Detection and activity evaluation of radical scavenging compounds by using DPPH free radical and on-line HPLC-DPPH methods

The antiradical activity of crude extracts (80% methanol, 20% water) of S. officinalis, S. glutinosa, S. sclarea and S. aethiopis was determined using the 2,2-diphenyl-1-picrylhydrazyl (DPPH·) radical scavenging method. For validation of this method several well known antioxidants (ascorbic acid-6-palmitate, gallic, chlorogenic, ferulic, caffeic, uric, gentisic and vanillic acids, catechin, quercetin, epicatechin, phloridzin, rutin and naringin) were investigated additionally. In these experiments ascorbic acid-6-palmitate had highest antiradical activity. Within the group of phenolic acids gentisic acid had the highest antiradical activity comparing with the other tested phenolic acids. Uric acid, vanillic acid, phloridzin and naringin have a much lower antiradical activity. Different reaction kinetics behaviour was observed. The validated DPPH radical scavenging method was applied to the evaluation of the antiradical activity of plant extracts. The Salvia extracts showed very high antiradical activity towards the DPPH·. An on-line HPLC-DPPH method was developed using a methanolic solution of DPPH· for a rapid detection of radical scavenging components after HPLC separation. The HPLC-DPPH on-line method was also applied to a screening of several radical scavenging components in plant extracts as well as for quantitative analysis. The HPLC analysis revealed the presence of several radical scavenging components in the all Salvia extracts. This HPLC-DPPH on-line method can also be used for quantitative determination of radical scavenging compounds.     

Chemometric studies of some trace elements in cows' milk

Summary The matrix of the correlation between the concentrations of 11 trace elements determined in 45 samples of cows' milk was used as a starting matrix for principal component analysis. The space dimension was reduced from 11 variables to 5 principal components, accounting for about 80% of the total variance. After an orthogonal rotation, the first factor (F1) was found to be positively correlated with Cr, Mn, and Fe, F2 positively correlated with Ni and negatively with Cu, F3 positively with Cd and Pb, F4 positively with Zn, Sr, and Mo and F5 positively correlated with Al and Sr. Element clusters appear to be determined by their origin. Some clusters are confirmed by principal factor analysis. A plot of the principal component scores was also applied to the differentiation of Italian Milk from different geographical origins.

---

Chemometrische Untersuchungen über einige Spurenelemente in pasteurisierter Kuhmilch

Zusammenfassung Die Analyse der Hauptkomponenten wurde auf die Korrelationsmatrix zwischen Konzentrationen von 11 Spurenelementen in 45 Kuhmilchproben angewandt. Die Raumdimension von 11 Variablen wurde auf 5 Hauptkomponenten die 80% der totalen Varianz ausmachen, reduziert. Die orthogonal-rotierte Faktorenmatrix zeigt, daß Cr, Mn und Fe an die erste Hauptkomponente gebunden sind, Ni und Cu an die zweite, Cd und Pb an die dritte, Zn, Sr und Mo an die vierte and Al and Sr an die fünfte. Die Herkunft der Gruppen von Spurenelementen werden diskutiert. Einige Gruppen wurden bei der Analyse der Hauptfaktoren bestatigt. Das Diagramm der Hauptkomponentenwerte wurde auf die Unterscheidung von Milch aus zwei geographischen Herkünften angewandt.

     

Environmental exposure to asbestos and other inorganic fibres using animal lung model

Professional exposure to asbestos fibres is widely recognized as very dangerous to human health and for this reason many countries have banned their commercial uses. People, nevertheless, continue to be exposed to low dose of asbestos from natural and anthropogenic sources still in loco, for which the potential hazard is unknown.The aim of this research is to assess environmental exposure in an area with outcropping serpentinite rocks, which bear asbestos mineralizations, using sentinel animals which are a non-experimental animal model. We studied the burden of inorganic fibres in cattle lungs which come from two areas in Italy's Western Alps bearing serpentinitic outcrops: Susa Valley with a heavy anthropization and Lanzo Valleys, with a minor human impact. The identification and quantification of inorganic fibres were performed by Scanning Electron Microscope (SEM) and Energy Dispersive Spectrometer (EDS). In comparison to humans, studies of animals have some advantages, such as no occupational exposure or history of smoking and, in the case of cattle, a sedentary life restricted to one region.Results spotlight that over than 35% of inorganic fibres found both in Susa and Lanzo valleys, belong to asbestos mineralogical species (asbestos tremolite/actinolite, chrysotile s.s., asbestos grunerite, crocidolite). We also observed a higher concentration of artificial fibrous products in Susa samples showing a correlation with the level of anthropization.These results confirm that sentinel animals are an excellent model to assess breathable environmental background because it is possible to eliminate some variables, such as unknown occupational exposure.     

Enumeration of protozoan ciliates in activated sludge: Determination of replicate number using probability

A new approach to the enumeration of ciliate protozoa in activated sludge is described. A 25 μL sub-sample volume is optimal for routine analysis using a standard slide and 24 × 24 mm cover slip requiring between 20 and 40 min per sub-sample for full enumeration and identification of species. However, to achieve high probability (≥95%) of recovering all species large numbers of replicates are required (i.e. 23-47). To achieve high probabilities of recovery using less replicates it is necessary to neglect rare species with low densities (<0.5%); based on the assumption that they do not play a significant role in plant performance. The precise number of replicates required for different probabilities of recovering species is determined by conducting an initial pilot survey analysing a minimum of 8 replicates and using a probability equation to determine the optimum replicate number for that particular plant. Six replicate 25 μL sub-samples provided excellent species recovery (90-95% excluding up to 3 rare species), while analysing just two or three replicates, as commonly used in previous wastewater studies, only gave probabilities of 25 and 50% respectively for the same recovery. Ciliate analysis should be completed within 8 h of collection with significant changes in community structure occurring beyond this period.     

Cadmium in roadside grapes

The concentration of cadmium in unwashed wine grapes and leaves is considered as a function of the distance from the suspected source, an isolated straight road. Cadmium levels were determined by a.a.s. with electrothermal atomisation in a graphite furnace. The grape and leaf samples display an asymptotic decrease in cadmium concentration (c) according to the distance (d) from the edge of the road. The observed data can be fitted with the exponential function c=A+B exp (-Cd), the parameters being estimated by least-squares non-linear regression. The simple model allows the determination of the cadmium level at d→∞ (A) and at d=0 (A+B). The results obtained indicate that the effect of the presence of the road is not detectable beyond a distance of 50 m.     

Donkeys’ milk protein fraction investigated by electrophoretic methods and mass spectrometric analysis

Donkeys' milk has attracted a growing interest in human nutrition, particularly in the dietotherapy of patients affected by cows' milk protein allergy. In this paper, donkeys' milk protein fractions were analysed by isoelectric focusing (IEF) and sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). Four different IEF patterns were observed out of 51 individual milk samples. Thirty-six samples presented a common pattern, used as a reference, whereas 15 were characterized by the presence or absence of protein bands. A single sample, lacking two bands of the casein fraction, showed an apparent reduced amount of caseins, by SDS-PAGE as well as by chromatographic analysis. Mass spectrometry and high-performance liquid chromatography were used to characterize the IEF patterns. Compared with the reference, one pattern lacked two α_s1-casein bands, another lacked two β-lactoglobulin II bands and one accounted for a different β-lactoglobulin II variant. No signals related to putative α_s2- and κ-casein were observed.     

Lead in grapes exposed to automobile exhaust gases

Lead accumulates in grapes exposed to automobile exhaust gases, the concentration of the metal diminishing parabolically with the logarithm of the distance from the source of pollution. The maximum extrapolated figure was 4 to 6 ug per g of dried grapes, or about 10 times the normal content. The pollution falls to zero 100 to 120 m away from the roadway.     

Multivariate data analysis of sea waters and mussels in relation to pollution sources of trace elements

Summary The total concentration of Cu, Zn and Pb in surface sea waters from the Bay of Muggia (Gulf of Trieste, Northern Adriatic Sea) was determined by anodic stripping voltammetry. The association of these trace elements in relation to the known sources of pollution was discussed. The content of eight trace elements (Mn, Co, Ni, Cu, Zn, Cd, Hg, Pb) in the soft part of mussels (Mytilus galloprovincialis Lamarck) is also considered. The wild molluscs were sampled in the harbour of Trieste, in the proximity of an important city sewer. Principal component analysis was used to analyse the correlation matrix obtained from an 8 × 43 data matrix after a logarithmic transformation of the concentration variables. Eight variables were reduced to four principal components, which explained 80% of the total variance. The orthogonally rotated factor matrix shows that Co, Ni, Cd, and Pb are associated with the first principal component, Cu and Zn to the second, Hg to the third and Mn to the fourth principal component. The results of this multi-variate data analysis are compared with those already obtained from two sampling sites in the Bay of Muggia and the origin of some trace metals in the soft part of mussels from the Gulf of Trieste is discussed.

---

Mehrdimensionale Analyse der Daten von Seewasser und Muscheln in Beziehung zu Verunreinigungen mit Spurenelementen

Zusammenfassung Der Gesamtgehalt von Cu, Zn, Pb in den oberen Schichten des Seewassers aus der Bucht von Muggia (Golf von Triest) wurde nach einer puls-inversvoltammetrischen Methode untersucht. Es wurden die Beziehungen dieser Spurenelemente zu den bekannten Verunreinigungsursachen diskutiert. Weiterhin wurde der Gehalt von Mn, Co, Ni, Cu, Zn, Cd, Hg, Pb im Fleisch von Miesmuscheln (Mytilus galloprovincialis Lamarck) untersucht. Die Mollusken stammen aus der Umgebung des Hafens von Triest, in der Nähe eines wichtigen Abwasserkanals. Die Korrelationsmatrix geht aus der 8 × 43-Datenmatrix nach einer logarithmischen Transformation der Konzentrationsvariablen hervor, dies basiert auf der Analyse der Hauptkomponenten. 8 Variable wurden auf 4 Hauptkomponenten, die 80% der totalen Varianz erklären, reduziert. Die orthogonal-rotierte Faktorenmatrix zeigt, daß Co, Ni, Cd und Pb an die erste Hauptkomponente, Cu und Zn an die zweite, Hg an die dritte und Mn an die vierte Hauptkomponente gebunden sind. Die Ergebnisse dieser Forschung werden mit denen vorhergehender mehrdimensionaler Analysen aus zwei Gegenden in der Bucht von Muggia verglichen. Die Herkunft der Spurenelemente im Muschelfleisch aus dem Golf von Triest werden diskutiert.

     

Surface lead pollution of roadside crops in relation to the distance from an emitting line source

The asymptotic decrease in lead concentration c in roadside crops according to the distance d from the edge of an isolated straight road can be described empirically by means of the three-parameter exponential function c=A+B exp (-Cd). Least-squares non-linear regression allows the estimation of the parameters A, B and C. The validity of the function was tested by fitting it to a few sets of observations on unwashed wine grape and leaf samples. An analysis of residuals was performed, considering in particular the extent of the normality of their distribution. The function was also applied to a few sets of data of low sample size to examine the effect of washing on the parameters of the function.     

Aligned Nanofiber Topographies Enhance the Differentiation of Adult Renal Stem Cells into Glomerular Podocytes

The use of adult stem/progenitor cells is a challenge in current research about kidney functional regeneration. In this framework, material‐induced stem cell differentiation can become a new paradigm to promote advances in therapies. Here, the effects of both isotropic and anisotropic fibrous topographies on the podocyte differentiation of adult renal stem cells (RSCs) from human donors are investigated. The proliferation rate of RSCs is analyzed, immunofluorescence and genetic analyses of specific podocyte markers (Wilms’ tumor 1 gene, nephrin, and podocin) are performed to assess the differentiation level on the scaffolds. The studied markers are over‐expressed in RSCs cultured on aligned fibers compared to cells cultured on either protein‐functionalized films or randomly oriented fibers. In addition, RSCs cultured on aligned fibers are found to differentiate toward podocyte precursors even in basal medium conditions, thus highlighting scaffold‐induced commitment without exogenous chemicals or cellular reprogramming. Aligned polymer fiber scaffolds which provide instructive cues for RSC differentiation might lead to new biomimetic systems for renal stem cell engineering.