Abstract  It is still a challenge to link specific metabolic activities to certain species in a microbial community because of methodological limitations. We developed a method to analyze the specific metabolic activity of a single bacterial species within a consortium making use of [(13)C(7)]-toluene for metabolic labelling of proteins. Labelled proteins were subsequently analyzed by 2D gel electrophoresis (2-DE) and mass spectrometry (MS) to characterize their identity as well as their (13)C content as an indicator for function and activity of the host organism. To establish this method, we analyzed the metabolic incorporation of (13)C carbon atoms into proteins of Aromatoleum aromaticum strain EbN1. This strain is capable of metabolizing toluene under nitrate-reducing conditions and was grown in either pure culture or in a mixed consortium with a gluconate-consuming enrichment culture. First, strain EbN1 was grown with non-labelled toluene or labelled [(13)C(7)]-toluene as carbon sources, respectively, and their proteins were subjected to 2-DE. In total, 60 unique proteins were identified by MALDI-MS/MS. From 38 proteins, the levels of (13)C incorporation were determined as 92.3+/-0.8%. Subsequently, we mixed strain EbN1 and the enrichment culture UFZ-1, which does not grow on toluene but on gluconate, and added non-labelled toluene, [(13)C(7)]-toluene and/or non-labelled gluconate as carbon sources. The isotope labelling of proteins was analyzed after 2-DE by MS as a quantitative indicator for metabolic transformation of isotopic-labelled toluene by the active species of the consortium. Incorporation of (13)C was exclusively found in proteins from strain EbN1 at a content of 82.6+/-2.3%, as an average calculated from 19 proteins, demonstrating the suitability of the method used to identify metabolic active species with specific properties within a mixed culture.

Abstract  Ion channels from the midgut apical membrane of gypsy moth (Lymantria dispar) larvae were studied following mechanical fusion of brush-border membrane vesicles with planar phospholipid bilayer membranes. In symmetrical 300 mmol l(-)(1) KCl (pH 9.0), nine different channels with conductances ranging from 27 to 795 pS and linear current/voltage relationships were resolved. In the presence of a KCl gradient across the bilayer (450 mmol l(-)(1 )cis/150 mmol l(-)(1 )trans), 11 different conductance levels ranging from 16 to 850 pS were detected. The channels were slightly cationic: the zero-current reversal potential was shifted by -5 mV to -21 mV compared with symmetrical KCl conditions, corresponding to p(K)/p(Cl) permeability ratios of 1.5-8.0. Most channels were neither voltage-dependent nor Ca(2+)-sensitive and displayed complex gating kinetics. Addition of Ba(2+) or Cs(+) to both sides of the bilayer had little effect on channel activity, but fewer distinct channels were observed when KCl was replaced by potassium gluconate, suggesting an effect of Cl(-) on channel activity. A reduced number of channels was also detected when KCl was replaced by N-methyl- d-glucamine-HCl. Under asymmetrical N-methyl-d-glucamine-HCl conditions, only anionic channels were observed. They exhibited current rectification (35 pS at negative voltages and 81 pS at positive voltages) and were strongly voltage-dependent.

Abstract  The Na(+)/H(+) exchanger has been the only unequivocally demonstrated H(+)-transport mechanism in the synaptosomal preparation. We had previously suggested that a Cl(-)-H(+) symporter (in its acidifying mode) is involved in cytosolic pH regulation in the synaptosomal preparation. Supporting this suggestion, we now show that: (1) when synaptosomes are transferred from PSS to either gluconate or sulfate solutions, the Fura-2 ratio remains stable instead of increasing as it does in 50 mM K solution. This indicates that these anions do not promote a plasma membrane depolarization. (2) Based in the recovery rate from the cytosolic alkalinization, the anionic selectivity of the Cl(-)-H(+) symporter is NO(3)(-) > Br(-) > Cl(-) > I(-) = isethionate = sulfate = methanesulfonate = gluconate. (3) PCMB 10 muM inhibits the gluconate-dependent alkalinization by 30 +/- 6%. (4) Neither Niflumic acid, 9AC, Bumetanide nor CCCP inhibits the recovery from the cytosolic alkalinization.

Abstract  Pseudomonas putida morphine dehydrogenase is shown to be closely homologous to 18 proteins, defining a superfamily within which morphine dehydrogenase particularly resembles two bacterial, 2,5-dioxo-D-gluconic acid reductases, and two eukaryotic proteins of unknown functions. Relationships within the superfamily are extensive and complex. Residue identities between protein pairs range from 29-90%. Three subgroups are proposed. Nevertheless, on the basis of residue conservations/exchanges it is suggested that the nicotinamide coenzyme binding and substrate reduction occur in all the enzymes by broadly analogous mechanisms, among which some probable differences are identified.

Abstract  The aim of this study was to investigate the role of process conditions and system composition on the acid-induced gelation of a mixture of milk protein and gum tragacanth. This was studied by determining the effects of co-solute (lactose) addition (3, 5 and 7%) and gelation temperature (25, 37 and 45°C) on the mixture's rheological properties and microstructure using a combination of techniques including small-deformation rheology and scanning electron microscopy. The presence of lactose played an important role in the microstructure formation of gels but did not change most rheological properties. The microstructure of gels formed in the presence of lactose was coarser and more particulate, but less interconnected; this can be explained by lactose's role in improving protein aggregation. Gels prepared at a lower temperature had a high structure strength, as indicated by their high storage modulus, τ(f) and G(f) values. Low gelation temperature also caused a more branched and homogenous microstructure.

Abstract  Butyrate has an antitumorigenic effect on colorectal cancer cell lines. Dietary sodium gluconate (GNA) promotes butyrate production in the large intestine. Accordingly, we examined the effect of dietary GNA on tumorigenesis in the large intestine in rats. Male Fisher-344 rats (n = 32) were divided into 4 groups: 2 diets (with or without 50 g GNA/kg basal diet) X 2 treatments (with or without carcinogen administration). Colonic tumors were induced by 3 intraperitoneal injections of azoxymethane (115 mg/kg body wt, 1 time/wk) and dietary deoxycholic acid (2 g/kg basal diet). The experiment was conducted for 33 wk except for a few rats. Ingestion of GNA increased cecal butyrate concentration at the end of experiment (P < 0.01). No tumor development occurred in the untreated groups. Ingestion of GNA decreased the incidence of tumors in rats administered the carcinogen (37.5 vs. 100%, P < 0.05). Ingestion of GNA also decreased the mean number of tumors per rat (0.5 +/- 0.8 vs. 2.8 +/- 1.5, P < 0.01). beta-Catenin accumulation and TdT-mediated dUTP nick end labeling (TUNEL) positive cells in tumors were histochemically examined. The results of this study suggested that the antitumorigenic effect of GNA may involve the stimulation of apoptosis through enhanced butyrate production in the large intestine.

Abstract  Biomasses of methylotrophic bacteria, yielded by biotechnological processes as waste products, can represent a source of ubiquinones, especially of the ubiquinone-10. Possibilities for the separation of ubiquinones were studied on waste biomasses from the microbial production of gluconic acid, from the production of intracellular poly-beta-hydroxybutyric acid (PHB) and from the sewage treatment. The ubiquinones are extracted with supercritical CO2 in the presence of methanol or ethanol as entrainer. The separation of ubiquinones with supercritical extraction was more effective than with conventional extraction methods. The advantages are especially the low extract quantities and the high ubiquinone content in the extracts. The crude ubiquinones can be purified by using the preparative HPLC technique.

Abstract  We assessed mechanisms of acetylcholine- and bradykinin-induced relaxations in human omental resistance vessels. Ring segments (approximately 200 microns normalized ID) were dissected from omental biopsies obtained from women at laparotomy (nonpregnant) or at cesarean delivery (pregnant) and were studied under isometric conditions in a Mulvany-Halpern myograph. All arginine vasopressin-preconstricted vessels relaxed in a strictly endothelium-dependent manner to acetylcholine and bradykinin; maximal relaxations were not decreased by either NG-nitro-L-arginine or indomethacin. By contrast, bradykinin failed to relax vessels that had been preconstricted with potassium gluconate. In the combined presence of NG-nitro-L-arginine and indomethacin, addition of charybdotoxin, a selective antagonist of some calcium-sensitive potassium channels, did not inhibit maximal bradykinin-induced relaxation. By contrast, addition of 10 mmol/L tetraethylammonium chloride abolished relaxation in vessels from nonpregnant women but not in vessels from gravidas. We conclude that bradykinin relaxes these human resistance arteries in an endothelium-dependent but predominantly nitric oxide- and prostanoid-independent manner; relaxation likely depends on the action of an endothelium-derived hyperpolarizing vasodilator. Furthermore, in striking contrast to mechanistic insights from animal studies, human pregnancy appears to augment a mechanism of endothelium-dependent relaxation in these vessels that is insensitive to the inhibitors noted above. Whether a similar novel vasodilator mechanism in vivo contributes to the physiological vasodilation that characterizes human gestation or whether failure of such a mechanism might lead to preeclampsia remains the subject of future study.

Abstract  Membrane-integrated quinoprotein glucose dehydrogenase from Acinetobacter calcoaceticus was produced by heterologous expression of the gene for it in an Escherichia coli recombinant strain. The apoenzyme (lacking the cofactor pyrroloquinoline quinone, PQQ) was solubilized with Triton X-100 and purified to homogeneity. Reconstitution of the apoenzyme to full activity in the assay was achieved with a stoichiometric amount of PQQ in the presence of Mg2+. Just as for other PQQ-containing dehydrogenases where Ca2+ fulfills this role, Mg2+ anchors PQQ to the mGDH protein and activates the bound cofactor. This occurs in a precise way since high anomer specificity was found for the enzyme toward the sugars tested. Although the steady-state-type kinetics were as expected for a dye-linked dehydrogenase (ping-pong) and the PQQ in it was present in oxidized form, addition of glucose to the holoenzyme resulted in a very slow but continuous production of gluconolactone; i.e., the reaction did not stop after one turnover, with O2 apparently acting as an (albeit poor) electron acceptor by reoxidizing PQQH2 in the enzyme. The surprisingly low reactivity with glucose, in the absence of dye, as compared to the activity observed in the steady-state assay appeared to be due to formation of an anomalous enzyme form, mGDH. Formation of normal holoenzyme, mGDH, reducing added glucose immediately to gluconolactone (in one turnover), was achieved by treating mGDH with sulfite, by reconstituting apoenzyme with PQQ in the presence of sulfite, or by applying assay conditions to mGDH (addition of PMS/DCPIP). As compared to other quinoprotein dehydrogenases, mGDH appears to be unique with respect to the mode of PQQ-binding, as expressed by the special conditions for reconstitution and the absorption spectra of the bound cofactor, and the reactivity of the reduced enzyme toward O2. The primary cause for this seems not to be related to a different preference for the activating bivalent metal ion but to the special way of binding of PQQ to mGDH.

Abstract  PESTAB Therapy of chronic poisonings with organochlorine compounds, such as DDT, BHC, 2-KF, dichloralurea, 2,4-D, polychloropinene, and hexachlorobutadiene, is reviewed. Since the mechanism of action of organochlorine compounds in humans is not known, specific antidote therapy is not available. Glucose, calcium gluconate, and vitamins, especially vitamin B, are useful during the initial stage of the chronic poisoning, characterized by autonomic dystonia and the asthenic syndrome. Belladonna preparations are used for elimination of autonomic irritability and of vasculay spasms, and tranquilizers are given to reduce emotional lability. Vitamin preparations, cocarboxylase, adenosine triphosphatase, and oxygen therapy are used to control hypoxia and to increase tonus. Electrophoresis according to Shcherbak with calcium chloride, vitamin B1, dimedrol, or novocaine is also useful. Vitamin B12 is given to control toxic polyneuritis. Hemorrhagic vasculitis is treated with vitamins C and P. B vitamins, thiamine, riboflavin, glucose, and insulin are used for the therapy of toxic hepatitis. Impairments of gastric secretory function are corrected by specific diets, nicotinic acid, and pyridoxine. Vitamins C and B, thiamine, folic acid, and iron preparations are useful in anemia.

Abstract  The production of citric and gluconic acids from fig by Aspergillus niger ATCC 10577 in solid-state fermentation was investigated. The maximal citric and gluconic acids concentration (64 and 490 g/kg dry figs, respectively), citric acid yield (8%), and gluconic acid yield (63%) were obtained at a moisture level of 75%, initial pH 7.0, temperature 30 degrees C, and fermentation time in 15 days. However, the highest biomass dry weight (40 g/kg wet substrate) and sugar utilization (90%) were obtained in cultures grown at 35 degrees C. The addition of 6% (w/w) methanol into substrate increased the concentration of citric and gluconic acid from 64 and 490 to 96 and 685 g/kg dry fig, respectively.

Abstract  1. Although calcium gluconate (CG) is recommended in the treatment of hydrofluoric acid (HF) eye burn its efficacy seems to be controversial, and controlled human or animal studies are limited. The study's objective is to compare the efficacy of 1% CG and normal saline irrigation for the treatment of HF eye injury in animals. 2. 0.05 ml 2% HF was instilled to anesthetized rabbit's eyes. One minute later, four treatment groups were studies: (1) irrigation with normal saline followed by topical antibiotics, corticosteroids and cycloplegics for 48 h (n = 10); (2) irrigation with 1% CG followed by the same topical treatment (n = 9); (3) as group 1 and 1% CG drops over 48 h (n = 10); (4) as group 3, and injection of 1% CG subconjunctivally after irrigation (n = 9). 3. Corneal erosion area, corneal haziness, conjunctival status, vascularization (pannus) and acidity were assessed before injury, immediately after initial treatment and 1, 2, 7 and 14 days thereafter by slit lamp aided by fluorescein staining. 4. Conjunctival pH dropped from 6.0-6.5 to 2.5-3 after injury and increased to 6-6.5 after irrigation. Corneal erosion: smaller in groups 2, 3, significantly so at 2 days, but not different at 14 days. Corneal haziness: more severe in group 4, at 14 days, insignificant. Conjunctival damage: significantly worse in group 4 at 2, 7 and 14 days. Pannus appeared in 2-4 eyes in each group. 5. It seems that for HF injury 1% CG did not have any significant advantage over saline irrigation and topical treatment only. It might have some initial and temporary effect on healing process especially that involving erosion. Given subconjunctivally, 1% CG may be toxic and worsens clinical outcome.

Abstract  The regulation of PTH secretion by calcium is altered in patients with primary hyperparathyroidism. A similar disturbance may occur in secondary hyperparathyroidism, but direct in vivo comparisons of PTH secretion in normal subjects and those with secondary hyperparathyroidism have not been made. Thus, 13 patients with end-stage renal failure and secondary hyperparathyroidism and 20 healthy volunteers underwent dynamic tests of PTH secretion. Changes in ionized calcium were induced by 2-h iv infusions of calcium gluconate or sodium citrate on consecutive days, and the sigmoidal relationship between serum ionized calcium and PTH levels was examined. During sodium citrate infusions, serum ionized calcium levels decreased by 0.21 +/- 0.04 and 0.20 +/- 0.05 mmol/L, respectively (mean +/- SD), in normal volunteers and dialyzed patients (P = NS). Serum PTH levels rose from 27 +/- 7 to 107 +/- 33 pg/mL in controls and from 480 +/- 238 to 859 +/- 412 pg/mL in dialyzed subjects; thus, maximum PTH levels were 396% of preinfusion values in normal subjects, but only 79% greater than baseline values in dialyzed patients (P < 0.001). During the first 30 min of calcium infusions, the increase in serum ionized calcium did not differ between groups, but PTH levels fell more rapidly in normal volunteers; values were 24% of preinfusion levels in controls, but only 56% of the baseline in dialyzed patients (P < 0.01) after 30 min. Minimum PTH levels were attained after 50 min of calcium infusion in normal volunteers and after 70 min in dialyzed patients. The derived values for set-point were 1.21 +/- 0.04 and 1.24 +/- 0.06 mmol/L, respectively, in control and dialyzed subjects (P = NS). These results do not support the contention that the set-point for calcium-regulated PTH secretion is greater than normal in patients with secondary hyperparathyroidism due to end-stage renal disease.

Abstract  BACKGROUND: Patients' exposure to and potential toxicity from aluminum in parenteral nutrition (PN) formulations is an important concern of healthcare providers.

OBJECTIVE: To determine the potential for aluminum toxicity caused by PN in hospitalized adults who have risk factors of both acute kidney injury and PN.

METHODS: Adults who required PN and had a serum creatinine (SCr) level at least 1.5 times greater than the admission SCr on the first day of PN were studied in a retrospective fashion. Protein was administered based on whether hemodialysis was being used (0.6-1 g/kg/day without hemodialysis; 1.2-1.5 g/kg/day with hemodialysis). Aluminum exposure was determined for each patient by multiplying the volume of each PN component by its concentration of aluminum. Unpaired t-tests, Fisher's exact test, and analysis of variance were used for statistical analysis. Data are presented as mean +/- SD.

RESULTS: Thirty-six patients (aged 50.4 +/- 20.4 y; weight 90.2 +/- 32.8 kg) were studied. Initial serum urea nitrogen and SCr were 47 +/- 23 and 3.3 +/- 1.4 mg/dL, respectively. Twelve patients received hemodialysis. The mean aluminum exposure was 3.8 +/- 2 microg/kg/day in the 36 patients. Of these, 29 had safe calculated aluminum exposure (<5 microg/kg/day) and 7 had high calculated aluminum exposure (>5 microg/kg/day). Patients with safe aluminum exposure had significantly higher SCr levels than did those with high aluminum exposure (3.5 +/- 1.5 vs 2.2 +/- 0.7 mg/dL; p < 0.04). Patients with high aluminum exposure received significantly more aluminum from calcium gluconate compared with those who had safe aluminum exposure (357 +/- 182 vs 250 +/- 56 microg/day; p < 0.02). Limitations of the study include its retrospective design, which resulted in calculated versus direct measurement of aluminum.

CONCLUSIONS: Using our calculations, we believe that most patients with acute kidney injury who require PN do not receive excessive exposure to aluminum from the PN formulation, despite having 2 risk factors (acute kidney injury, PN) for aluminum toxicity.

Abstract  In this study, an attempt was made to identify an effective phosphate solubilizing bacteria from pesticide polluted field soil. Based on the formation of solubilization halo on Pikovskaya's agar, six isolates were selected and screened for pesticide tolerance and phosphate (P) solubilization ability through liquid assay. The results showed that only one strain (SGRAJ09) obtained from Achillea clavennae was found to tolerate maximum level of the pesticides tested and it was phylogenetically identified as Pseudomonas sp. It possessed a wide range of pesticide tolerance, ranging from 117 μg mL(-1) for alphamethrin to 2,600 μg mL(-1) for endosulfan. The available P concentrations increased with the maximum and double the maximum dose of monocrotophos and imidacloprid, respectively. On subjected to FT-IR and HPLC analysis, the presence of organic acids functional group in the culture broth and the production of gluconic acid as dominant acid aiding the P solubilization were identified. On comparison with control broth, monocrotophos and imidacloprid added culture broth showed quantitatively high organic acids production. In addition to gluconic acid production, citric and acetic acids were also observed in the pesticide amended broth. Furthermore, the Pseudomonas sp. strain SGRAJ09 possessed all the plant growth promoting traits tested. In presence of monocrotophos and imidacloprid, its plant growth promoting activities were lower than that of the pesticides unamended treatment.

Abstract  Endogenous enteric nitric oxide has multiple functions. Enteric nitric oxide may be diminished in the premature infant and may therefore predispose the immature intestine to injury. The aim of this study was to determine if the infusion of a nitric oxide donor (nitroglycerin) would attenuate intestinal damage in a rabbit model of necrotizing enterocolitis. Transmural injection of rabbit intestinal loops with an acidified solution of casein and calcium gluconate simulates certain aspects of necrotizing enterocolitis. After injection of acidified casein solution into rabbit intestinal loops, twelve rabbits were randomly divided into two groups: six received maintenance fluids only and six received maintenance fluids and a nitroglycerin infusion adjusted to maintain mean arterial pressure 10 mm Hg below baseline (range, 2 to 12 micrograms/kg/min). After 3 hours, the rabbits were killed, and the intestinal tissue graded histologically. Intestinal damage in the nitroglycerin-treated rabbits was significantly less than that of untreated controls (mean histological grade of 0.39 v 1.48, P < .001). In this rabbit model of necrotizing enterocolitis, infusion of the nitric oxide donor nitroglycerin significantly attenuates intestinal damage. We speculate that enteric nitric oxide deficiency, as may exist in the preterm infant, predisposes the intestine to necrotizing enterocolitis.

Abstract  A simplified kiet has been developed for 99mTc protein radiolabeling using an N3S triamide mercaptide bifunctional chelating agent and the preformed chelate approach. The process combined N3S chelating agent, gluconate intermediate transfer agent, stannous reducing agent, and gentisic acid stabilizer into a lyophilized formulation. With sulfur donor atom hemithioacetal protection of the ligand, delta-2,3,5,6-tetrafluorothiophenyl alpha-S-(1-ethoxyethyl)mercaptoacetamido-L-adipoylglycylglycine , optimum 99mTc chelation was achieved in a single step. Subsequent reaction with NR-LU-10 antibody Fab fragment followed by purification via QAE Sephadex anion exchange resin filter afforded 99mTc-N3S-NR-LU-10 Fab conjugate with retained immunoreactivity and effective tumor targeting properties.

Abstract  The mechanism responsible for the transport of the glutathione-methylmercury complex (CH3HgSG) across liver canalicular membranes into bile was investigated. Uptake of radiolabeled CH3HgSG by canalicular liver plasma membranes (cLPM) isolated from Sprague-Dawley-rats was dependent upon time and temperature. To reduce nonspecific binding of the complex by the membrane, S-(2,4-dinitrophenyl)glutathione was included in the test solutions. Increasing internal medium osmolarity and extrapolation to infinite osmolarity indicated that 50% of the measured complex in cLPM was due to nonspecific membrane binding. Uptake of the CH3HgSG complex was not affected by the replacement of a sodium-chloride gradient with gradients of potassium-chloride or lithium-chloride. Addition of ATP had no effect on CH3HgSG uptake. A significant increase in CH3HgSG uptake was detected when valinomycin was included in an external medium containing 100 millimolar (mM) potassium-gluconate. Uptake of tritiated glutathione (GSH) was also increased under these conditions. Concentration dependent increases in the initial rates of CH3HgSG uptake followed Michaelis-Menten kinetics for substrate concentrations from 5 micromolar to 5 millimolar. Uptake of the CH3HgSG complex was inhibited by GSH, its S-methyl, S-ethyl, S-butyl, S-hexyl, S-octyl, and S-dinitrophenyl conjugates as well as by ophthalmic acid. Uptake was not effected by oxidized glutathione (GSSG), bile acids, amino acids or P-glycoprotein inhibitors. Competitive inhibition of CH3HgSG uptake by GSH was demonstrated by kinetic studies. Preloading of cLPM with CH3HgSG or GSH significantly increased uptake of CH3HgSG. The authors conclude that CH3HgSG and GSH share canalicular transport carriers that are responsible for the elimination of methylmercury (22967926) via bile excretion.

Abstract  The present work consisted in the study of the oxidation Yield induced by catalysts during ozonation of sodium gluconate, toluene sulfonic acid and Triton X100 in dilute aqueous solution (TOC = 8 mg C/L). Two operating conditions have been used to demonstrate the enhancement brought about by catalytic ozonation: continuous filtration experiments and batch tests.

Dynamic filtration experiments have been performed on catalysts previously saturated to simulate the behavior of the catalyst over a long period of use.

Batch experiments which allowed us to distinguish the part due to adsorption, ozonation and catalytic effect were carried out on three catalysts with intent to test their efficiency. (C) 1998 Published by Elsevier Science Ltd. All rights reserved.

Abstract  BIOSIS COPYRIGHT: BIOL ABS. Aluminum (Al), one of the most widespread element on earth, often contaminates preparations such as parenteral nutrition solutions. Small-volume additives such as calcium gluconate and phosphate salts solutions are the most contaminated. Large-volume parental source solutions such as dextrose, crystalline amino acids and lipid emulsions are significantly less contaminated. Premature infants generally need intravenous feeding and are therefore vulnerable to aluminum toxicity: protective gastrointestinal mechanisms are bypassed and renal function is immature. Several studies showed that Al blood levels and urinary excretion increase when premature infants received parenteral nutrition. Urinary elimination is not adequate and Al accumulation is observed in tissues, especially in bones. Recently, in preterm infants, prolonged intravenous feeding with solutions containing Al was demonstrated to be associated with impaired neurologic development. Montreuil Hospital intensive c

Abstract  HEEP COPYRIGHT: BIOL ABS. NOTE HUMAN CALCIUM GLUCONATE ALUMINUM SALTS CALCIUM EDTA SERPENTINE BORON IRON METABOLIC-DRUG ANTIDOTE OSTEO POROSIS

Abstract  BIOSIS COPYRIGHT: BIOL ABS. A multinational interlaboratory study to investigate the bovine corneal opacity and permeability (BCOP) assay is presented. The aim of this work was to determine the capability and possible limitations of this method to predict ocular irritancy of a large set of chemicals. The assays were carried out in 12 European laboratories with different types of activity. In each of these laboratories 52 substances, with a wide range of structure, physical form and irritant properties, were tested and in vitro scores were compared with those obtained from concurrent rabbit eye (Draize) tests. The technique was easily learned by workers in the participating laboratories, as shown by the fact that there were consistent responses between treated corneas within an individual laboratory. Interlaboratory variability was also very good. It was found that a given laboratory had a 96% chance of classifying irritants or non-irritants similarly to the other laboratories. In addition, it was

Abstract  The aim of this work was to characterize and to quantify ozonation by-products of glucose in ultrapure water ([Glucose](o) = 5 mmol 1(-1); initial pH : pH approximate to 6, final pH : approximate to 3). HPLC and GC analyses showed that ozonation of glucose yields mainly gluconic acid (0.6 - 0.7 mole I mole of glucose eliminated) and formic acid (0.2 - 0.25 mole / mole of glucose eliminated) at the beginning of the reaction. Glycolic acid, formaldehyde and peroxides were also formed for low ozone dosages. GC/MS analyses carried out on extracts from ozonated solutions of glucose and gluconic acid (90-95% removal of the initial organic compound) allowed the identification of several by-products wih six carbon atoms (hexosuloses, glucuronic and glucaric acids) and with less carbon atoms (acids and aldehydes).