Automated effect-specific mammographic pattern measures

We investigate the possibility to develop methodologies for assessing effect specific structural changes of the breast tissue using a general statistical machine learning framework. We present an approach of obtaining objective mammographic pattern measures quantifying a specific biological effect, such as hormone replacement therapy (HRT). We compare results using this approach to using standard density measures. We show that the proposed method can quantify both age related effects and effects caused by HRT. Age effects are significantly detected by our method where standard methodologies fail. The separation of HRT subpopulations using our approach is comparable to the best methodology, which is interactive.      

An X-ray absorption approach to mixed and metallicity-sorted single-walled carbon nanotubes

Carbon based structures have been widely studied by X-ray absorption (XAS), also called NEXAFS, which is a very useful bulk probing method that allows examining the unoccupied density of states (DOS) and the site selective bonding environment. Two very well known spectral features in the XAS core level spectrum are the σ* and π* bands, and both have been analyzed in several studies for graphitic-like systems. However, among all the carbon materials, the unique one-dimensional electronic properties attributed to single-walled carbon nanotubes (SWCNTs) exhibit features that reveal clearly their electronic structure in the core level XAS spectrum. In this article, we outline the C1s response in XAS, which is related to the DOS of the conduction band in SWCNTs and its fine structure, revealed by experiments performed on metallicity-sorted SWCNT material. The progress in the identification of changes in the site selective conduction band electronic structure with XAS is discussed in detail.     

Genotyping of fig (Ficus carica L) via RAPD markers

The genetic diversity among 15 fig accessions (Ficus carica L), belonging to 9 renowned cultivars of the Calabrian fig collection, and 24 unidentified genotypes of figs also located in the Calabrian region of Southern Italy, was investigated by random amplified polymorphic DNA (RAPD) analysis. The genetic similarity values were calculated for the 39 samples, and a dendrogram was elaborated by cluster analysis according to the UPGMA algorithm. The generated DNA fragments grouped the samples into two main clusters of RAPD profiles. Most of the 24 unknown samples, coming from the Luzzi area, formed a unique cluster with high degree of genetic similarity. This indicates that it is possible to distinguish, at DNA level, the fig trees with an already well‐known potential to produce figs suitable for the drying process and possibly to specify cultivars with suitable features for industrial transformation. Copyright © 2005 Society of Chemical Industry     

Use of an Online Extraction Technique Coupled to Liquid Chromatography for Determination of Caffeine in Coffee,Tea, and Cocoa

Caffeine is the most widely studied psychoactive molecule in history due to its many pharmacological activities and a high number of biological and physiological effects. In literature, there is a great number of applications that describe extraction, identification, and quantification of caffeine in foods and beverages. For this purpose, an extraction step is followed by an analytical technique for the identification and quantification of caffeine. This work proposes an innovative method in which sample preparation, separation, and detection steps are unified in a single step. To the best of our knowledge, this is the first report on the determination of caffeine in coffee, tea, and cocoa by means of an online extraction coupled to a liquid chromatographic system equipped with a photodiode array detector. The developed methodology was validated in terms of sensitivity, detection limits, accuracy, and precision. The advantages of this technique are (i) a significant reduction of analysis time (more than 70%) and of solvents used (the extraction step is integrated in the chromatographic analysis), (ii) the whole procedure is thus completely automated drastically reducing possible operator errors to occur, and (iii) easily realized by using a conventional monodimensional liquid chromatography system.     

Monitoring Phenolic Composition of Maturing Maize Stover by High Performance Liquid Chromatography and Pyrolysis/Gas Chromatography/Mass Spectrometry*

Maize stovers collected every 14 days over an 84-day growth period were subjected to high-performance liquid chromatography with electrochemical detector (HPLC-ED) and pyrolysis/gas chromatography/mass spectrometry (PY/GC/MS) in order to monitor changes in the phenolic composition. Prior to HPLC-ED analyses, ground samples were sequentially extracted with (i) methanol, (ii) 0·1M sodium hydroxide and (iii) 2M sodium hydroxide in the presence of nitrobenzene to separate, respectively, free phenolic monomers, alkali-labile phenolic monomers and alkali-resistant lignin. In turn, solution (ii) was treated with alkaline nitrobenzene to obtain (iv) alkali-labile lignin. Pyrolysis was carried out on ground native samples by using a platinum heated filament pyrolyser. Increases in the absolute phenolic concentrations in the residues of 0·1M sodium hydroxide extraction and in the ratio of alkali-resistant lignin vs total lignin were observed by HPLC-ED during the first 28–42 days of maturation, reaching a steady level in the remaining maturation period. A linear increase of syringyl units vs guaiacyl units was for found the alkali-resistant lignin fraction over the entire period of maturation. Similar trends were showed by PY/GC/MS with regard to relative lignin content and syringyl/guaiacyl ratio. Both techniques showed their usefulness to gauge changes in the phenolic composition during the lignification process.     

Lactic acid bacteria ecology of three traditional fermented sausages produced in the North of Italy as determined by molecular methods

In this study the bacterial biodiversity during the maturation process of three traditional sausages produced in the North of Italy (Salame bergamasco, Salame cremonese and Salame mantovano) was investigated by using culture-dependent and -independent methods. Eleven plants, in the three provinces considered here, were selected because starter cultures were never used in the production. The bacterial ecology, as determined by plate counts, was dominated by lactic acid bacteria (LAB), with minor contribution of coagulase negative cocci and yeasts. After molecular identification of 486 LAB strains, the species more frequently isolated were Lactobacillus sakei and Lactobacillus curvatus. This evidence was also confirmed by PCR-Denaturing Gradient Gel Electrophoresis (DGGE). All the samples analyzed were characterized by the constant presence of L. sakei and L. curvatus bands. A richer biodiversity was only detected at the beginning of maturation. The results obtained by the molecular characterization of the L. sakei and L. curvatus and by the cluster analysis of the DGGE profiles highlighted a plant-specific population, rather than a geographic characterization of the products, underlining how the environmental and processing conditions are able to select specific microbiota responsible for the main transformations during the fermentation and ripening of the sausages.     

Determination of Toxic Elements in Nuts by Inductively Coupled Plasma Mass Spectrometry after Microwave-Induced Combustion

Microwave-induced combustion (MIC) of nuts in closed vessels was evaluated as a sample preparation method for further determination of As, Cd, and Pb by inductively coupled plasma mass spectrometry (ICP-MS) and Hg by flow injection cold vapor generation coupled to ICP-MS (FI-CVG–ICP-MS). Conventional microwave-assisted acid digestion (MW-AD) in pressurized vessels was also used for results comparison. Samples were wrapped in polyethylene films and combusted using 20 bar of oxygen and 50 μl of 6 mol l^?1 ammonium nitrate as aid for ignition. Sample masses up to 500 mg of hazelnuts, almonds, cashew nuts, Brazil nuts, and walnuts were combusted using 7 mol l^?1 HNO₃ as absorbing solution. Accuracy was evaluated using certified reference materials and an agreement better than 96% was obtained for all analytes. Using MIC, it was possible to obtain lower limits of detection (LODs) in comparison with those obtained by MW-AD. The LOD obtained using MIC was 3, 2, and 6 ng g^?1 for As, Cd, and Pb, respectively. The LOD for Hg by FI-CVG–ICP-MS was 7 ng g^?1. Residual carbon content obtained after decomposition by MW-AD and MIC was 20% and lower than 1.5%, respectively, showing the high efficiency of MIC and allowing the determination of toxic elements in samples with high fat content. With the use of MIC up to eight samples could be processed simultaneously and only diluted nitric acid was required minimizing physical interferences, reagent consumption, and waste generation.     

Validation of a 1-Day Analytical Diagnostic Real-Time PCR for the Detection of Salmonella in Different Food Meat Categories

Foodborne disease caused by Salmonella represents a worldwide public health problem. In Europe, salmonellosis is still the second most commonly recorded zoonosis. Since the standard culture method for detecting Salmonella (ISO 6579:2002) requires up to 5 days to produce results, the need to develop rapid methods represents an important issue for the authorities and the producers. The aim of the present study was the in-house validation, according to ISO 16140, of an open-formula diagnostic real-time PCR for the detection of Salmonella in all the different meat categories reported in the EU Regulations relative to microbiological criteria for food safety. The assay employed specific primers and a probe target within the ttrRSBCA locus, which allows the tetrathionate respiration in Salmonella. Selectivity, relative accuracy, relative sensitivity and relative specificity were established by testing 110 bacterial strains and 175 various edible meat samples. Results showed 100 % selectivity, 100 % relative accuracy, 100 % relative sensitivity and 100 % relative specificity of the real-time PCR when compared to the standard culture method used as reference. In addition, in order to minimize the effect of the competitive micro-flora naturally present on meat samples, a highly nutritious and selective commercial medium (ONE Broth Salmonella, Oxoid) was evaluated in comparison with the classical non-selective pre-enrichment broth (buffered peptone water). Results demonstrated that the ONE Broth Salmonella medium increases the growth of Salmonella in the presence of competitive micro-flora.