfonic acid (> 99.9% binding to albumin primarily at the carboxylic site) and 250 microM sulfanilamide (binding to nonspecific sites) separately or in combination were preincubated in whole blood prior to addition of WY-50295T; however, WY-50295T still did not inhibit 5-LO and free drug blood levels were unchanged. When purified human neutrophils in the presence of fatty acid saturated albumin (fraction V) was employed, the 5-LO inhibitory activity of WY-50295T was prevented. Zileuton (5 microM) inhibited LTB4 production by 99% in the presence of these albumins. Also, rat albumin presented WY-50295T to purified rat neutrophils more effectively than human albumin (i.e. WY-50295T was more active in the presence of rat albumin). These results suggest that the high affinity binding of WY-50295T to human albumin and possibly the reduction of drug uptake (passive diffusion) using purified human vs rat neutrophils may account for the inactivity of WY-50295T in the human whole blood assay.

Animals↗

Structure of hemoglobins Zürich [His E7(63)beta replaced by Arg] and Sydney [Val E11(67)beta replaced by Ala] and role of the distal residues in ligand binding.

In hemoglobin Zürich the side chain of the distal arginine attaches itself to the propionate of the heme, leaving the heme pocket wide open, allowing sulfanilamides easy access to the iron, and doubling the partition coefficient between CO and O2. The replacement of the distal valine by alanine in hemoglobin Sydney leaves a large gap inside the heme pocket, which is partly filled by a water molecule bonded to the distal histidine. Hemoglobin Sydney has the same partition coefficient between CO and O2 as hemoglobin A. Replacement of the distal histidine increases the stretching frequency of CO linked to the beta heme by 6 cm-1, but replacement of the distal valine increases it by only 3 cm-1, but replacement of the distal histidine leaves the O-O stretching frequency unchanged.

Alanine↗

A modified procedure for the determination of nitrate in sediments and some natural waters.

This paper describes a modified procedure for the determination of nitrate in sediments and some natural water samples. In the Water Quality Branch Laboratories where large numbers of samples are regularly analysed, it was found that existing methods1,4 do not give precise and accurate results particularly when applied to sediment samples and some natural waters. The reasons for such inaccuracies have sebsequently been found and a method for removal of such interfering substances have been devised. The proposed method essentially consist of two steps. In the first step the interfering substances such as sulfide and humic acid substances are removed by precipitation with copper. The resultant solution is then analysed by a colorimetric approach utilizing cadmium fillings as the reducing agent to reduce nitrate to nitrite, and analyse the resultant nitrite with sulfanilamide, and naphtylene diamine. The smallest amount of nitrate that can be detected with certainty is approximately 10 mug-at N/Kg and 50 mug/ at N/Kg in water and sediments respectively. The figures for sediments are based on dissolving 20 gms. of sediment, dry weight, in 100 ml volume. The proposed method has been tested on a wide variety of natural water samples, sediments, clays and sands.

Chemical Precipitation↗

Low-temperature sealed tube combustion of gaseous, liquid and solid organic compounds for 13C/12C and 14C analysis.

A new, low-temperature sealed tube technique for combustion of organic carbon prior to subsequent off-line isotope analysis is proposed. Complete oxidation is achieved with potassium peroxodisulfate and silver permanganate as oxidants at temperatures not exceeding 500 degrees C. The combustion of gaseous (methane), solid (cane sugar, vanilla, N-thiazolyl-2-sulfamide, ascorbic acid, phenanthrene, thiourea, polyethylenefilm, tetrafluoropolyethylene, polyetheretherketone, graphite, and Suwannee River Fulvic Acid), and liquid (tetrachloroethene, toluene, and oil) model compounds and international standards was tested. A 24 h combustion at 500 degrees C was sufficient for complete oxidation in all cases. The time required for complete oxidation of Suwannee River Fulvic Acid, typical of refractory freshwater dissolved organic carbon, as a function of combustion temperature was 2 h at 500 degrees C, 6 h at 400 degrees C, and 24 h at 300 degrees C. Preparation of saline solution parallels of cane sugar, vanilla, N-thiazolyl-2-sulfanilamide, and ascorbic acid gave consistent results. For reproducible delta13C analyses using a Thermoquest MAT 252 MS, a minimum of 5 microg C had to be combusted. Reliable 14C results, measured at an accelerator mass spectrometer facility, were obtained from coal and from cane sugar combusted for 24 h at 500 degrees C by the proposed method.

Carbon↗