A combined protocol with piroxicam,chemotherapy, and whole pelvic irradiation with simultaneous boost volumetric modulated arc radiotherapy for muscle-invasive canine urinary transitional cell carcinoma: First clinical experience

The aims of this pilot study were to evaluate the feasibility and efficacy of high-dose hypofractionated volumetric modulated arc radiotherapy (VMAT) applied to whole pelvic region radiotherapy (WPRT) with multilevel simultaneous integrated boost (MLSIB) combined with piroxicam and chemotherapy for the treatment of canine transitional cell carcinoma (TCC) of the lower urinary tract with muscle invasion TCC. Twelve dogs were enrolled, according to stage, in two groups: group 1, TCC confined to the urinary tract; group 2, TCC with metastasis. The planning target volume dose was tailored from 36 to 42 Gy in 6 fractions. All dogs were prescribed piroxicam and radiosensitizing carboplatin, and six received chemotherapy after radiotherapy. Serial follow-ups with computed tomography and magnetic resonance imaging were performed. Disease control and toxicity effects were evaluated according to the Response Evaluation Criteria in Solid Tumors and Veterinary Radiation Therapy Oncology Group criteria. The treatment was well tolerated, and no high-grade side effects were reported. The median overall survival times for groups 1 and 2 were 1,230 and 150 days, respectively. A considerable percentage of patients in group1 (50%) were still alive at the time of writing this paper, and a longer follow-up could enable a more accurate survival analysis. This preliminary analysis shows that VMAT applied to the WPRT with MLSIB is an effective and safe option for dogs with lower urinary TCC, although the presence of metastases worsens the prognosis.     

Prey depletion caused by overfishing and the decline of marine megafauna in eastern Ionian Sea coastal waters (central Mediterranean)

Surveys primarily aimed at determining dolphin encounter rates were conducted from small inflatable craft in eastern Ionian Sea coastal waters between 1997 and 2004. During 633 surveys totalling 21,276 km of effort, observations of cetaceans and other marine species spotted in a study area of 480 km² were systematically recorded. Common dolphin encounter rates declined 25-fold across the study period, steadily decreasing from 2.18 encounters/100 km in 1997 to 0.09 encounters/100 km in 2004. Encounter rates of tuna also declined significantly. Swordfish encounter rates dropped from 1.03 encounters/100 km in 1997 to 0-0.12 in 1998-2004. Encounter rates of bottlenose dolphins did not show significant trends. The decline of high-order marine predators feeding on epipelagic prey was consistent with the hypothesis of prey depletion, likely resulting from intensive exploitation of local fish stocks, particularly anchovies and sardines. The catholic feeding habits and opportunistic behaviour of bottlenose dolphins may allow them to withstand the effects of overfishing at their present low density.     

Monitoring organic matter dynamics in soil profiles by 'Rock-Eval pyrolysis': bulk characterization and quantification of degradation

Rock‐Eval pyrolysis was designed for petroleum exploration to determine the type and quality of organic matter in rock samples. Nevertheless, this technique can be used for bulk characterization of the immature organic matter in soil samples and recent sediments. We studied 76 samples from seven soil classes and showed that their pyrograms can be described by a combination of four elementary Gaussian components: F1, F2, F3 and F4. These four components are related to major classes of organic constituents differing in origin and their resistance to pyrolysis: labile biological constituents (F1), resistant biological constituents (F2), immature non‐biotic constituents (F3) and a mature refractory fraction (F4). We discriminated the relative contributions of these components and used them to derive two indices: (i) to quantify the relative contributions of labile and resistant biological constituents and (ii) to quantify the degradation stage of the soil organic matter. The practical applications are illustrated via the influence of vegetal cover on soil organic matter dynamics and peat development in a Holocene sedimentary sequence, but we suggest that the approach is of much wider application.     

Simple and rapid method for the differentiation of Lens culinaris Medik. from false lentil species

The practice of using both common vetch (Vicia sativa L.) and single-flower vetch (Vicia articulata Hornem.) seeds as food or feed is encouraged by the very high resemblance of their seeds with those of small-seeded lentil cultivars. Among the Vicieae, antinutritional and toxic factors are particularly important, because many species, containing high levels of these compounds, are not safe. A simple and fast capillary electrophoresis (CE) method was proposed for the differentiation of lentil cultivars from false lentil species (i.e., single-flour vetch and common vetch). Proteins were extracted from defatted milled seeds with an alcoholic/saline solution. Extracts were separated in an uncoated fused-silica capillary with iminodiacetic acid (IDA) isolectric buffer containing 0.05% hydroxypropylmethylcellulose (HPMC) and 20% acetonitrile. The presented method is useful also for the detection of contamination of whole or split seeds of lentil by vetch species. With respect to alternative techniques, such as DNA-based markers or thin-layer chromatography (TLC), CE has the advantages of being less expensive, faster, and fully automated.     

Nondestructive evaluation of anthocyanins in olive (Olea europaea) fruits by in situ chlorophyll fluorescence spectroscopy

Anthocyanins (Anths) in olive (Olea europaea L.) fruits at different degrees of pigmentation were assessed nondestructively by measuring chlorophyll fluorescence (ChlF). The method is based on the comparison of the ChlF excitation spectra from olives with different pigmentation from green to green-red, reddish-purple, and purple. The logarithm of the ratio between the fluorescence excitation spectra (logFER) from two different colored zones gave the difference in the absorption spectrum between them. The absorbance spectrum derived from the logFER between a red olive and the same olive devoid of the skin showed the typical Anth green band (at 550 nm). It matched that recorded by microspectrophotometry on a single pulp cell and the in vitro absorbance spectrum of the olive skin extract. As expected, the in vivo Anths absorption maximum increased in intensity going from less to more mature olives and was higher in the sun-exposed olive side with respect to the sun-shaded side. Absolute quantitative nondestructive determination of Anths for each olive sample was obtained by the logFER calculated for two excitation wavelengths, 550 and 625 nm, of ChlF at 740 nm. Going from green to purple skin colors, the Log[ChlF(625)/ChlF(550)] was fairly well-correlated to the extract Anths concentration. Finally, the relationship between the Anths and the other main phenolics present in the olives analyzed by high-performance liquid chromatography was evaluated. The main result was a net increase of verbascoside with increasing Anths content. On the basis of our results, the development of a new rapid and noninvasive method for the monitoring of olive development and ripening can be envisaged.     

Stable isotope characterization of the ortho-oxygenated phenylpropanoids: coumarin and melilotol

The natural abundance 2H NMR spectra of extractive coumarin 10 and of its dihydroderivative melilotol 11 produced by baker's yeast reduction has been compared with synthetic materials. Diagnostic for the differentiation of 10 are the (D/H)beta values, which are in the 128.1-133.6 ppm interval for the natural compounds but 258.5 and 189.8 ppm for the synthetic materials. Such a dramatic difference is also found for methyl cinnamate 12, which shows (D/H)beta values of 127.2 and 515.8 ppm, respectively. In extractive 10, the ratio (D/H)4para/(D/H)6ortho = 1.24 is similar to that observed in structurally related salicin and methyl salicylate. Coumarin 10 is transformed in salicyl alcohol 9, providing diacetate 14, showing in the natural series the trend (D/H)3meta > (D/H)4para > (D/H)5meta approximately (D/H)6ortho. A similar trend is shown also by the synthetic 10. A clear distinction between extractive and synthetic 10 is obtained through delta18O determinations on 10 and on chroman 13. The bulk delta18O values in the extractive series of 10 are 20.3, 23.6, and 22.6 per thousand, while those of the aromatic oxygen are 2.3, 0.5, and -0.5 per thousand. In the synthetic sample, the values are 12.6 and 5.6 per thousand, respectively. As a final product, the reduction of 10 leads to the dihydroderivative 11. Both the baker's yeast reduction and the catalytic hydrogenation lead to a marked decrease of the deuterium content of 11, which is stronger for the beta-position than for the alpha-position.     

Fast determination of catechins and xanthines in tea beverages by micellar electrokinetic chromatography

Antioxidant properties and stimulating effects of green tea are related to its content of cathechins and xanthines; tea quality evaluation is based on organoleptic tests and on the presence of those components. In this work, by a MEKC method, eight cathechins and three xanthines were quantified in some tea-based beverages. The best separation was realized using a phosphate-borate running buffer, with sodium dodecyl sulfate as micellar agent. A 40 cm capillary, a temperature of 29 degrees C, a voltage of 30 kV, and UV detection at 200 nm were used. The method showed a very good sensitivity (limit of detection ranging from 0.0011 to 0.0051 microg/mL) and was applied to real tea samples to characterize their antioxidant content. Statistical studies were performed and showed a satisfactory reliability of the data.     

Stable isotope labeling pattern of resveratrol and related natural stilbenes

The stable isotope characterization of resveratrol 1 from Polygonum cuspidatum and of related natural stilbenes 11 and 12 obtained by hydrolysis of the corresponding glucosides 2 and 3 from Rheum is reported. The C(6)-C(2)-C(6) framework of suitably protected derivatives of 1, 2, and 3 has been degraded with ozone to the C(6)-C(1) aldehydes 4, 5, 9, and 10, retaining all hydrogen atoms of the precursors. The natural and synthetic derivatives are characterized and distinguished by natural abundance deuterium nuclear magnetic resonance studies. In the case of anisaldehyde 4 the two series show, as expected, the characteristic difference of the aromatic labeling. The formyl deuterium contents of 4 and 5 from resveratrol are remarkably different, seemingly reflecting the different enrichments existing between positions 3 and 2, respectively, of the phenylpropanoid precursor. The positional delta(18)O values of the extractive materials 1-3 were also determined. In this instance a selective deoxygenation procedure was adopted, leading from 1 to the products 6, 7, and 8. The delta(18)O values of the latter compounds reveal, respectively, those at position 4' and positions 3 and 5 of 1. Similarly, the phenolic products 11 and 12 were converted into 13 and 14. From the delta(18)O values of the single components it is possible to design a detailed map of the oxygen fractionations which characterizes the stilbenes 1-3. In particular, the oxygen present at position 4' of the phenylpropanoid moiety of 1-3 shows delta(18)O values of +11.5, +1.8, and +6.7 per thousand, respectively. Moreover, the phenolic oxygen atom at position 3' of rhapontin 3 shows a value of +11.7 per thousand. The data are compared with those previously obtained on structurally related compounds. These results show the utility of simple chemical degradations in the stable isotope characterization of structurally complex food components.     

Compositional shifts of bacterial groups in a solarized and amended soil as determined by denaturing gradient gel electrophoresis

Soil solarization is a widespread, nonchemical agricultural practice for disinfesting soils, which is often used in combination with organic amendment, and whose action represents an important factor impacting on soil bacterial communities structure and population dynamics. The present study was conducted to investigate whether and to which extent a 72-day plot-scale soil solarization treatment, either combined or not with organic amendment, could stimulate compositional changes in the genetic structure of indigenous soil bacterial communities. Soil solarization with transparent polyethylene film, in combination or not with farmyard manure addition, was carried out during a summer period on a clay loam agricultural soil located in Southern Italy. Soils from a four-treatment (NS, nonsolarized control soil; S, solarized soil; MA, manure-amended nonsolarized soil; MS, manure-amended and solarized soil) plot block were sampled after 0, 8, 16, 36 and 72 days. Compositional shifts in the genetic structure of indigenous soil bacterial communities were monitored by denaturing gradient gel electrophoresis (DGGE) fingerprinting of 16S rRNA gene fragments amplified from soil-extracted community DNA using primers specific for Bacteria, Actinomycetales, α- and β-Proteobacteria. Changes in soil temperature, pH, and electrical conductivity (EC_1:1) were also monitored from 0 to 72 days. Beneath the polyethylene film the average soil temperature at 8-cm depth reached 55 °C compared to 35 °C in nonsolarized soil. In general, without amendment both soil pH and EC_1:1 were not significantly affected by solarization, whereas in manured plots either variables were greatly increased (from 7.0 to 8.0 pH and from 271 to 3021 μS cm⁻¹ EC_1:1), and both showed long-lasting effects due to soil solar heating. The eubacterial DGGE profiles revealed that soil solarization was the main factor inducing strong time-dependent population shifts in the community structure either in unamended or amended soils. Conversely, the addition of organic amendment resulted in an altered bacterial community, which remained rather stable over time. A similar behaviour was also observed in the DGGE patterns of β-proteobacterial and actinomycete populations, and also, albeit to a lesser extent, in the DGGE profiles of α-Proteobacteria. An increased bacterial richness was evidenced by DGGE fingerprints in 16- and 36-day samplings, followed by a decrease appearing in 72-day samplings. This could be explained, other than by a direct thermal effect on soil microflora, by solarization-induced changes in the physico-chemical properties of soil microbial habitats or by other ecological factors (e.g. decreased competitiveness of dominating bacterial species, reduced grazing pressure of microfaunal predators, increased nutrient availability).     

Anthracene influence on (15NH4)2SO4 fertilization of wheat seedlings

The influence of the addition of anthracene (1 μg anthracene g^?1 soil) in N transformations following (¹⁵NH₄)₂SO₄ fertilization (200 mg N g^?1 soil) was investigated in wheat pots by quadrapole mass-spectrometry. The dry matter yield at harvesting (after 16 days) was not statistically affected (P=0.05) by anthracene addition. The total amount of N from the fertilizer taken up by wheat seedings in 16 days was 29 and 26.8% of the added N in the absence and in the presence of anthracene, respectively, but the difference was not significantly different at level P=0.05. In order to investigate more deeply the effect of anthracene on the N cycle in the soil-plant system, the first-order rate constants of N mineralization, N immobilization, nitrification and N plant uptake have been determined according to a ¹⁵N + ¹⁴N soil-plant model. The comparison of the constants showed that organic N mineralization, nitrification and plant uptake proceeded at the same rate, while a small different rate (P=0.05) was shown by N immobilization. In fact, the N immobilization constant increased from 0.14±0.012 to 0.21±0.014 day^?1 as a consequence of anthracene addition.