In vitro study of the replication capacity of the RGNNV and the SJNNV betanodavirus genotypes and their natural reassortants in response to temperature

Betanodaviruses are the causative agents of viral nervous necrosis and affect a broad range of fish species worldwide. Their bi-segmented genome is composed of the RNA1 and the RNA2 molecules encoding the viral polymerase and the coat protein, respectively. In southern Europe the presence of the RGNNV and the SJNNV genotypes, and the RGNNV/SJNNV and RGNNV/SJNNV reassortants has been documented. Several studies have reported a correlation between water temperature and disease onset. To explore the replication efficiency of betanodaviruses with different genomes in relation to temperature and to understand the role of genetic reassortment on viral phenotype, RGNNV, SJNNV, RGNNV/SJNNV and RGNNV/SJNNV field isolates were fully sequenced, and growth curves generated in vitro at four different temperatures (15, 20, 25, 30 °C) were developed for each isolate. The data obtained, corroborated by statistical analysis, demonstrated that viral titres of diverse betanodavirus genotypes varied significantly in relation to the incubation temperature of the culture. In particular, at 30 °C betanodaviruses under investigation presented different phenotypes, and viruses containing the RNA1 of the RGNNV genotype showed the best replication efficiency. Laboratory results demonstrated that viruses clustering within the same genotype based on the polymerase gene, possess similar growth kinetics in response to temperature, thus highlighting the key role of RNA1 in controlling viral replication at different environmental conditions. The results generated might have practical implications for the inference of viral phenotype according to genetic features and may contribute to a better understanding of betanodavirus ecology.     

IMAGING DIAGNOSIS—TRANSESOPHAGEAL ULTRASOUND‐GUIDED REMOVAL OF A MIGRATING GRASS AWN FOREIGN BODY IN A DOG

A 3‐year‐old English Setter dog was presented for an acute onset of coughing. Tracheobronchoscopic examination allowed localization and removal of one grass awn foreign body. A second migrated grass awn was suspected to be present in the left caudal lung lobe. Transesophageal ultrasound revealed an area of pulmonary consolidation in the dorsomedial portion of left caudal lobe and a linear hyperechoic structure consistent with a grass awn foreign body within the area of consolidation. Transesophageal ultrasonography was also used to provide anatomical landmarks that facilitated successful thoracoscopic removal of the foreign body.     

Collapse of a degenerate Fermi gas

A degenerate gas of identical fermions is brought to collapse by the interaction with a Bose-Einstein condensate. We used an atomic mixture of fermionic potassium-40 and bosonic rubidium-87, in which the strong interspecies attraction leads to an instability above a critical number of particles. The observed phenomenon suggests a direction for manipulating fermion-fermion interactions on the route to superfluidity.     

CB1 cannabinoid receptors and on-demand defense against excitotoxicity

Abnormally high spiking activity can damage neurons. Signaling systems to protect neurons from the consequences of abnormal discharge activity have been postulated. We generated conditional mutant mice that lack expression of the cannabinoid receptor type 1 in principal forebrain neurons but not in adjacent inhibitory interneurons. In mutant mice,the excitotoxin kainic acid (KA) induced excessive seizures in vivo. The threshold to KA-induced neuronal excitation in vitro was severely reduced in hippocampal pyramidal neurons of mutants. KA administration rapidly raised hippocampal levels of anandamide and induced protective mechanisms in wild-type principal hippocampal neurons. These protective mechanisms could not be triggered in mutant mice. The endogenous cannabinoid system thus provides on-demand protection against acute excitotoxicity in central nervous system neurons.     

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.