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

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

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