9 research outputs found
Integrated analytical approach in veal calvesadministered the anabolic androgenic steroidsboldenone and boldione: urine and plasma kineticprofile and changes in plasma protein expression
Surveillance of illegal use of steroids hormones in cattle breeding is a key issue to preserve human
health. To this purpose, an integrated approach has been developed for the analysis of plasma and
urine from calves treated orally with a single dose of a combination of the androgenic steroids boldenone
and boldione. A quantitative estimation of steroid hormones was obtained by LC-APCI-QMS/
MS analysis of plasma and urine samples obtained at various times up to 36 and 24 h after
treatment, respectively. These experiments demonstrated that boldione was never found, while
boldenone a- and b-epimers were detected in plasma and urine only within 2 and 24 h after drug
administration, respectively. Parallel proteomic analysis of plasma samples was obtained by combined
2-DE,MALDI-TOF-MS and mLC-ESI-IT-MS/MS procedures. A specific protein, poorly represented
in normal plasmasamples collected before treatment,was found upregulated even 36 h after
hormone treatment.Extensivemassmapping experiments proved this component as an N-terminal
truncated form of apolipoprotein A1 (ApoA1), a protein involved in cholesterol transport. The
expression profile of ApoA1 analysed byWestern blot analysis confirmed a significant and time dependent
increase of thisApoA1 fragment. Then, provided that further experiments performed with
a growth-promoting schedule will confirm these preliminary findings, truncated ApoA1 may be
proposed as a candidate biomarker for steroid boldenone and possibly other anabolic androgens
misuse in cattle veal calves, when no traces of hormones are detectable in plasma or urine
Late apoptotic effects of taxanes on K562 erythroleukemia cells: apoptosis is delayed upstream of caspase-3 activation.
The efficacy of taxanes on human leukemia cells is the object of intensive in vitro investigation concerning the influence of cell-type-specific characteristics on cytotoxic response to drugs. The present study dissects the response to taxanes of HL60 acute myelomonocytic leukemia and of K562 chronic myelogenous leukemia, in parallel over a 72-hr time-span. The kinetics of cytotoxicity following pulsed and continuous exposure to either taxol or taxotere showed a delayed response of K562 cells independently of dose and type of exposure. In K562 cells, apoptosis became evident at 48 hr and prominent at 72 hr of treatment. These events were mirrored by delayed kinetics of caspase-3 activation. Comparable microtubule targeting was demonstrated in HL60 and in K562 cell lines, as bcl-2 and raf-1 were phosphorylated following treatment with taxanes. These observations indicate that early activation processes were responsible for apoptosis, but that the delay was determined by other factors. In addition, cell-free-system experiments excluded the presence of excess nuclear and/or cytoplasmic inhibitory factors and demonstrated that K562 cells possess a fully competent caspase system which can be readily activated. Processing of caspase-3 pro-enzyme was in fact increased by addition of cytochrome c. These results extend to taxol and taxotere the notion that drug-induced apoptosis is delayed upstream of caspase-3 activation in K562 cells, that such kinetics is independent of drug concentration and exposure time, and that it is linked to intrinsic cellular characteristics mapping between bcl-2 phosphorylation and cytochrome c release
Characterisation of table olive cultivar by NIR spectroscopy
NIR spectroscopy is proposed as a rapid and effective analytical method for the identification of the cultivar of table olives. The study was particularly focused on the Taggiasca cultivar. A set of 46 samples representative of the Taggiasca production area was analysed together with 43 samples of table olives of different cultivars. After feature selection, LDA and SIMCA were applied to the NIR data as classification and class-modelling techniques, respectively. The excellent results obtained by NIR spectroscopy (mean specificity of the models 82.0%, sensibility >90%) were then compared to those obtained by the gas chromatographic (GC) determination of the fatty acids composition of the oils extracted from the samples of table olives. Finally, in order to test the potential synergy among different sources of information, the employed NIR and chemical variables were joined, but only a small improvement of the results obtained by NIR alone was reache
A hemolytic peptide from the mycophilic fungus Sepedonium chrysospermum (Bull.) Fr.
none6SANGUINETI E; COSULICH E; SALIS A; DAMONTE G; M. MARIOTTI; ZOTTI MSanguineti, Elisa; Cosulich, E; Salis, Annalisa; Damonte, Gianluca; Mariotti, Mauro; Zotti, Mirc
Steric Factors Controlling the Surface Hybridization of PCR Amplified Sequences
This study elucidated the hybridization behavior of surfacebound
oligonucleotides to their longer PCR-amplified
targets. The screen-printed gold surface of disposable
electrodes was the platform onto which thiol-tethered
oligonucleotides (21-mer) were immobilized by chemisorption.
As a model case, 600-bp amplicons were
studied. Surface hybridization was monitored by means
of an enzyme-linked assay with electrochemical detection.
Use of different surface-tethered probe sequences over a
wide range of surface densities was explored to achieve
the highest duplex yield. Both the surface coverage by the
probe and its relative position on the target strand were
found to control the efficiency of capture of the target
sequence. Interfacial hybridization occurred with the
highest efficiency for a probe coverage of 2.9 1012
molecules/cm2 and when the 3¢ end of the amplicon was
involved. An unusual (bell-shaped) response/amplicon
concentration profile was additionally found. It was hypothesised
that when the amount of solution-phase target
is relatively high, random collisions make reannealing of
the 600-bp strands favored over formation of the surfacetethered
probe-amplicon complex. This paper also describes
a strategy to enhance the sensitivity of enzymelinked
hybridization assays. Such a strategy relies on
formation, around the long target sequence, of dendriticlike
structures, which could offer multiple anchoring
points for the enzyme conjugate. The results shown in this
work might have great significance for the practical
application of hybridization to oligonucleotide chips
Interdisciplinary study for the evaluation of biochemicalalterations on mussel Mytilus galloprovincialis exposedto a tributyltin-polluted area
An interdisciplinary approach was employed to
monitor the concentration and the effects of butyltin compounds
in mussels (Mytilus galloprovincialis). Tissues from
animals exposed to a marine area (Vado Ligure harbour)
with a high concentration of tributyltin (TBT) were analysed
and compared with control samples. TBT concentrations
were measured by gas chromatography–mass spectrometry
and the protein pattern in gill tissues was studied by proteomic
analysis. Several proteomic signatures associated with
contaminant exposure were observed; spots that were significantly
increased in all contaminated samples were
identified by mass spectrometry as fragments of β-tubulin.
The degradation of β-tubulin was then confirmed by western
blot analysis with specific anti-β-tubulin antibody. The effects
observed on mussel gills after exposure in the TBT-polluted
area are discussed