Antimutagenicity of supercritical CO2 extracts of Terminalia catappa leaves and cytotoxicity of the extracts to human hepatoma cells

Natural antimutagens may prevent cancer and are therefore of great interest to oncologists and the public at large. Phytochemicals are potent antimutagen candidates. When the Ames test was applied to examine the antimutagenic potency of supercritical carbon dioxide (SC-CO(2)) extracts of Terminalia catappa leaves at a dose of 0.5 mg/plate, toxicity and mutagenicity were not detected. The antimutagenic activity of SC-CO(2) extracts increased with decreases of temperature (60, 50, and 40 degrees C) and pressure (4000, 3000, and 2000 psi) used for extraction. The most potent antimutagenicity was observed in extracts obtained at 40 degrees C and 2000 psi. At a dose of 0.5 mg of extract/plate, approximately 80% of the mutagenicity of benzo[a]pyrene (B[a]P, with S-9) and 46% of the mutagenicity of N-methyl-N '-nitroguanidine (MNNG, without S-9) were inhibited. Media supplemented with SC-CO(2) extracts at a range of 0-500 microg/mL were used to cultivate human hepatoma (Huh 7) and normal liver (Chang liver) cells. The viability of the cells was assayed by measuring cellular acid phosphatase activity. A dose-dependent growth inhibition of both types of cells was observed. The SC-CO(2) extracts were more cytotoxic to Huh 7 cells than to Chang liver cells. The observation that SC-CO(2) extracts of T. catappa leaves did not induce mutagenicity at the doses tested while exhibiting potent antimutagenicity and were more cytotoxic to human hepatoma cells than to normal liver cells is of merit and warrants further investigation.     

Differential Tolerance to Lead and Cadmium of Micropropagated <Emphasis Type="Italic">Gypsophila fastigiata</Emphasis> Ecotype

In vitro techniques may provide a suitable tool for effective propagation and conservation of plant species representing various ecological niches. The elaboration of such protocols is also prerequisite for selection of heavy-metal-tolerant plant material that could be afterwards used for restoration or remediation of polluted sites. In this study, culture protocol for Gypsophila fastigiata propagation was developed. The highest multiplication coefficient, which reached 6.5, and the best growth parameters were obtained on modified MS medium supplemented with 1.0 mg L^?1 2iP and 0.2 mg L^?1 IAA. The obtained cultures were treated with different concentrations of lead nitrate (0.1, 0.5, and 1.0 mM Pb(NO₃)₂) or cadmium chloride (0.5, 2.5, and 5.0 μM CdCl₂). The growth parameters, photosynthetic pigments, and phenolic compound content were examined in order to evaluate whether tested metal salts can have an adverse impact on studied culture. It was ascertained that Pb ions induced growth disturbances and contributed to shoot wither. On the contrary, the proliferative shoot cultures were established on media containing Cd ions and the multiplication coefficients and shoot length increased on all media enriched with CdCl₂. Chlorophylls and carotenoid contents were negatively affected by application of 5.0 μM of cadmium; nevertheless, in shoots treated with 2.5 μM CdCl₂, increased accumulation of photosynthetic pigments occurred and their amount was similar to untreated culture. Adaptation to Cd was associated with stimulation of phenolic compound synthesis. Hence, we have reported on unambiguous positive result of in vitro selection procedure to obtain vigorous shoot culture tolerant to cadmium.     

Germination of peanut kernels to enhance resveratrol biosynthesis and prepare sprouts as a functional vegetable

Bioactive benefits of resveratrol in the diets have attracted extensive interests of the public. Peanut is one of the potent natural sources of resveratrol. In this study, germination of peanut kernels to enhance resveratrol biosynthesis and preparation of sprouts as a functional vegetable was conducted. When the rehydrated kernels of three peanut cultivars were germinated at 25 degrees C and relative humidity 95% in dark for 9 days, resveratrol contents increased significantly from the range of 2.3 to 4.5 microg/g up to the range of 11.7 to 25.7 mug/g depending upon peanut cultivar. In comparison with the sprout components, resveratrol contents were highest in the cotyledons, slightly lower in the roots, and not detected in the stems. When the sprouts were heated in boiling water for 2 min, resveratrol contents varied in a limited range. Methanol extracts of the freeze-dried sprouts exhibited potent 1,1-diphenyl-2-picryl-hydrazyl scavenging activity and antioxidative potency against linoleic acid oxidation. These activities increased with an increase of germination time. After 9 days of germination, total free amino acid, sucrose, and glucose contents increased significantly while crude protein contents decreased and the large sodium dodecyl sulfate polyacrylamide gel electrophoresis protein molecules of the kernels were extensively degraded. From a practical viewpoint, it is of potency to prepare peanut sprouts as a functional vegetable.     

Development of an immunoassay for the residues of the herbicide bensulfuron-methyl

To develop a competitive indirect enzyme-linked immunosorbent assay based on polyclonal antibodies for the detection of the sulfonylurea herbicide bensulfuron-methyl, seven structurally related haptens were synthesized. Four of them mimicking the target analyte were conjugated to keyhole limpet hemocyanin by the N-hydroxysuccinimide activated ester method to use as immunogens, and all of them were conjugated to bovine serum albumin to use as plate-coating antigens. Polyclonal antibodies raised in rabbits and the coating antigens were screened and selected for the assay in simple homologous and heterologous ELISA formats. Three sensitive heterologous ELISAs were selected and optimized, showing the average IC(50) values of bensulfuron-methyl as low as 0.17, 0.09, and 0.09 ng/mL, the detection ranges of 0.04-0.60, 0.01-0.60, and 0.04-0.25 ng/mL, and the lowest detection limits of 0.03, 0.002, and 0.03 ng/mL, respectively. The cross-reactivities of other sulfonylurea herbicides and metabolites of bensulfuron-methyl to the antibodies were less than 15% in the two assays. Recoveries from the analyte-fortified water samples in assay I were in the range of 81-125% by simple dilution. The correlation between the ELISA and HPLC was 0.999 (n = 15) with a slope of 1.37 in the analysis of groundwater samples fortified with bensulfuron-methyl. The results obtained strongly indicate that the ELISA can be a highly sensitive and convenient tool for detecting bensulfuron-methyl residues in agricultural and environmental samples.     

Physiological response of culture media-grown barley (Hordeum vulgare L.) to titanium oxide nanoparticles

Since the fate of nanoparticles after their release in the environment and their possible transfer in plants and subsequent impacts is still largely unknown, this paper evaluates the potential phytotoxic effects of up to 20% w/w TiO₂ nanoparticles (nTiO₂) on barley cultivated in hydroponics and agar media. The X-ray diffraction analysis confirmed that nTiO₂ powder corresponds to anatase phase. On agar medium only high concentrations of nTiO₂ (10% and 20% w/w) induced significant inhibition of shoot growth. However, hydroponics treatment with nTiO₂ up to 1000?mg?L^?1 did not show any adverse effect on the shoot growth. In both experiments, (i) root growth inhibition effects became visible with increasing concentration of nTiO₂, (ii) plants treated with nTiO₂ showed no change in chlorophyll a and b content, even though the plants absorbed nTiO₂, (iii) weight of biomass treated with nTiO₂ was not significantly different compared to control. Therefore, we assume that transport of nTiO₂ into the aerial parts is limited due to the presence of effective mechanical or physiological barriers in roots. Overall, it appears that early root growth is a relevant indicator of potential effects of nTiO₂ exposure. Our results also indicate that synthesized nTiO₂ are not significantly toxic to the barley when applied at the concentrations used in this work, even though plants absorb titanium.     

Degradation of Organic Compounds in Actual Wastewater by Electro-Fenton Process and Evaluation of Energy Consumption

We investigated the Fe-regeneration-based cathodic electro-Fenton process using commercial electrodes, to achieve reducing the initial Fe dose and consequent sludge accumulation. The obtained results provided insights into the cathodic electro-Fenton process using two-electrode electrochemical system. The grade 2 Ti mesh electrode could effectively regenerate Fe(II) at 1.2 V, whose potential was lower than those of the two other electrode systems reported in the literatures. In the actual wastewater treatment, compared with general-Fenton process, the reduction in the initial Fe injection was 33% and the sludge reduction was 38%. Furthermore, the removal efficiency using electro-Fenton was at least 5% higher than that of the general-Fenton process at the same initial dose. The low cell voltage (1.2 V) led to lower energy consumption (3.5 J mg^?1) relative to those of other electro-Fenton systems, i.e., anodic electro-Fenton and peroxide generation. With its competitive degradation performances, low energy consumption, and reduced initial Fe injection, the present cathodic electro-Fenton process was shown to be a promising route of waste treatment.     

RESPONSE OF ‘GRANNY SMITH’ APPLE TREES TO FOLIAR TITANIUM SPRAYS UNDER CONDITIONS OF LOW SOIL AVAILABILITY OF IRON,MANGANESE, AND ZINC

The aim of the study was to examine impact of foliar titanium (Ti) sprays on vegetative and reproductive response of apple (Malus domestica Borkh.) trees under conditions of low soil availability of iron (Fe), manganese (Mn), and zinc (Zn). The experiment was conducted during 2005–2006 at a Experimental Station in Isparta region, Turkey, on mature ‘Granny Smith’ apple trees/M.9, planted at a spacing of 3.5 × 1.5 m, on fine-textured soil with neutral reaction, medium status of organic matter, high amounts of available phosphorus (P), potassium (K), and magnesium (Mg), and low availability of Fe, Mn, and Zn. The trees were sprayed with Ti-ascorbate at the green and pink bud stage, petal fall, and 3, 6, and 9 weeks after full bloom, at a rate of 3 g Ti ha^?1 per spray. The efficiency of Ti sprays was compared to combined sprays of Fe, Mn, and Zn [chelated with ethylenediaminetetraacetic acid (EDTA)], applied at the same terms as Ti sprays, at rate of 36 g, 36 g and 24 g per spray, respectively. Trees unsprayed with Ti, Fe, Mn, and Zn served as the control. It was shown that summer leaf Ti concentrations of the trees untreated with Ti were high, varying from 34 to 36 mg kg^?1 dry matter. Foliar Ti sprays increased leaf status of this nutrient but they had no effect on nutrition of essential macro- and microelements, tree vigor, and fruit yield. Mean apple weight, coloring, firmness, soluble solids concentration, and titratable acidity of fruit were not also influenced by Ti sprays. Foliar sprays of Fe, Mn, and Zn improved leaf status of nitrogen (N), Mg, Fe, Mn, and Zn; leaves of the trees sprayed with those micronutrients were also greener, and contained more Fe²⁺ than those of the control plants. Combined sprays of Fe, Mn, and Zn improved tree vigor and fruit yield.     

All hermaphrodite progeny are derived by self-pollinating the sunrise papaya mutant

Self‐pollination of a hermaphroditic cultivar normally gives a ratio of 2 : 1 hermaphrodite to female papayas with genotypes M₂m and mm, respectively. Much effort has been dedicated to marking the sexual types of papaya at the seedling stage to distinguish hermaphroditic from female papayas. A hermaphroditic papaya mutant (SR*) has been obtained, derived from the ‘Sunrise’ papaya cultivar mutant. Self‐pollination of the mutant resulted in all progenies being hermaphroditic. The genotype of the female was lethal, as a result of a lethal gene being linked to the mm female gene complex in this case. However, a 3 : 1 segregation ratio was obtained from the progeny of the hermaphroditic cultivar ‘Thailand’ crossed with SR*, indicating that all genotypes survived. Homozygous genotypes (M₂M₂) would be lethal according to Storey's model. Randomly selected F1 plants of the ‘Thailand’ SR* combination were self‐pollinated to obtain an F₂ generation. The F₂ segregation ratio suggested that the SR* mutant had a different form of the M₂ allele, now designated as M_@, which allowed the dominant M_@M₂ to survive in cross combinations. Genetic study has proved that SR* has the M_@ml genotype, a new mutant. It is capable of producing all hermaphroditic papaya progenies.     

Rapid pneumococcal evolution in response to clinical interventions

Epidemiological studies of the naturally transformable bacterial pathogen Streptococcus pneumoniae have previously been confounded by high rates of recombination. Sequencing 240 isolates of the PMEN1 (Spain(23F)-1) multidrug-resistant lineage enabled base substitutions to be distinguished from polymorphisms arising through horizontal sequence transfer. More than 700 recombinations were detected, with genes encoding major antigens frequently affected. Among these were 10 capsule-switching events, one of which accompanied a population shift as vaccine-escape serotype 19A isolates emerged in the USA after the introduction of the conjugate polysaccharide vaccine. The evolution of resistance to fluoroquinolones, rifampicin, and macrolides was observed to occur on multiple occasions. This study details how genomic plasticity within lineages of recombinogenic bacteria can permit adaptation to clinical interventions over remarkably short time scales.     

Loss of imprinting of Igf2 alters intestinal maturation and tumorigenesis in mice

Loss of imprinting (LOI) of the insulin-like growth factor II gene (IGF2) is an epigenetic alteration that results in a modest increase in IGF2 expression, and it is present in the normal colonic mucosa of about 30% of patients with colorectal cancer. To investigate its role in intestinal tumorigenesis, we created a mouse model of Igf2 LOI by crossing female H19+/- mice with male Apc+/Min mice. Mice with LOI developed twice as many intestinal tumors as did control littermates. Notably, these mice also showed a shift toward a less differentiated normal intestinal epithelium, reflected by an increase in crypt length and increased staining with progenitor cell markers. A similar shift in differentiation was seen in the normal colonic mucosa of humans with LOI. Thus, altered maturation of nonneoplastic tissue may be one mechanism by which epigenetic changes affect cancer risk.