Antimutagenic activity of natural phenolic compounds present in the common bean ( Phaseolus vulgaris ) against aflatoxin B 1

Polyphenols with antimutagenic and anticarcinogenic properties are present in fruits, vegetables and legumes. In this study, the Salmonella typhimurium tester strains TA98 and TA100 were used in the microsuspension assay to examine the antimutagenic effect of phenolic compounds extracted from the common bean (Phaseolus vulgaris) against mutagenicity induced by aflatoxin B 1 (AFB 1 ). A dose-response curve was constructed for AFB 1 ; from which a level of 40ng AFB 1 /tube was selected for all antimutagenicity assays. The AFB 1 and phenolic extract (PE) were not toxic to the bacteria at concentrations tested. In the case of PE, results were similar to the number of spontaneous revertants for TA98 and TA100. The inhibitory effect of PE against AFB 1 mutagenicity was dose-dependent at the lower concentrations tested (2.5, 5, 10, 12.5, 15 and 25 μg-equivalent ( + )-catechin/tube for TA98; 0.5, 1, 1.5, 2.5, 5, 10 and 25 μg-equivalent ( + )-catechin/ tube for TA100). Further, a two-stage incubation procedure was used to investigate the potential interaction between PE and AFB 1 . The greatest inhibitory effect of the PE on AFB 1 mutagenicity occurred when PE and AFB 1 were incubated together. When the bacteria were first incubated with PE followed by a second incubation with AFB 1 , lower inhibition was observed. Lower inhibition was also observed when the bacteria were first incubated with AFB 1 followed by a second incubation with PE. The results suggest that the mechanism of inhibition could involve the formation of a chemical complex between of PE and AFB 1 .     

Mechanical improvement of hydroxyapatite by TiO<Subscript><Emphasis Type="Italic">x</Emphasis></Subscript> nanoparticles deposition

Deposition of Ti was carried out by laser ablation onto hydroxyapatite porous discs in an Ar atmosphere. Ti nanoparticles were deposited onto HAp surface in order to modify its roughness and morphology as it is observed by scanning electron microscopy (SEM) and scanning probe microscopy (SPM). A homogeneous distribution of Ti over the disc surface was corroborated by elemental mapping. A comparison of the hydroxyapatite hardness before and after deposition was performed using SPM nanoindentation. Transmission Electron Microscopy (TEM) showed that the Ti nanoparticles obtained were covered by an oxygen shell. It is shown that surface modifications of the covered HAp by Ti result in better mechanical properties, reducing the possible damage to the HAp ceramic by friction or impacts as it often happens in meniscus, bone junctions and the inclusion of prosthesis for human treatments.     

Transport of chromium(VI) through a Cyanex 921‐supported liquid membrane from HCl solutions

The transport of chromium(VI) through a flat‐sheet supported liquid membrane containing Cyanex 921 as a carrier has been investigated. The permeation of the metal is investigated as a function of various experimental variables: hydrodynamic conditions, concentration of chromium(VI) and HCl in the feed phase, carrier concentration and diluent in the membrane and strippant concentration in the stripping phase. The mass transfer coefficient and the thickness of the aqueous boundary layer were calculated from the experimental data. Furthermore, the selectivity of Cyanex 921‐based flat‐sheet supported liquid membrane towards different metal ions and the behaviour of the system against other carriers are presented. Copyright © 2003 Society of Chemical Industry     

Synthesis and release of ATP by soluble mitochondrial F1 in complex with its inhibitor protein during dimethylsulfoxide-water transitions

Soluble mitochondrial F1 and F1 in complex with the natural ATPase inhibitor protein (F1-IP) catalyze the spontaneous synthesis of [gamma-32P]ATP from medium [32P]phosphate and enzyme-bound ADP when incubated in media with dimethylsulfoxide (Me2SO); under these conditions, the synthesized [gamma-32P]ATP is not released into the media, it remains tightly bound to the enzymes [Gómez-Puyou, A., Tuena de Gómez-Puyou, M. & de Meis, L. (1986) Eur. J. Biochem. 159, 133-140]. Some of the characteristics of the synthesized [gamma-32P]ATP were studied in F1 and F1-IP (ATPase activities of 70 and 1-3 micromol x min(-1) x mg(-1), respectively). In Me2SO media, gamma-phosphate of synthesized ATP in F1 or F1-IP exchanges with medium phosphate. From the rates of the exchange reaction, the half-times for hydrolysis of the synthesized ATP in F1 and F1-IP were calculated: 45 min and 58 min for F1 and F1-IP, respectively. The course that synthesized [gamma-32P]ATP follows after dilution of the Me2SO synthetic mixture with aqueous buffer was determined. After dilution, the half-life of synthesized ATP in F1 was less than 1 min. In F1-IP, ATP was also hydrolyzed, but at significantly lower rates. In F1-IP, dilution also produced release of the synthesized [gamma-32P]ATP. This was assayed by the accessibility of [gamma-32P]ATP to hexokinase. About 25% of [gamma-32P]ATP synthesized in F1-IP, but not in F1, was released into the media after dilution with aqueous buffer that contained 20 mM phosphate. Release of tightly bound ATP required the binding energy of phosphate and solvation of F1-IP, however, the particular kinetics of F1-IP were also central for medium ATP synthesis in the absence of electrochemical H+ gradients.     

Cellular UDP-glucose deficiency caused by a single point mutation in the UDP-glucose pyrophosphorylase gene

We previously isolated a mutant cell that is the only mammalian cell reported to have a persistently low level of UDP-glucose. In this work we obtained a spontaneous revertant whose UDP-glucose level lies between those found in the wild type and the mutant cell. The activity of UDP-glucose pyrophosphorylase (UDPG:PP), the enzyme that catalyzes the formation of UDP-glucose, was in the mutant 4% and in the revertant 56% of the activity found in the wild type cell. Sequence analysis of UDPG: PP cDNAs from the mutant cell showed one missense mutation, which changes amino acid residue 115 from glycine to aspartic acid. The substituted glycine is located within the largest stretch of strictly conserved residues among eukaryotic UDPG:PPs. The analysis of the cDNAs from the revertant cell indicated the presence of an equimolar mixture of the wild type and the mutated mRNAs, suggesting that the mutation has reverted in only one of the alleles. In summary, we demonstrate that the G115D substitution in the Chinese hamster UDPG:PP dramatically impairs its enzymatic activity, thereby causing cellular UDP-glucose deficiency.     

Characterization of a lectin-like protein isolated from Lachesis muta snake venom

A lectin-like protein was isolated from L. muta venom by gel filtration on BIO Gel P-100 followed by column Chromatography on DEAE-sephades A-50. The protein eluted at 0.4 M Nacl in 0.01 Tris pH 7.3 and exhibited agglutinin activity toward 0+ human erythrocytes. The protein is a dimer with Mr 28 kDa. Amino acid analysis revealed high content of tryptophan and acid recidues and low content of cysteine and methionine residues. No neutral carbohydrates and sialic acid were detected. Circular dichroic spectrum shows 78% of B structure and 1% of alpha structure. In vitro experiments with erythrocytes from rat, rabbit and dog revealed strong agglutination while red blood cells from mice, sheep and goat were not agglutinated. In vivo experiments using anesthetized rats, a sharp and prolonged fall in the blood pressure was observed at protein dose of 1.5 mg/kg. Double dose of protein caused the death of the animal.