Ultrasensitive quantitative determination of paraquat: application to river, ground, and drinking water analysis in an agricultural area

The water specimens were collected from wells and irrigation ditches in the agricultural area to the south of Milan and from Olona River and Mantua Lake and analyzed for paraquat detection. The assay was performed using a specific polyclonal antibody raised in sheep and rabbit anti-sheep IgG conjugated with a chelating molecule 4,7-bis(chlorosulfophenyl)-1,10-phenanthroline-2,9-dicarboxylic acid complexed with Eu3+ as a fluorescent marker. Bovine serum albumin conjugated with 5-(1'-methyl[4,4']bipyridinyl-1-yl)pentanoic acid was used in solid phase preparation. The sensitivity achieved was 20 ng L(-1). The recovery in samples spiked with three different PQ concentrations was between 88 and 108%.     

A long-term in vitro culture of bovine epithelial cells on collagen rafts

In the present work, we established and characterized a 3D functional polarized primary bovine oviduct epithelial cells (BOECs) culture on free-floating type I collagen hydrogels (rafts) at an air-liquid interface (ALI). Intercellular junctions, ultrastructural cellular morphology and the expression of the OVGP1 closely recapitulated those of the in vivo epithelium lining. These morphological and physiological epithelial cell features were maintained under standard DMEM/F12 with 10% foetal bovine serum culture medium for at least 28 days of ALI culture. The versatility of the BOECs raft cultures should allow testing of toxicity compounds, in vitro evaluation of physiological or pathological oviductal states, and the study of epithelial-mesenchymal interactions that are critical for the maintenance of oviductal homeostasis.     

trans-Resveratrol, quercetin, (+)-catechin, and (-)-epicatechin content in south Italian monovarietal wines: relationship with maceration time and marc pressing during winemaking

The concentrations of trans-resveratrol, (+)-catechin, (-)-epicatechin, and quercetin were evaluated by means of high-performance liquid chromatography-diode array detection in red wines obtained from Aglianico, Piedirosso, and Nerello Mascalese grapes. The trans-resveratrol and epicatechin concentrations did not differ significantly between experimental wines. The concentration of quercetin in Nerello Mascalese wines was more than twice that observed in Aglianico and Piedirosso wines. Nerello Mascalese wines also significantly differed from other wines in the (+)-catechin content, which was significantly higher than those found in the other two wines. During maceration, the maximum extraction of trans-resveratrol was reached after 12 days for Aglianico and Piedirosso, after which a decline was observed. On the contrary, in the case of Nerello Mascalese, the concentration of trans-resveratrol increased steadily throughout the whole maceration process. After 2 days of maceration, the maximum concentration of quercetin was observed in Aglianico must, whereas the maximum quercetin extraction was reached after 12 days for Piedirosso and 17 days for Nerello Mascalese. The maximum levels of (+)-catechin and (-)-epicatechin were generally observed after 12 days of maceration for all wines, although a decline of (-)-epicatechin occurred after maximum extraction in Aglianico and Piedirosso wines. Following marc pressing, a significant increase in the concentration of trans-resveratrol for Aglianico, (+)-catechin and (-)-epicatechin for Piedirosso, and (-)-epicatechin for Nerello Mascalese was observed.     

Macromolecular architecture in eukaryotic cells visualized by cryoelectron tomography

Electron tomography of vitrified cells is a noninvasive three-dimensional imaging technique that opens up new vistas for exploring the supramolecular organization of the cytoplasm. We applied this technique to Dictyostelium cells, focusing on the actin cytoskeleton. In actin networks reconstructed without prior removal of membranes or extraction of soluble proteins, the cross-linking of individual microfilaments, their branching angles, and membrane attachment sites can be analyzed. At a resolution of 5 to 6 nanometers, single macromolecules with distinct shapes, such as the 26S proteasome, can be identified in an unperturbed cellular environment.     

Effectiveness of donor natural killer cell alloreactivity in mismatched hematopoietic transplants

T cells that accompany allogeneic hematopoietic grafts for treating leukemia enhance engraftment and mediate the graft-versus-leukemia effect. Unfortunately, alloreactive T cells also cause graft-versus-host disease (GVHD). T cell depletion prevents GVHD but increases the risk of graft rejection and leukemic relapse. In human transplants, we show that donor-versus-recipient natural killer (NK)-cell alloreactivity could eliminate leukemia relapse and graft rejection and protect patients against GVHD. In mice, the pretransplant infusion of alloreactive NK cells obviated the need for high-intensity conditioning and reduced GVHD. NK cell alloreactivity may thus provide a powerful tool for enhancing the efficacy and safety of allogeneic hematopoietic transplantation.     

Cilia-like beating of active microtubule bundles

The mechanism that drives the regular beating of individual cilia and flagella, as well as dense ciliary fields, remains unclear. We describe a minimal model system, composed of microtubules and molecular motors, which self-assemble into active bundles exhibiting beating patterns reminiscent of those found in eukaryotic cilia and flagella. These observations suggest that hundreds of molecular motors, acting within an elastic microtubule bundle, spontaneously synchronize their activity to generate large-scale oscillations. Furthermore, we also demonstrate that densely packed, actively bending bundles spontaneously synchronize their beating patterns to produce collective behavior similar to metachronal waves observed in ciliary fields. The simple in vitro system described here could provide insights into beating of isolated eukaryotic cilia and flagella, as well as their synchronization in dense ciliary fields.     

Abiotic forcing of plankton evolution in the Cenozoic

We characterize the evolutionary radiation of planktic foraminifera by the test size distributions of entire assemblages in more than 500 Cenozoic marine sediment samples, including more than 1 million tests. Calibration of Holocene size patterns with environmental parameters and comparisons with Cenozoic paleoproxy data show a consistently positive correlation between test size and surface-water stratification intensity. We infer that the observed macroevolutionary increase in test size of planktic foraminifera through the Cenozoic was an adaptive response to intensifying surface-water stratification in low latitudes, which was driven by polar cooling.     

Effects of Genotype and Environment on Phenolic Acids Content and Total Antioxidant Capacity in Durum Wheat

In cereals, phenolic acid (PA) content and total antioxidant capacity (TAC) may have a wide range of variability, probably because of several factors influencing the occurrence of grain antioxidants, which include genotype, environment, and their possible interactions. However, only a few studies have investigated the influence of these factors on durum wheat. In the present study, we investigated the impact of the genetic and environmental factors on the profile and content of PAs occurring as soluble free, soluble conjugated, and insoluble bound compounds, as well as on the TAC level, in three genotypes of durum wheat grown in three different Italian agroclimatic areas during two crop years. The results show that genotype, environment, and crop year have highly significant effects on TAC levels and on PA contents. In particular, TAC and free PAs are most influenced by year, whereas conjugated and bound PAs are most influenced by environment × year and genotype, respectively. Therefore, it is evidenced that genetic and environmental factors affect the antioxidant activity and the content of the three forms of PAs in durum wheat to different extents.