Degradation of aromatic hydrocarbons by white-rot fungi in a historically contaminated soil

Phanerochaete chrysosporium NRRL 6361 and Pleurotus pulmonarius CBS 664.97 were tested for their ability to grow under nonsterile conditions and to degrade various aromatic hydrocarbons in an aged contaminated soil that also contained high concentrations of heavy metals. After 24 days fungal incubation, carbon-CO2 liberated, an indicator of microbial activity, reached a plateau. At the end of the incubation time (30 days), fungal colonization was clearly visible and was confirmed by ergosterol and cell organic carbon determinations. In spite of unfavorable pH (around 7.4) and the presence of heavy metals, both fungi produced Mn-peroxidase activity. In contrast, laccase and aryl-alcohol oxidase were detected only in the soil treated with P. pulmonarius CBS 664.97 and lignin-peroxidase in that with P. chrysosporium NRRL 6361. No lignin-modifying enzyme activities were present in non-inoculated soil incubated for 30 days (control microcosm). Regardless of the fungus employed, a total removal of naphtalene, tetrachlorobenzene, and dichloroaniline isomers, diphenylether and N-phenyl-1-naphtalenamine, was observed. Significant release of chloride ions was also observed in fungal-treated soil, in comparison with that recorded in the control microcosm. Both fungi led to a significant decrease in soil toxicity, as assessed using two different soil contact assays, including the Lepidium sativum L. germination test and the Collembola mortality test.     

Enigmatic dinosaur trackways from the lower jurassic (Pliensbachian) of the Sarca valley,Northeast Italy

Four large limestone blocks with dinosaur footprints have been discovered in a large landslide situated in the Sarca Valley (northeast Italy). The trampled sediments have been ascribed to the upper part (Pliensbachian) of the Lower Jurassic Calcari Grigi Formation, which in the Sarca Valley consists of oolitic‐bioclastic limestones. These carbonates were originally deposited on the western margin of the Trento Platform, a paleogeographic structural high that was located in the northern tropical belt at the westernmost end of Tethys. Several footprints form trackways of quadrupedal dinosaurs. The data suggest the presence, within the ichnoassociation, of possible prosauropods and basal Thyrephora. This inference is consistent with the chronological and geographical distribution of both prosauropods and Thyreophora.     

Cellular permeation pathways in a leaky epithelium: the human amniochorion

The presence of cellular permeation pathways in human fetal membranes at term was evaluated. Electrical parameters (transepithelial potential [TEP], and conductance [Gt], and intracellular potentials [cell PD]), and water and urea diffusional coefficients (Pdw, Pdu), were determined in Ussing-like chambers. In amniochorion, the TEP was practically 0 (0.1 +/- 0.03 mV), and the Gt very high (144 +/- 14 mS/cm2). The Cell PD of amnion cells was -37 +/- 3 mV. Increasing the [K+] of the amniotic perfusate between 5.8 and 125.8 mM depolarized the cells with a slope of 23 mV. The deletion of Na+ hyperpolarized the cells, whereas amiloride and ouabain depolarized them. The Pdw and Pdu were determined in intact amnion and chorion and in their epithelial cell layers. The Pdw/Pdu ratio in amnion was 4.0, and 7.0 in its cell layer; the ratio in chorion was 2.5, and 3.3 in its cell layer. The amniochorion is a leaky structure, but its cellular layers possess definite transcellular permeation pathways. The ionic conductances in amnion cells are complex, with the Cell PD being determined by at least K+ and Na+ conductances, and ouabain- and amiloride-sensitive pathways. The amnion is a more effective diffusional barrier to water and urea than chorion is; its diffusional characteristics are comparable to those of nystatin-treated lecithin: cholesterol bilayers and the membranes of human erythrocytes.     

Fermentation of xylo-oligosaccharides by Bifidobacterium adolescentis DSMZ 18350: kinetics, metabolism, and β-xylosidase activities

Xylo-oligosaccharides (XOS) are sugar oligomers of β-1,4-linked xylopyranosyl moieties which exert bifidogenic effect and are increasingly used as prebiotics. The kinetics and the metabolism of Bifidobacterium adolescentis DSMZ 18350 growing on XOS and xylose were investigated. The growth rate was higher on XOS, but greater biomass yield was attained on xylose. Unlike other prebiotics, XOS oligomers were utilized simultaneously, regardless of their chain length. Throughout XOS utilization, xylose concentration slightly increased, being not neatly consumed and remaining unfermented. During growth on XOS, β-xylosidase activity was present in the cytosol, but it occurred in the supernatant as well. A β-1,4-xylolytic enzyme was purified from the supernatant of XOS cultures. The enzyme, a homotetramer of a 39-kDa single protein, was capable of complete XOS hydrolysis and exhibited maximum activity at pH?6.0 and 55 °C. Based on the molecular weight, the protein can be ascribable to the product of the gene BAD_1527, the activity of which has been inferred as an endo-β-1,4-xylanase, but has not been characterized so far. This β-1,4-xylolytic enzyme, found to be active in the cultural supernatant, gives a reason for the never explained accumulation of the monosaccharides in the media of bifidobacterial cultures growing on XOS, without excluding the major role of the intracellular hydrolysis of the imported oligomers.     

Role of percutaneous pelvic node aspiration cytology in the management of bladder carcinoma

Six of the 23 patients with bladder carcinoma who underwent percutaneous pelvic node aspiration biopsy cytology for staging purposes showed positive findings from nodal chains. Pelvic lymphadenectomy revealed no false-positive or false-negative cytologic results. The fine needle aspiration of opacified pelvic nodes under fluoroscopic control is of great diagnostic value, with a high accuracy in detecting nodal extension of bladder carcinoma and providing a rational basis for proper therapy. Positive aspiration results may be accepted as a basis for therapeutic decisions. Since the metastatic involvement of multiple nodes makes any therapeutic treatment as well as cystectomy completely useless, positive aspirations from more than two nodes may spare patients an unnecessary radical surgery.     

Application of Fluorescence Emission for Characterization of Albendazole and Ricobendazole Micellar Systems: Elucidation of the Molecular Mechanism of Drug Solubilization Process

Albendazole (ABZ) and ricobendazole (RBZ) are referred to as class II compounds in the Biopharmaceutical Classification System. These drugs exhibit poor solubility, which profoundly affects their oral bioavailability. Micellar systems are excellent pharmaceutical tools to enhance solubilization and absorption of poorly soluble compounds. Polysorbate 80 (P80), poloxamer 407 (P407), sodium cholate (Na-C), and sodium deoxycholate (Na-DC) have been selected as surfactants to study the solubilization process of these drugs. Fluorescence emission was applied in order to obtain surfactant/fluorophore (S/F) ratio, critical micellar concentration, protection efficiency of micelles, and thermodynamic parameters. Systems were characterized by their size and zeta potential. A blue shift from 350 to 345 nm was observed when ABZ was included in P80, Na-DC, and Na-C micelles, while RBZ showed a slight change in the fluorescence band. P80 showed a significant solubilization capacity: S/F values were 688 for ABZ at pH 4 and 656 for RBZ at pH 6. Additionally, P80 micellar systems presented the smallest size (10 nm) and their size was not affected by pH change. S/F ratio for bile salts was tenfold higher than for the other surfactants. Quenching plots were linear and their constant values (2.17/M for ABZ and 2.29/M for RBZ) decreased with the addition of the surfactants, indicating a protective effect of the micelles. Na-DC showed better protective efficacy for ABZ and RBZ than the other surfactants (constant values 0.54 and 1.57/M, respectively), showing the drug inclusion into the micelles. Entropic parameters were negative in agreement with micelle formation.     

The challenge for identifying the fungi living inside mushrooms: the case of truffle inhabiting mycelia

The Tuber ascomata (truffles) are a microhabitat for bacteria, viruses, and fungi (yeasts and filamentous fungi). In this survey, we tried to develop a method that would make it possible to define the mycobiome of the truffle-inhabiting filamentous fungi using culture independent molecular methods. The nested quantitative Real-Time PCR allowed us to demonstrate that each truffle is home to multiple species of filamentous fungi and that their DNA is present within the healthy ascoma at the ratio of 10^?6 compared to that of the truffle. Probably due to their insignificant presence, Denaturing Gradient Gel Electrophoresis of the amplification of ITS amplicons showed only those of the host. Based on the results, the possibilities of being able to detect the fungicolous fungi present in very small amounts within a fungal host are discussed.     

Bacillus thuringiensis CrylAa δ-Endotoxin Affects the K+/Amino Acid Symport in Bombyx mori Larval Midgut

The mechanical properties of brush border membrane vesicles, BBMV, from rabbit kidney proximal tubule cells, were studied by measuring the initial and final equilibrium volumes of vesicles subjected to different osmotic shocks, using cellobiose as the impermeant solute in the preparation buffer. An elevated intracellular hydrostatic pressure was inferred from osmotic balance requirements in dilute solutions. For vesicles prepared in 18 and 85 mosm solutions, these pressures are close to 17 mosm (290 mm Hg). The corresponding membrane surface tension is 6.0 × 10⁻⁵ N cm⁻¹ while the membrane surface area is expanded by at least 2.2%. When these vesicles are exposed to very dilute solutions the internal hydrostatic pressure rises to an estimated 84 mosm (1444 mm Hg) just prior to lysis. The corresponding maximal surface tension (pre-lysis) is 18.7 × 10⁻⁵ N cm⁻¹, and the maximal expansion of membrane area is 6.8%. The calculated area compressibility elastic modulus was 2.8 × 10⁻³ N cm⁻¹. Received: 8 August 1996/Revised: 4 March 1997     

Association of protein amount polymorphism (PAP) among maize lines with performances of their hybrids

Summary It has been suggested that molecular foundations of phenotypic diversity reside in the variability of genome expression. This variability can be appraised through the polymorphism of individual protein amounts (PAP: protein amount polymorphism). Eight maize inbred lines and ten of their single-cross hybrids were analyzed by two-dimensional polyacrylamide gel electrophoresis in order to examine the potential of PAP for predicting hybrid vigor. The 28 possible pairs of lines were characterized for: (i) the number, H of expected heterozygous structural loci in their hybrid, in the sample of loci revealed by 2D-PAGE; (ii) four distance indices based on PAP; (iii) the hybrid values for five agromorphological characters measured in four different year/locations. For the subset of ten hybrids analyzed by 2D-PAGE, the number of cases of nonadditive inheritance (NA) was also counted. Whereas H appeared to be related neither to the PAP indices, nor to NA, nor to hybrid performances, PAP indices were correlated to NA, and both were positively associated to hybrid performances. The possibility that PAP is responsible for quantitative trait variation is discussed. This could result in the definition of biological predictors of heterosis.     

Amino acid uptake in plasma membrane vesicles isolated from proliferating tumor cells and tissues

Summary The transport of L-alanine, a natural substrate of system A, across plasma membrane vesicle preparations has been studied in the early stages of rat DENA-PH hepato-carcinogenesis and in a very undifferentiated rat ascites hepatoma cell line (Yoshida AH-130) in the exponential and stationary phase of growth.Kinetic analyses indicated an increase of the V_max value in DENA-PH-treated rats 30 h after partial hepatectomy as well as in exponential growing Yoshida ascites cells. In DENA-PH-treated rats the Km value was drastically reduced 7 and 60 days after surgery, when enzyme-altered hyperplastic and preneoplastic lesions were present in rat liver. Drastically reduced Km values were also found in Yoshida ascites cells.The results suggest that an altered alanine transporter might take place in liver plasma membranes from carcinogen-treated rats. This appears to occur also in an established tumor cell line, grown in vivo.Abbreviations AAF 2-acetylaminofluorene - DENA diethylnitrosamine - PH partial hepatectomy - PMSF phenylmethanesulfonyl fluoride