Response of rainbow trout gill (Na++K+)-ATPase and chloride cells to T3 and NaCl administration

With the aim of comparing the effects of oral T₃ and NaCl administration on trout hypoosmoregulatory mechanisms, three groups of rainbow trout (Oncorhynchus mykiss Walbaum) held in freshwater (FW) were fed a basal diet (C), the same diet containing 8.83 ppm of 3,5,3-triiodo-L-thyronine (T₃) (T) or 10% (w/w) NaCl (N) respectively for 30 d. They were then transferred to brackish water (BW) for 22 d and fed on diet C. Gill (Na⁺+K⁺)-ATPase activity and its dependence on ATP, Na⁺ and pH, number of gill chloride cells (CC), serum T₃ level as well as fish growth, condition factor (K) and mortality were evaluated. During the FW phase, as compared to C trout, T trout showed a two fold higher serum T₃ level, had unchanged gill (Na⁺+K⁺)-ATPase activity and increased CC number, whereas N trout showed higher gill (Na⁺+K⁺)-ATPase activity and CC number. At the end of the experiment the enzyme activity was in the order T>N>C groups and all groups showed similar CC number. Both treatments changed the enzyme activation kinetics by ATP and Na⁺. A transient increase in K value occurred in N group during the period of salt administration. In BW, T and N groups had higher and lower survival than C group respectively. Other parameters were unaffected by the treatments. This trial suggests that T₃ administration promotes the development of hypoosmoregulatory mechanisms of trout but it leaves the (Na⁺+K⁺)-ATPase activity unaltered till the transfer to a hyperosmotic environment.     

Toward metrological traceability for DNA fragment ratios in GM quantification. 1. Effect of DNA extraction methods on the quantitative determination of Bt176 corn by real-time PCR

An international CCQM-P60 pilot study involving eight national metrological institutes was organized to investigate if the quantification of genetically modified (GM) corn powder by real-time PCR was affected by the DNA extraction method applied. Four commonly used extraction methods were compared for the extraction of DNA from a GM Bt176 corn powder. The CTAB-based method yielded the highest DNA template quantity and quality. A difference in the 260 nm/230 nm absorbance ratio was observed among the different extraction methods. Real-time amplification of sequences specific for endogenous genes zein and hmg as well as transgenic sequences within the cryIA(b) gene and a fragment covering the junction between the transformed DNA and the plant genome were used to determine the GM percentage. The detection of the transgenic gene was affected by the quantity and quality of template used for the PCR reaction. The Bt176 percentages measured on diluted or purified templates were statistically different depending on the extraction method applied.     

Comparison of different tissue sampling for PCR-based diagnosis and follow-up of canine visceral leishmaniosis

In this study, different types of tissue sampling for PCR-based diagnosis and follow-up of canine visceral leishmaniosis were compared. Skin, whole blood and lymph node samples were collected from 95 naturally infected dogs living in South Italy, where the disease is endemic. Twenty-nine of these 95 dogs, treated with meglumine administered concurrently with allopurinol for 30 days, and then with allopurinol alone, were monitored during a period of 2 years. The DNA extracted from the clinical specimens was amplified by PCR using as target DNA a 116-bp fragment in the constant region of the kinetoplast DNA minicircle. PCR analysis was more sensitive than indirect immunofluorescence antibody test in detecting Leishmania infection in symptomatic dogs: 99% of lymph node samples resulted positive, whereas 94% of blood samples and 95% of skin samples gave a positive result. PCR analysis of samples from dogs followed up 2 years showed that: (1) all subjects resulted positive in at least one of the three types of samples; (2) all time the dogs had a relapse, PCR resulted positive in all three types of samples; (3) when dogs were apparently healthy, PCR analysis was positive on skin and lymph node samples, but not always on blood samples. Since lymph node sampling is invasive and sometimes difficult in healthy asymptomatic dogs, our results suggest that, independently from the presence or not of cutaneous lesions, skin biopsy represents a good substratum for PCR-based diagnosis and follow-up of canine visceral leishmaniosis.     

Morpho‐physiological variation of Poa alpina L. genetic resources from the Rhaetian Alps,Italy, grown in two altitude‐contrasting sites

Alpine bluegrass (Poa alpina L.) is common in mountain areas, where it represents a valuable forage resource and is widely used for ecological restoration of disturbed landscapes. This study assessed the variation of germplasm from the Rhaetian Alps, Italy. Both seed production and morphology were examined for potential use in restoration projects and for conservation and selection purposes of local genetic resources. The effects of the ex situ growing site were also assessed in two altitude‐contrasting sites. Individual populations harboured great phenotypic diversity for all traits, including seed yield, while variation among valleys of origin was less manifest. Noticeable site‐specific population yield responses were observed, suggesting that the environment adopted for seed production may steer the choice of materials to be multiplied. The longer vegetative period in the lowland site, combined with environment‐tailored agronomic practices, contributed to better plant growth, seed yield and survival compared to the mountain site, suggesting that seed production of this Alpine species could be feasible in a lowland environment when adequate inputs were provided. One valley (Valchiavenna) displayed high average levels of seed yield and plant vigour across growing sites, indicating its possible value as a source of germplasm suitable for resowing at high altitude.     

Boron content in freshwaters of Northern Italy

The B concentration was determined in bulk deposition and in surface freshwaters (lakes and rivers) of the river Po watershed in Northern Italy. The curcumin photometric method was used to determine B content for all analyses. The B concentrations were under detection limits (0.06 mg B L^?1) in bulk deposition and below 0.09 mg B L^?1 in lake waters. Approximately 65 % of river samples measured had B concentrations close to natural background levels for natural waters (0.1 mg B L^?1). There was a strong correlation (r < 0.85) between B concentration and those of both total dissolved P and anionic detergents. The elevated B concentrations may be related to anthropogenic sources.     

Using Monte Carlo simulations to estimate relative fire ignition danger in a low-to-medium fire-prone region

A comprehensive assessment of fire ignition danger is nowadays a basic step towards the prioritization of fire management measures. In this study we propose performing a fire selectivity analysis using Monte Carlo simulations to statistically estimate the relative fire ignition danger in a low-to-intermediate fire-prone region such as Canton Ticino, Switzerland. We define fire ignition danger as the likelihood that at a given place a fire will be ignited. For each 25 m × 25 m pixel of the study area, landscape characteristics that may be related to the probability of fire ignition such as vegetation type, elevation, aspect, slope, urban-forest interface were first split into 9-12 categories. The selectivity of each category with respect to fire ignition was then statistically tested by means of Monte Carlo simulations. Finally, we proposed two different approaches for calculating the ignition danger index: cumulating the scores of the Monte Carlo simulations to a final index or producing synthetic scores by performing a principal component analysis of the Monte Carlo results. The validation of the resulting fire danger indices highlights the suitability of both proposed approaches. The PCA-option allows a slightly better discrimination between ignition and non-ignition points and may be of more general application.     

Chemical composition and antimicrobial activity of the essential oil from Ferula glauca L. (F. communis L. subsp. glauca) growing in Marche (central Italy)

The essential oil obtained from different parts of Ferula glauca L. (formerly considered as a subspecies of F. communis) growing in Marche (central Italy), was analyzed for the first time by GC-FID and GC-MS. The major volatiles were (E)-caryophyllene and caryophyllene oxide in leaves, alpha-pinene, myrcene and germacrene D in flowers, alpha- and beta-pinene in fruits, (E)-beta-farnesene, myristicin and elemicin in roots, respectively. The differences in composition detected with respect to F. communis, made the volatile fraction a reliable marker to distinguish between them, and confirm the botanical data at the base of their discrimination. Furthermore, the oil was assayed for its antimicrobial activity by the broth microdilution method. B. subtilis was found to be the most sensitive microorganism, with the lowest MIC values.     

Benzodioxole derivatives as negative effectors of plant proteases

Previous work demonstrated that a commercial formulation of piperonyl butoxide (PBO) did inhibit the activity of some plant proteolytic enzymes. In this paper, the effect of pure PBO and nine pure PBO homologues (PBOH) appropriately synthesized toward bromelain and papain was studied in hydrocarbon solution using the bis(2-ethylhexyl)sodium sulfosuccinate (AOT) reverse micellar system. This study establishes that the majority of these compounds show, in vitro, interesting protease inhibition activities. The benzodioxole and dihydrobenzofuran structures, in particular, 5-[2-(2-butoxyethoxy)ethoxymethyl]-benzo[1,3]dioxole (EN 1-40) and 6-[2-(2-butoxyethoxy)ethoxymethyl]-5-propyl-2,3-dihydrobenzofuran (EN 16-5), respectively, appear to be responsible for protease inhibition. Measures of octanol/water partition coefficients on PBO and PBOH have demonstrated that water solubility plays a fundamental role in the expression of protease inhibition activity.     

Blanching Improves Anthocyanin Absorption from Highbush Blueberry ( Vaccinium corymbosum L.) Purée in Healthy Human Volunteers: A Pilot Study

Blueberries ( Vaccinium corymbosum L.) are rich sources of phenolics and anthocyanins (ACNs). We investigated the absorption of ACNs after consumption of one portion (300 g) of minimally processed blueberry purée (P) obtained from blanched (BL) or unblanched (NB) berries. A repeated-measures, crossover design study was conducted on healthy human volunteers. Blood was drawn between baseline and 24 h after BL-P or NB-P consumption, while urine were collected from the day before the experiment up to 48 h. Total plasma ACN content was not significantly different, while phenolics content was higher in BL-P with respect to NB-P. The maximum ACN absorption in plasma was observed after 1.5 h from the intake of the purées and was significantly higher (p ≤ 0.05) after the intake of BL-P. Both products increased the excretion of hippuric acid in urine. In conclusion, blanching had no significant effect on total ACN content and enhanced their absorption from minimally processed purées.