NECROTIZING ENCEPHALITIS IN A YORKSHIRE TERRIER: CLINICAL, IMAGING, AND PATHOLOGIC FINDINGS

A 5-year-old, male Yorkshire Terrier had chronic and progressive neurologic signs compatible with lesions in the right brain stem and right forebrain. In magnetic resonance images of the brain there were multifocal lesions at different stages of evolution, consistent with an inflammatory disease. The lesions were located in the white matter of the cerebrum, in the diencephalon and mesencephalon. These lesions were hypointense in T1-weighted images and hyperintense in T2-weighted images with no mass effect and minimal enhancement with gadolinium. Necrotizing encephalitis was confirmed post mortem.     

Mixotrophy in a Local Strain of Nannochloropsis granulata for Renewable High-Value Biomass Production on the West Coast of Sweden

A local strain of Nannochloropsis granulata (Ng) has been reported as the most productive microalgal strain in terms of both biomass yield and lipid content when cultivated in photobioreactors that simulate the light and temperature conditions during the summer on the west coast of Sweden. To further increase the biomass and the biotechnological potential of this strain in these conditions, mixotrophic growth (i.e., the simultaneous use of photosynthesis and respiration) with glycerol as an external carbon source was investigated in this study and compared with phototrophic growth that made use of air enriched with 1–2% CO₂. The addition of either glycerol or CO₂-enriched air stimulated the growth of Ng and theproduction of high-value long-chain polyunsaturated fatty acids (EPA) as well as the carotenoid canthaxanthin. Bioassays in human prostate cell lines indicated the highest antitumoral activity for Ng extracts and fractions from mixotrophic conditions. Metabolomics detected betaine lipids specifically in the bioactive fractions, suggesting their involvement in the observed antitumoral effect. Genes related to autophagy were found to be upregulated by the most bioactive fraction, suggesting a possible therapeutic target against prostate cancer progression. Taken together, our results suggest that the local Ng strain can be cultivated mixotrophically in summer conditions on the west coast of Sweden for the production of high-value biomass containing antiproliferative compounds, carotenoids, and EPA.     

Investigating landscape phase transitions in Mediterranean rangelands by recurrence analysis

Context

Socio-ecological landscapes typically characterized by non-linear dynamics in space and time are difficult to be analyzed using standard quantitative methods, due to multiple processes interacting on different spatial and temporal scales. This poses a challenge to the identification of appropriate approaches for analyzing time series that can evaluate system properties of landscape dynamics in the face of disturbances, such as uncontrolled fires.

Objective

The purpose is the application of non-linear methods such as recurrence quantification analysis (RQA) to landscape ecology. The examples concern the time series of burnt and unburnt Mediterranean rangelands, to highlight potential and limits of RQA.

Methods

We used RQA together with joint recurrence analysis (JRA) to compare the evolutionary behavior of different land uses.

Results

Time series of forests and grasslands in rangelands present both periodic and chaotic components with a rather similar behavior after the fire and clear transitions from less to more regular/predictable dynamics/succession. Results highlight the impacts of fire, the recovery capacity of land covers to pre-burnt levels, and the decay of synchronization towards the previous regime associated with vegetation secondary succession consistent with early successional species.

Conclusions

RQA and JRA with their set of indices (recurrence rate: RR, laminarity: LAM, determinism: DET, and divergence: DIV) can represent new sensitive measures that may monitor the adaptive capacity and the resilience of landscapes. However, future applications are needed to standardize the analysis by strengthening the accuracy of this approach in describing the ongoing transformations of natural and man-managed landscapes.

     

Using energy balance data for assessing evapotranspiration and crop coefficients in a Mediterranean vineyard

Improving water use efficiency is a key element of water management in irrigated viticulture, especially in arid or semi-arid areas. In this study, the micrometeorological technique “Eddy Covariance” was used to directly quantify the crop evapotranspiration (ET) and to analyze the complex relationships between evapotranspiration, energy fluxes, and meteorological conditions. Both observed Direct measurements (DIR) of latent heat flux (LE) and observed from the residual of the energy balance (REB) equation were used for crop evapotranspiration calculations. Observed crop coefficients (K _cms) were then determined using the standardized reference evapotranspiration (ET_o) equation for short canopies. In addition, linear approximations from observations were used to model the seasonal trend lines for crop coefficients and K _cs values were parameterized by first identifying the beginning and end of each growth stage. The modeled K _cs values were used to predict daily ET from ET_o measurements and compared with values from literature. The daily observed DIR ET values (ET_do) were lower than REB ET (ET_ro) during periods with precipitation, but they were similar during dry periods, which implies that energy balance closure is better when the surface is drier. Comparisons between modeled ET and crop ET estimated using K _c values from best agreement was observed between the modeled REB and FAO 56 and the local K _c values provided by the Regional Agency ARPAS showed good agreement with observed ET (from DIR and REB data) than the FAO 56 ones. The study confirmed that the availability of locally driven K _c could be relevant to quantify the crop water requirement and represents the starting point for a sustainable management of water resources.     

Phenolic antioxidants and the protection of low density lipoprotein from peroxynitrite-mediated oxidations at physiologic CO2

Dietary phenolic antioxidants have been shown to prevent LDL modifications mediated by several physiologic oxidants including peroxynitrite. However, more recent data demonstrated that CO(2) affected the fate of peroxynitrite in biological fluids and significantly reduced peroxynitrite scavenging by polyphenols, raising doubts concerning their antioxidant activity. We found that the oxidation of LDL lipids mediated by peroxynitrite decreased in the presence of bicarbonate, while Trp oxidation and 3-nitroTyr formation increased, suggesting a redirection of peroxynitrite reactivity toward the protein moiety. We therefore evaluated the protective activity of some phenolic antioxidants (quercetin, oleuropein, resveratrol, (+)-catechin, (-)-epicatechin, tyrosol, alpha- and gamma-tocopherol, ascorbate) on peroxynitrite-mediated oxidation of LDL aromatic residues. Some of these phenols protected LDL Trp from oxidation better than ascorbate or alpha-tocopherol, although protection at 100 microM did not exceed 30-40%. However, the same phenolic antioxidants were more active in inhibiting 3-nitroTyr formation and those with a catechin structure provided significant protection (IC(50%) 40-50 microM). Red wine, a polyphenol-rich beverage, showed a protective effect comparable to that of the most active phenolic antioxidants. Direct EPR studies showed that bicarbonate significantly increased the peroxynitrite-dependent formation of O-semiquinone radicals in red wine, supporting the hypothesis that polyphenols are efficient scavengers of radicals formed by peroxynitrite/CO(2). Ascorbate was a poor inhibitor of peroxynitrite/CO(2)-induced LDL tyrosine nitration, but the simultaneous addition to the most active polyphenols halved their IC(50%). In conclusion, although cooperation with other antioxidants can further decrease the IC(50%) of polyphenolics, as demonstrated for ascorbate, their antioxidant activity appears to occur at concentrations at least 1 order of magnitude higher than their bioavailability.     

Assay of riboflavin in sample wines by capillary zone electrophoresis and laser-induced fluorescence detection

To routinely assay the concentration of riboflavin (RF) in wines, a rapid and sensitive method was developed and evaluated. The method is based on a simple sample preparation, capillary zone electrophoretic separation and laser-induced fluorescence detection (CZE-LIF). Sample preparation required only dilution and filtration. Under optimized conditions, the limit of detection of riboflavin was 0.5 micro g/L, using a hydrodynamic sample introduction of 10 s at 54 mbar. The method was fully validated: the recovery of RF in wines was >95%. The concentrations of RF within the three sample types of Italian wines investigated here ranged from 69 to 151 micro g/L with a mean value (+/-SD) of 112 +/- 25 micro g/L, from 74 to 193 micro g/L with a mean value of 115 +/- 45 micro g/L, and from 156 to 292 micro g/L with a mean value of 226 +/- 40 micro g/L, for white, rosé, and red wines, respectively. Such an accurate and highly sensitive CZE-LIF method represents a powerful improvement over previous methods in terms of sensitivity, simplicity, and efficiency. It is well suited to satisfy the demands for accurate and sensitive detection with minimal sample preparation and cleanup.     

Diversity of heterotrophic aerobic cultivable microbial communities of soils treated with fumigants and dynamics of metabolic, microbial, and mineralization quotients

A combination of molecular and classical techniques was used to study the composition, structure, diversity, and dynamics of an aerobic heterotrophic cultivable bacterial community isolated from five different soil samples treated with the fumigant agent 1,3-dichloropropene (1,3-D) and further subjected to nitrogen–phosphorous–potassium (NPK) fertigation (F), amendment (C ₂ and C ₄), and NPK fertigation plus amendment (F + C) in two different periods (May and July). The restriction and sequence analysis of 16S rDNA from 189 isolates revealed a very high percentage (94%) of Gram-positive bacterial isolates, most of which (83%) belonging to the genus Bacillus. The degree of intraspecific genetic diversity was high, as shown by random amplified polymorphic DNA analysis. These data seem to be related with the increase in microbial biomass C (C _mic) content and the decrease in the total organic C (C _org) and metabolic quotient (qCO₂) values, especially in amended soils (C ₂, C ₄) where soil microflora mineralized the organic matter of the added fertilizers. In a short term, it is suggested that the presence of very high percentage of Gram-positive bacteria might be related to the ability of these bacteria to form spores so as to be resistant to fumigants rather than being the result of a selective pressure in the predominance of microbial species with a set of genes involved in biodegradation of 1,3-D. Stefano Mocali and Donatella Paffetti contributed equally to this work.     

Determination of free proline and monosaccharides in wine samples by high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD)

A sensitive and selective analytical method for the simultaneous separation and quantitative determination of proline and free monosaccharides in wine samples by high-performance anion-exchange chromatography coupled with pulsed amperometric detection is described. Under optimized experimental conditions, a complete separation was obtained in less than 30 min, using an isocratic elution with 10 mM NaOH and 1 mM Ba(OAc)(2). No postcolumn addition of strong bases to the eluent for enhancing detection sensitivity was needed. Upon 25-fold sample dilution and purification to avoid interference of tannins, pigments, and phenolic compounds, the fingerprinting of common monosaccharides (i.e., arabinose, glucose, fructose, galactose, and xylose) and proline in wines, musts, and vinegars can be easily accomplished. The method allows high recovery and satisfies the necessary requirements for accuracy, repeatability, and sensitivity. Values obtained for proline content ranged from 470 to 1190 mg/L in "Aglianico" red wines (mean value, 870 +/- 192 mg/L, n = 21) and from 168 to 286 mg/L in white wines (mean value, 208 +/- 32 mg/L, n = 11). Lower levels were found in musts of red and white grapes, 550 and 87 mg/L, respectively. The lowest content of proline, ca. 10 mg/L, was found both in white and red vinegars.     

Inhibition of human cAMP-phosphodiesterase as a mechanism of the spasmolytic effect of Matricaria recutita L

Mechanisms underlying the spasmolytic activity of chamomile still remain unclear. Inhibition of cAMP- and cGMP-phosphodiesterases (PDE) is one of the mechanisms operated by spasmolytic drugs. In this study, the effect of chamomile on PDE was investigated. Human platelet cAMP-PDE and recombinant PDE5A1 were assayed in the presence of infusions prepared from sifted flowers and capitula. LC-ESI-MS/MS analysis showed different compositions in infusions made with sifted flowers and capitula. Chamomile inhibited cAMP-PDE activity (IC50 = 17.9-40.5 microg/mL), while cGMP-PDE5 was less affected (-15% at 50 microg/mL). Among the individual compounds tested, only flavonoids showed an inhibitory effect (IC50 = 1.3-14.9 microM), contributing to around 39% of the infusion inhibition; other compounds responsible for cAMP-PDE inhibition still remain unknown. Although experimental evidence supporting the use of chamomile for gastrointestinal minor spasms dates back to the fifties, cAMP-PDE inhibition as a likely mechanism underlying the spasmolytic activity is reported for the first time.