Quantification of Imatinib Plasma Levels in Patients with Chronic Myeloid Leukemia: Comparison Between HPLC–UV and LC–MS/MS

Measurement of imatinib plasma concentration can be useful to evaluate patient adherence to daily oral therapy, potential drug–drug interaction, treatment efficacy, and severe drug-related adverse events. The aim of this study was to correlate the two different blood level test methods, HPLC–UV and LC–MS/MS. We analyzed 162 plasma samples from patients treated with imatinib. We estimated the correlation between the two analytical methods on total data set and on five sets of patients grouped into different categories based on the drug-dose therapy. Finally, imatinib quantification was correlated with genetic data on the molecular response in monitoring follow-up of CML patients.     

Screening of plant-derived antifungal substances useful for the control of seedborne pathogens

The effect of plant essential oils, Solanum chilense and Aster sedifolius raw saponins, plant-derived colourants and Brassica carinata glucosinolate-containing tissues, for the control of seedborne fungi Alternaria dauci, Alternaria radicina, Colletotrichum lindemuthianum and Ascochyta rabiei, was evaluated. In vitro assays indicated that oregano essential oil, bark chestnut-derived colourant and B. carinata meals, showed a large spectrum of activity against all pathogens. S. chilense raw saponins were effective against A. dauci and A. radicina, while those from A. sedifolius also inhibited significantly C. lindemuthianum. Screening showed the antifungal potential of natural compounds.     

Detection of free-living and plankton-bound vibrios in coastal waters of the Adriatic Sea (Italy) and study of their pathogenicity-associated properties

Culturable vibrios were isolated from water and plankton fractions collected during an 18-month sampling study performed along the north-central coast of the Adriatic Sea (Italy). Unculturable Vibrio vulnificus and V. parahaemolyticus were detected in plankton fractions by polymerase chain reaction amplification of DNA sequences for cytotoxin-haemolysin and thermolabile haemolysin respectively. The presence of V. parahaemolyticus, V. vulnificus and V. cholerae virulence genes and the expression of pathogenicity-associated traits were analysed in all isolates. The results showed the spreading of these properties among the environmental isolates and confirm the need of both monitoring the presence of vibrios in coastal areas and studying their pathogenicity potential in order to properly protect human health.     

Chronic, sublethal nickel acclimation alters the diffusive properties of renal brush border membrane vesicles (BBMVs) prepared from the freshwater rainbow trout

Brush border membrane vesicles (BBMVs) were prepared from the kidneys of rainbow trout exposed acutely (72 h; 13,380 microg Ni L(-1)), chronically (11 months; 289 microg Ni L(-1)), or chronically and acutely, to waterborne nickel (Ni). Uptake of 63Ni into renal BBMVs was temperature-dependent and fitted a two component kinetic model composed of a saturable, Michaelis-Menten component prominent at lower Ni concentrations, and a moderate linear diffusive component apparent at higher Ni concentrations. Chronic Ni exposure reduced the permeability of the BBM to Ni, evidenced by a significantly reduced slope of the linear diffusive component of BBMV uptake. Efflux of Ni from 63Ni-loaded renal BBMVs was not significantly altered by acute Ni challenge. Both Ca2+ and Mg2+ inhibited uptake of Ni into renal BBMVs when present at a molar ratio to Ni of 1000:1. Mg2+-induced inhibition, however, was concentration-dependent and significant in BBMVs prepared from chronically Ni-acclimated fish at far lower molar ratios of 100 and 10 to 1. The data suggest that subtle, long-term modulation of membrane structure and function in the rainbow trout may be a compensatory response to chronic waterborne Ni exposure. Additionally, the data challenge the assumptions of constancy of the physiological parameters underlying physiologically based approaches to modeling metal toxicity. Such approaches are currently employed to derive water quality criteria for some metals.     

Inorganic mercury in the rock pool environment (Ligurian Sea)

In the present study the mercury concentration has been investigated in three components of a rock pool environment of Genoa Nervi (Ligurian Sea): water, particulate matter and Tigriopus fulvus (Harpacticoid copepod). The influence of some parameters (pH, temperature, salinity, rainfall) on mercury concentration has been evaluated. These data show that the mercury concentration is greater in Tigriopus males than in females and that the concentration also depends on temperature variations.     

Mesozooplankton assemblage and first record of Paracartia grani Sars G.O., 1904 (Copepoda: Calanoida) in the Western harbour of Genova (Ligurian Sea).

Harbours are characterized by high pollutant charge and by the occurrence of well adapted and resistant species. This paper reports the results of an annual plankton survey (May 1997-June 1998) carried out in the western harbour of Genova (Ligurian Sea) and in its mouth. Plankton samples were collected by horizontal trawls using a WP2 net. Copepods were the bulk of plankton in almost all samples. Eight copepod species were recognized: Paracalanus parvus and Acartia clausi were the most abundant. The first record of Paracartia grani in the harbour of Genova is here reported; this species, which is known to occur in polluted harbour waters of the Mediterranean Sea and was found in semi-confined Mediterranean and Atlantic coastal areas, was dominant during October 1997. Also Clausocalanus spp., Centropages typicus, Oithona helgolandica, Oithona nana, Farranula spp., Eurytemora spp., Isias clavipes and Lucicutia spp. were frequently sampled. Among other zooplankters, cladocerans, ostracods and tunicates occurred frequently, while cnidarians, mysids and chaetognaths showed low densities. These results show the occurrence of a well defined harbour plankton and point out the differences between harbour and neritic plankton of the Gulf of Genova.     

Rational Design of a Carrier Protein for the Production of Recombinant Toxic Peptides in Escherichia coli

Commercial uses of bioactive peptides require low cost, effective methods for their production. We developed a new carrier protein for high yield production of recombinant peptides in Escherichia coli very well suited for the production of toxic peptides like antimicrobial peptides. GKY20, a short antimicrobial peptide derived from the C-terminus of human thrombin, was fused to the C-terminus of Onconase, a small ribonuclease (104 amino acids), which efficiently drove the peptide into inclusion bodies with very high expression levels (about 200–250 mg/L). After purification of the fusion protein by immobilized metal ion affinity chromatography, peptide was obtained by chemical cleavage in diluted acetic acid of an acid labile Asp-Pro sequence with more than 95% efficiency. To improve peptide purification, Onconase was mutated to eliminate all acid labile sequences thus reducing the release of unwanted peptides during the acid cleavage. Mutations were chosen to preserve the differential solubility of Onconase as function of pH, which allows its selective precipitation at neutral pH after the cleavage. The improved carrier allowed the production of 15–18 mg of recombinant peptide per liter of culture with 96–98% purity without the need of further chromatographic steps after the acid cleavage. The antimicrobial activity of the recombinant peptide, with an additional proline at the N-terminus, was tested on Gram-negative and Gram-positive strains and was found to be identical to that measured for synthetic GKY20. This finding suggests that N-terminal proline residue does not change the antimicrobial properties of recombinant (P)GKY20. The improved carrier, which does not contain cysteine and methionine residues, Asp-Pro and Asn-Gly sequences, is well suited for the production of peptides using any of the most popular chemical cleavage methods.     

zucchini and squash encode two putative nucleases required for rasiRNA production in the Drosophila germline

RNAi is a widespread mechanism by which organisms regulate gene expression and defend their genomes against viruses and transposable elements. Here we report the identification of Drosophila zucchini (zuc) and squash (squ), which function in germline RNAi processes. Zuc and Squ contain domains with homologies to nucleases. Mutant females are sterile and show dorsoventral patterning defects during oogenesis. In addition, Oskar protein is ectopically expressed in early oocytes, where it is normally silenced by RNAi mechanisms. Zuc and Squ localize to the perinuclear nuage and interact with Aubergine, a PIWI class protein. Mutations in zuc and squ induce the upregulation of Het-A and Tart, two telomere-specific transposable elements, and the expression of Stellate protein in the Drosophila germline. We show that these defects are due to the inability of zuc and squ mutants to produce repeat-associated small interfering RNAs.