Tumoricidal activity of lauryl gallate towards chemically induced skin tumours in mice.

Lauryl gallate (antioxidant food additive E-312) prevents the formation of dimethylbenzanthracene-induced skin tumours in mice, and kills, selectively, tumoral cells on established tumours. This results in total remission, after topical application of the compound on the tumoral mass, without affecting the surrounding tissue.

In vitro toxicity of stimulant soft drinks.

Some drinks containing natural products with stimulant properties are highly consumed among young adults. The market for these drinks has increased in past years around the globe, and, although they might be harmless, overdoses or combination of these with other drinks could be harmful to the health of some consumers in certain circumstances. Samples were obtained at food shops and different popular brands were chosen. Cytotoxicity tests used were neutral red uptake, total protein content, and tetrazolium assay on Chinese hamster ovary cells. Results revealed that tested samples were not cytotoxic; however, studies have demonstrated the toxicity of high concentrations of some of these products. For this reason, the authors considered it to be of critical importance to carryout an in vitro toxicity screening of stimulant soft drinks that are highly consumed.

Sister chromatid exchange induced by several types of coffees in Chinese hamster ovary cells.

Different brands of commercial caffeinated and decaffeinated coffees (roasted, high roast, blend ground, and instant coffees) were studied. These coffees were tested for their ability to induce sister chromatid exchanges (SCE) in CHO-K1 cells. Tests were performed in the presence and in the absence of a metabolic activation system (S-9 mix). Results were compared to the roasting procedure because genotoxic products could be formed from these processes. Our results indicate that caffeinated instant coffees showed higher genotoxic activity than decaffeinated coffees. Non-significant genotoxic activity was detected with the green coffee (unroasted). The highest increase of the frequency of SCE occurred when the caffeinated instant coffee was tested in the absence of metabolic activation system. The repeatability of the test was checked through three assays with the same sample.

Pharmacokinetics of intravenous luxabendazole in rabbits: influence of the enterohepatic circulation.

Luxabendazole (LBZ) is a new benzimidazole carbamate chemotherapeutic agent, which has proved to be very effective against adult and immature stages of the major gastrointestinal nematodes, trematodes and cestodes. While information on the efficacy of LBZ in several animal species is available, there seems to be no published information describing the disposition kinetics in any of them. As a part of the clinical development of luxabendazole, the pharmacokinetics of a single intravenous dose was investigated in parasite-free rabbits. Serial blood samples were collected at timed intervals for 12 h following administration of the dose, and concentrations in plasma were determined by a sensitive and specific HPLC method. Published data on LBZ point to the possible existence of an enterohepatic cycle (EHC), and so, it seemed appropriate to carry out two different forms of test. In the first, the possibility of intestinal reabsorption of LBZ excreted via the bile was allowed for (Treatment 1), while in the second it was interrupted by the oral administration of activated charcoal (Treatment 2). In both cases the animals were given a single dose of 10 mg kg-1 of LBZ intravenously (i.v). Comparison of the areas under the curve (AUCs) of LBZ concentrations in plasma samples taken from the animals receiving each treatment showed significant difference (p < 0.05). The given dose (10 mg kg-1) was converted in Treatment 1 to an effective dose of 13.9 mg kg-1 through recycling of LBZ. With Treatment 2 a bicompartmental distribution model for this drug was confirmed, together with high apparent distribution volumes: Vc = 1.87 L kg-1, and V beta = 7.09 L kg-1.

Pharmacokinetics of luxabendazole after oral and intravenous administration to sheep.

OBJECTIVE: To determine pharmacokinetics of luxabendazole after oral and i.v. administration to healthy sheep. ANIMALS: 7 clinically normal female Merino sheep between 9 and 12 months old. PROCEDURE: Pharmacokinetics were determined after oral and i.v. administration of luxabendazole at a dose of 10 mg/kg of body weight. Serial blood samples were collected for 56 hours after administration. Plasma concentrations of luxabendazole were determined by high-pressure liquid chromatography. RESULTS: After i.v. administration, elimination of luxabendazole was slow, with a mean half-life of 8.72 hours. Mean steady-state volume of distribution and mean distribution volume during the elimination phase were 3.18 and 3.10 L/kg, respectively. Mean clearance was 0.24 L/kg.h, and mean area under the concentration-time curve was 41.89 mg.h/L. After oral administration, luxabendazole was slowly absorbed from the gastrointestinal tract. Mean absorption half-life was 2.26 hours. Peak plasma concentration was 0.50 microgram/ml and was detected 14 to 16 hours after drug administration. Mean area under the concentration-time curve was 12.03 mg.h/L. Mean bioavailability was 29%. CONCLUSIONS: Results suggest that luxabendazole is moderately absorbed from the gastrointestinal tract in sheep, is widely distributed into extravascular compartments, and is cleared slowly. CLINICAL RELEVANCE: Determination of pharmacokinetic parameters is the first step in determining a safe and efficacious dosage regimen for luxabendazole in sheep.

Determination of pentamidine in serum and urine by micellar electrokinetic chromatography.

A number of parameters influencing the electrokinetic processing of pentamidine by micellar electrokinetic chromatography (MEKC) were studied in order to develop an analytical method for this compound. The parameters considered were: pH, ionic strength, and SDS concentration of electrolyte, temperature and working voltage. On the basis of the results obtained, the best analytical conditions for the detection of pentamidine in serum and urine by MEKC were determined. Analysis by MEKC permitted determination of the drug in 10 min. Good linearity, reproducibility and accuracy were obtained in the range 0-30 micrograms/ml for both samples, with a correlation coefficient r > or = 0.9998 and a recovery of 87-92% in serum and 90-108.9% in urine. We examined the metabolism of pentamidine using rat liver homogenates in order to exclude any possible interference of metabolites in the analysis of pentamidine.

Bacterial mutagenic evaluation of Luxabendazole, a new broad spectrum anthelmintic, with the Salmonella typhimurium His- and the Escherichia coli Tryp- reversion tests.

Luxabendazole is a new benzimidazole carbamate chemotherapeutic agent, which has proved to be effective against adult and immature stages of the major gastrointestinal nematodes, trematodes and cestodes. The mutagenic properties of Luxabendazole were investigated in the in vitro Ames Salmonella and E. coli tests. The product was tested at concentrations of 0.5, 5, 50, 500, 1250 and 2500 micrograms/plate in the TA1535, TA1538, TA98 and TA100 strains of Salmonella typhimurium, and 0.5, 5, 50 and 500 micrograms/plate in the WP2, WP2uvrA- and its pKM 101-containing derivative CM891 (WP2 uvrA- pKM101) strains of Escherichia coli, with and without S9 microsomal activation (post-mitochondrial liver fraction from Wistar rats pretreated with Aroclor(R)). Positive and negative controls were included in each experiment. From the present study it can be concluded that Luxabendazole, over a dose range of 0.5-2500 micrograms/plate, is unlikely to present a mutagenic hazard, as demonstrated by the Ames test.

Pharmacokinetics of triclabendazole in rabbits.

1. Pharmacokinetic profiles of triclabendazole (TCBZ) following intravenous (i.v.) and oral administration of the drug in rabbits were carried out. 2. In normal rabbits, TCBZ was metabolized rapidly to its sulphoxide (TCBZ-SO) and sulphone (TCBZ-SO2) derivatives following administration, with undetectable concentrations of unchanged TCBZ in the plasma of the treated animals at any time (detection limit, 10 ng/ml). 3. The disposition kinetics of this drug in rabbits can be described by a two-compartment open model. 4. Mean peak concentrations in plasma of TCBZ-SO and TCBZ-SO2 of 12.41 micrograms/ml and 9.5 micrograms/ml occurred 7.5 and 9.5 hr after oral administration, respectively. 5. Both metabolites were eliminated slowly from plasma with elimination half-lives of 16.86 hr for the sulphoxide and 13 hr for the sulphone. 6. The area under the plasma concentration versus time curve (AUC) was 240 mg hr/l for the sulphoxide, higher than that found for the sulphone, 185 g hr/l.

Aromatic diamidines are reversible inhibitors of porcine kidney diamine oxidase.

The inhibitory ability of aromatic diamidines has been studied on porcine kidney diamine oxidase. The reversibility of drug-protein interactions has been tested by means of exhaustive dialysis experiments, showing in all cases a reversible binding pattern. Ki values obtained by means of Lineweaver-Burk plots were: stilbamidine 12 microM, 2-OH-stilbamide 8.5 microM, phenamidine 4 microM, propamidine 8 microM, dibromopropamidine 4.9 microM and amicarbalide 12 microM.

Reversed-phase ion-pair high-performance liquid chromatographic determination of triclabendazole metabolites in serum and urine.

An ion-pair high-performance liquid chromatographic method was developed for measuring the concentrations of triclabendazole metabolites (sulphoxide and sulphone) in plasma and urine samples. The diluted biological fluids are ultrafiltered before chromatography through a 30,000 relative molecular mass cut-off filter and then injected into a C18 column. They are then isocratically eluted with a mobile phase consisting of 0.05 M phosphate buffer (pH 7.0)-acetonitrile (55:45, v/v) with addition of 1.0 mmol/l sodium decanesulphonate and monitored by ultraviolet-visible spectrophotometry at 312 nm. Recoveries over the range 0.01-9.0 micrograms/ml for triclabendazole sulphoxide and sulphone are, respectively, 91.7% and 91.6% in serum and 90.3% and 90.2% in urine. For both metabolites, the limit of detection is 10 ng/ml in both urine and serum.

Inhibition of diamine oxidase from porcine kidney by pentamidine and other aminoguanidine compounds.

1. Three bisguanidine compounds (those of pentamidine, streptidine and phenformin) were compared for their in vitro inhibitory capacity on diamine oxidase activity (EC 1.4.3.6), the first enzyme of putrescine degradation. 2. Pentamidine was the most potent inhibitor, and phenformine the weaker. Two and a half micromoles of pentamidine was enough to reduce the enzyme activity by 50%, while streptidine and phenformin produced the same effect at concentrations greater than 0.90 and 4 mM, respectively. 3. Pentamidine, streptidine and phenformin appeared to be non-competitive inhibitors, and the Ki values calculated by a Dixon plot were 3 microM, 0.95 mM and 4 mM, respectively.

In vitro synthesis of colominic acid by membrane-bound sialyltransferase of Escherichia coli K-235. Kinetic properties of this enzyme and inhibition by CMP and other cytidine nucleotides.

The membrane-bound sialyltransferase obtained from Escherichia coli K-235 grown in a chemically defined medium (ideal for colominic acid production) was studied. The in vivo half-life calculated for this enzyme was 20 h. Kinetic tests revealed (at 33 degrees C and pH 8.3) hyperbolic behaviour with respect to CMP-Neu5Ac (Km250 microM) and a transition temperature at 31.3 degrees C. The enzyme was inhibited by NH4+, some divalent cations and by several agents that react with thiol groups. Detergents and fatty acids also inhibited the sialyltransferase activity. In vitro synthesis of colominic acid is strongly inhibited by CMP by blocking the incorporation of [14C]Neu5Ac into a protein-complex intermediate and therefore into free polymer. CDP and CTP also inhibited (91% and 84%) this enzyme activity whereas cytosine and cytidine had no effect. CMP inhibition corresponded to a competitive model the calculated Ki was 30 microM. Incubations of protein[14C]Neu5Ac with CMP, CDP and CTP led to de novo synthesis of CMP-[14C]Neu5Ac. The presence of colominic acid, which usually displaces the reaction equilibrium towards polymer synthesis, did not affect this de novo CMP-[14C]Neu5Ac formation. CMP also inhibited in vivo colominic acid biosynthesis.

A protein-sialyl polymer complex involved in colominic acid biosynthesis. Effect of tunicamycin.

A protein-NeuAc complex involved in colominic acid biosynthesis has been identified in membrane preparations of Escherichia coli K-235. This compound had an Mr (estimated by SDS/polyacrylamide-gel electrophoresis and autoradiography) of about 100,000 and played the role of an 'initiator' or 'primer' (endogenous acceptor) in the synthesis of the whole polymer. Incubations of E. coli membranes with CMP-[14C]NeuAc (CMP-N-[14C]acetylneuraminic acid) pointed to the existence of a protein fraction (primer acceptor) that linked residues of sialic acid (N-acetylneuraminic acid, NeuAc) up to a maximal size, later releasing them as low-Mr sialyl polymers (LMrS, Mr less than 10,000). In the presence of colominic acid (final acceptor) the radioactivity linked to the protein quickly decreased, appearing stoichiometrically bound to the whole polysaccharide. When membrane preparations were previously digested with Streptomyces proteinase or de-activated by heating (80 degrees C, 10 min), no incorporation of labelled NeuAc into trichloroacetic acid-insoluble material was detected. These results suggested that colominic acid molecules are synthesized while they are bound to a proteinaceous acceptor that is subsequently excised in the presence of colominic acid, generating the native protein. The antibiotic tunicamycin inhibited the biosynthesis of colominic acid, affecting the synthesis of this protein-(NeuAc)n intermediate. All these results are described here for the first time.