Solid-phase microextraction gas chromatography/mass spectrometry: a new method for species identification of truffles

This study describes a rapid method to identify different truffle species by analysis of their volatile compound fraction using static headspace solid-phase microextraction gas chromatography/mass spectrometry. The volatile organic compounds (VOCs) were extracted using a new 2-cm 50/30 microm DVB/CAR/PDMS fiber placed for 10 min in the headspace of the truffle sample with the vial maintained at 20 degrees C (in a thermostatically controlled analysis room). The mass spectra of the VOC chromatograms were represented as 'fingerprints' of the analysed samples. Next, stepwise factorial discriminant analysis afforded a limited number of characteristic fragment ions that allowed a classification of the truffle species studied. This new method provides an effective approach to rapid quality control and identification of truffle species by analysis of their volatile fraction. Moreover, this method offers the advantage of minimizing thermal, mechanical, and chemical modifications of the truffles, thereby reducing the risk of analytical artifacts.     

Characterization of the volatile organic compounds of Italian 'Fossa' cheese by solid-phase microextraction gas chromatography/mass spectrometry

Fossa cheese is an Italian hard cheese, ripened for up to 3 months in underground pits dug into tuffaceous rock. During this period, the cheese develops a unique flavour and intense and somewhat piquant aroma. Solid-phase microextraction gas chromatography/mass spectrometry (SPME-GC/MS) was utilized to characterize the volatile organic compounds (VOCs) of Fossa cheese. A total of 75 VOCs were separated and identified; in particular, the major class of compounds found in the cheeses ripened in the pits were the esters of fatty acids. Discriminant analysis of volatile profiles allowed us to distinguish between cheeses in different stages of seasoning (60-day-old cheese and cheese ripened an additional 90 days in and out of the pits).     

Extraction of the x-dependence of the non-perturbative QCD b-quark fragmentation distribution component

Using recent measurements of the b-quark fragmentation distribution obtained in events, registered at the Z pole, the non-perturbative QCD component of the distribution has been extracted independently of any hadronic physics modelling. This distribution depends only on the way the perturbative QCD component has been denned. When the perturbative QCD component is taken from a parton shower Monte Carlo, the non-perturbative QCD component is rather similar with those obtained from the Lund or Bowler models. When the perturbative QCD component is the result of an analytic NLL computation, the non-perturbative QCD component has to be extended in a non-physical region and thus cannot be described by any hadronic modelling. In the two examples, used to characterize these two situations, which are studied at present, it happens that the extracted non-perturbative QCD distribution has the same shape, being simply translated to higher-x values in the second approach, illustrating the ability of the analytic perturbative QCD approach to account for softer gluon radiation than with a parton shower generator.     

Constraints on new physics from the quark mixing unitarity triangle

The status of the unitary triangle beyond the standard model including the most recent results on Deltam[s] on dilepton asymmetries and on width differences is presented. Even allowing for general new physics loop contributions the unitarity triangle must be very close to the standard model result. With the new measurements from the Fermilab Tevatron, we obtain for the first time a significant constraint on new physics in the Bs sector. We present the allowed ranges of new physics contributions to DeltaF=2 processes and of the time-dependent CP asymmetry in Bs-->J/psivarphi decays.     

Headspace solid-phase microextraction with gas chromatography and mass spectrometry in the investigation of volatile organic compounds in an ectomycorrhizae synthesis system

Ectomycorrhizae formation represents one of the most significant steps in the truffle life cycle and is determined by a complex molecular signaling between two symbionts. In order to understand the molecular pathway of ectomycorrhiza development, we focused on the signaling interaction between the ectomycorrhizal fungus Tuber borchii Vittad. and the Tilia americana L. plant roots. The medium of a pre-symbiotic (T. americana-T. borchii) in vitro system was analysed by headspace solid-phase microextraction coupled with gas chromatography and mass spectrometry. In total, 73 volatile organic compounds (VOCs) were identified. Twenty-nine of these VOCs were produced only during the interaction phase between the two partners, leading to a hypothesis that these molecules might act as molecular messengers in order to pilot the ectomycorrhizae formation.     

Solid-phase microextraction gas chromatography/ion trap mass spectrometry and multistage mass spectrometry experiments in the characterization of germacrene D

Germacrene D is a vegetable pheromone utilized in interactions among organisms belonging to different species. For the first time, using solid-phase microextraction/gas chromatography/ion trap mass spectrometry, the presence of this compound was detected in an in vitro mycorrhizal synthesis system where the mycelium of the ectomycorrhizal fungus Tuber borchii Vittad. interacts with the plant Tilia Americana L. From this symbiosis, a new structure, called ectomycorrhiza, is formed where the two symbionts exchange nutrients and metabolites. It seems that only after this interaction can the mycelium develop the fruitbody, commonly known as truffle. The results obtained allowed us to ascertain that germacrene D was synthesized by the plant exclusively in the presence of T. borchii. The originality of these data prompted us to hypothesize that this compound could be involved in the first step of ectomycorrhiza formation, as it is able to stimulate specific fungi receptors. In fact, plants release hundreds of secondary metabolites that are important in their interactions with other organisms.     

Determination of microbial volatile organic compounds from Staphylococcus pasteuri against Tuber borchii using solid-phase microextraction and gas chromatography/ion trap mass spectrometry

The mycelium of Tuber borchii Vittad., a commercial truffle species, is used as a model system for in vitro ectomycorrhizal synthesis, infected seedling production and biotechnological applications. Our fungal cultures were accidentally contaminated with a Staphylococcus pasteuri strain, showing a strong antifungal activity against T. borchii mycelium. In order to identify the antifungal volatile agents produced by S. pasteuri, solid-phase microextraction (SPME) with gas chromatography and mass spectrometry (GC/MS) was used. Using this method 65 microbial volatile organic compounds (MVOCs), synthesized by this bacterium in either single or in fungal-bacterial dual culture, were identified. SPME combined with GC/MS may be a useful method for the determination of MVOCs involved in the antifungal activity. These results showed that bacteria with unusual biological activities could be a major problem during large-scale production of inoculum for truffle-infected seedling.     

A fluorescence-based sequence-specific primer PCR for the screening of HLA-B(*)57:01

Abacavir (ABC) is an antiretroviral drug highly effective in the treatment of HIV, but its intake can cause severe hypersensitivity reaction (HSR). A strong association between HLA-B(*)57:01 and ABC HSRs was reported by several studies, which demonstrated that HLA-B(*)57:01 screening had a 100% negative predictive value and that it could accurately identify patients at high risk of ABC HSRs. We propose a new sequence-specific primer PCR assay based on fluorescence detection through CE which is highly sensitive, allowing the use of non-infective sources of DNA such as saliva and buccal swabs, in addition to blood and reproducible, allowing automation of the analytical process. The results of our study were first compared with a standard sequence-specific primer PCR technique and reported a concordance of 100%, and then a blind external validation further confirmed the accuracy of our method.     

Studies on the use of Sepharose-N-Aminohexanoyl-Glucosamine for purification of rabbit red blood cell hexokinase

N-acetyl glucosamine is a competitive inhibitor (K _t= 0.7 mM) of red blood cell hexokinase with respect to glucose. This property has been utilized for the purification of hexokinase by means of Sepharose-TV-aminohexanoyl-glucosamine. Studies with this matrix have proved that ionic strength and pH play a very important role in the binding of hexokinase to the affinity column. Therefore their control is essential in order to minimize nonspecific binding and to maximize the purification. Methods for rejuvenation of columns, the effect of protein concentration, and the nature of the binding are also discussed in this paper.