Involvement of neuronal nitric oxide synthase (nNOS) in the regulation of migrating motor complex (MMC) in sheep

The objectives of this study were to evaluate the role of nitric oxide (NO) synthase isoforms (nNOS, eNOS, and iNOS) in the regulation of the migrating motor complex (MMC) in sheep using electromyography and their expression in the gastrointestinal (GI) tract by Western blot (WB) and immunohistochemistry. Intravenous administration of L-NAME or the nNOS inhibitor 7-nitroindazole (7-NI) decreased the MMC interval. Myoelectric activity of intestinal phase II was increased, whereas antral activity was reduced. These effects were blocked by L-arginine. Inhibitors of either iNOS (aminoguanidine and S-methylisothiourea) or eNOS (L-NIO) were ineffective. The NO donor sodium nitroprusside decreased GI myoelectric activity, inhibited the MMC pattern, and prevented the effects induced by L-NAME and 7-NI in the intestine. Intracerebroventricular administration of these agents did not modify GI motility. In the rumen, abomasal antrum, duodenum, and jejunum, WB showed three bands at about 155, 145, and 135kDa corresponding to nNOS, and a 140-kDa band (eNOS); however iNOS was not detected. Positive nNOS immunostaining was observed in neurons of the myenteric and submucous plexus of all GI tissues, while eNOS was found in the endothelial cells, ruminal and intestinal epithelium, as well as in some enteric neurons and in endocrine-like cells of the duodenal Brunner's glands. In contrast, only weak iNOS immunoreactivity was found in ruminal epithelium. Taken together, our results suggest that NO, synthesized at a peripheral level by nNOS, is tonically inhibiting the MMC pattern and intestinal motility in sheep.     

The Utility of Flow Cytometry for the Diagnosis of Cranial Mediastinal Malignancy in Canine Patients

Introduction:  The most common neoplasms located in the anterior mediastinum in the canine are thymoma and lymphoma. Distinguishing between the two is a diagnostic challenge. Treatment and prognosis for these diseases differs significantly. Thymomas contain a population of normally developing T cells. The majority of these T cells exhibit an immature phenotype, characterized by co‐expression of CD4 and CD8. This phenotype is rarely seen on neoplastic lymphocytes. The purpose of this study was to determine if analysis by flow cytometry could discriminate thymoma from lymphoma based on these cell surface markers.
Methods:  Fine needle aspirates were obtained from ten canine patients with mediastinal masses. Cells were analyzed by flow cytometry using a panel of T and B cell markers.
Results:  Six cases with 10% or greater CD4 + CD8+ cells were diagnosed with thymoma and were confirmed by histopathology. Four cases had fewer than 5% CD4 + CD8+ cells, having lymphocytes expressing CD4 only (3 cases) or CD21, a B cell marker(1 case). These were confirmed as lymphoma by cytology and/or a clonality assay. The sensitivity and specificity of this assay when used in the diagnostic work‐up for suspected thymoma was 100%.
Conclusion:  Flow cytometry may provide important, complementary information in the diagnostic work‐up of the canine patient with a mediastinal mass.     

The utility of the ‘Arable Weeds and Management in Europe’ database: Challenges and opportunities of combining weed survey data at a European scale

Over the last 30 years, many studies have surveyed weed vegetation on arable land. The ‘Arable Weeds and Management in Europe’ (AWME) database is a collection of 36 of these surveys and the associated management data. Here, we review the challenges associated with combining disparate datasets and explore some of the opportunities for future research that present themselves thanks to the AWME database. We present three case studies repeating previously published national scale analyses with data from a larger spatial extent. The case studies, originally done in France, Germany and the UK, explore various aspects of weed ecology (community composition, management and environmental effects and within-field distributions) and use a range of statistical techniques (canonical correspondence analysis, redundancy analysis and generalised linear mixed models) to demonstrate the utility and versatility of the AWME database. We demonstrate that (i) the standardisation of abundance data to a common measure, before the analysis of the combined dataset, has little impact on the outcome of the analyses, (ii) the increased extent of environmental or management gradients allows for greater confidence in conclusions and (iii) the main conclusions of analyses done at different spatial scales remain consistent. These case studies demonstrate the utility of a Europe-wide weed survey database, for clarifying or extending results obtained from studies at smaller scales. This Europe-wide data collection offers many more opportunities for analysis that could not be addressed in smaller datasets; including questions about the effects of climate change, macro-ecological and biogeographical issues related to weed diversity as well as the dominance or rarity of specific weeds in Europe.     

Growth and mortality of larval anchoveta Engraulis ringens,in northern Chile during winter and their relationship with coastal hydrographic conditions

Larval growth, age, growth effect and instantaneous mortality were estimated in anchoveta, Engraulis ringens, collected biweekly during the austral winter of 2014 in nearshore waters off Bay of Antofagasta (23°41′W–70°30′S), northern Chile. Through measuring standard length (SL) and sagitta microstructure analysis, it was estimated that the growth rate of E. ringens larvae decreased from June (0.85 mm day^?1) to August (0.50 mm day^?1). However, the water temperature was homogeneous during the sampling dates (14.6, 15.2, 14.4, and 14.6°C), suggesting that the decelerating larval growth was not linked to changes in sea temperature. Additionally, larvae with slow growth have larger otoliths compared with conspecifics with fast growth (growth effect). Larval mortality rates tended to decrease until the middle of July (0.18 per day) but increased to 0.25 per day in early August, which coincided with lower food availability (i.e., chlorophyll‐a, 2.7–5.6 mg m^?3) and a high occurrence of smaller larvae (1.58–11.5 mm). Partial least squares analysis indicates low covariance between the biological and oceanographic variables (PLS: 11.71%), suggesting that other factors, such as parental effects, may explain the abrupt decrease in the larval growth rates.     

Anchoveta Engraulis ringens along the Chilean coast: Management units,demographic units and water masses: Insights from multiple otolith‐based approaches

Three management units (MUs) are used for stock assessment and fishery regulations of anchoveta Engraulis ringens in Chilean waters (>65% of E. ringens latitudinal range): MU‐I [18–25)°S, MU‐II [25–32)°S and MU‐III [32–42)°S. To evaluate whether these three MUs correspond to separate demographic units (DUs), as well as to estimate mixing rates and spatial distribution of these DUs, we combined three types of otolith natural markers: isotopic signatures (δ¹³C and δ¹⁸O), elemental compositions (Na, Mg, Mn, Sr and Ba) and microstructural indexes. All otolith markers were determined in nuclear and marginal regions of juveniles and adults from cohorts 2012 and 2015. Differences in core region markers indicated spatial segregation between three environmentally distinct nursery areas, probably related to Subtropical Water (MU‐I), Subantarctic Water (MU‐III) and the Subtropical Convergence (MU‐II). Comparison of core and marginal regions, from juvenile and/or adult fish, suggested that adult fish remained nearby nursery areas, separated from fish nursed in other MUs. Nonetheless, ontogenetic migrations from warmer (offshore) nursery habitats to cooler (deeper/more coastal) feeding habitats occurred within MUs. In summary, our results support the consideration of the three MUs as three separate DUs. Estimated mixing rates indicated MU‐II received contributions of 31% from MU‐I and 3% from MU‐III, while complete segregation existed between MU‐I and MU‐III. Such mixing rates between MU‐I and MU‐III seem large enough to justify its further consideration by stock assessment models and management procedures.     

Diagnosis of mediastinal masses in dogs by flow cytometry

BACKGROUND: Biopsy of mediastinal masses can be invasive, but the procedure may be necessary if cytology of a mass aspirate is inconclusive. The 2 most common mediastinal masses, lymphoma and thymoma, may both be comprised of small lymphocytes. We investigated the ability of flow cytometry to distinguish between these 2 neoplasms. HYPOTHESIS: Flow cytometry of mediastinal mass aspirates may provide a definitive diagnosis of thymoma or lymphoma, reducing the need for biopsy. ANIMALS: Dogs with mediastinal masses presenting to the Veterinary Teaching Hospital/Animal Cancer Center were included in the study. METHODS: Aspirates obtained over 2 years that met the inclusion criteria (i.e. sufficient viable cells and a definitive diagnosis by means other than flow cytometry) were analyzed by flow cytometry to determine the percentage of cells expressing B- and T-cell markers, and co-expressing CD4 and CD8. RESULTS: All cases of thymoma (n = 6) consisted of > or = 10% lymphocytes coexpressing CD4 and CD8, a phenotype that is characteristic of thymocytes, whereas 6 of 7 lymphomas contained <2% CD4+CD8+ lymphocytes. The CD4+CD8+ lymphoma could be readily distinguished flow cytometrically from thymoma by light scatter properties. The phenotypes of the remaining lymphomas were CD4+ T cell (4), CD34+ (1) and B cell (1). CONCLUSIONS: Our studies demonstrate that flow cytometry is a useful tool for discriminating mediastinal masses. Lymphocyte-rich mediastinal masses could be unambiguously identified by flow cytometry in 13/13 cases.     

Tillage effects on soil aggregation and soil organic carbon profile distribution under Mediterranean semi‐arid conditions

In rainfed semi‐arid agroecosystems, soil organic carbon (SOC) may increase with the adoption of alternative tillage systems (e.g. no‐tillage, NT). This study evaluated the effect of two tillage systems (conventional tillage, CT vs. NT) on total SOC content, SOC concentration, water stable aggregate‐size distribution and aggregate carbon concentration from 0 to 40 cm soil depth. Three tillage experiments were chosen, all located in northeast Spain and using contrasting tillage types but with different lengths of time since their establishment (20, 17, and 1‐yr). In the two fields with mouldboard ploughing as CT, NT sequestered more SOC in the 0–5 cm layer compared with CT. However, despite there being no significant differences, SOC tended to accumulate under CT compared with NT in the 20–30 and 30–40 cm depths in the AG‐17 field with 25–50% higher SOC content in CT compared with NT. Greater amounts of large and small macroaggregates under NT compared with CT were measured at 0–5 cm depth in AG‐17 and at 5–10 cm in both AG‐1 and AG‐17. Differences in macroaggregate C concentration between tillage treatments were only found in the AG‐17 field at the soil surface with 19.5 and 11.6 g C/kg macroaggregates in NT and CT, respectively. After 17 yr of experiment, CT with mouldboard ploughing resulted in a greater total SOC concentration and macroaggregate C concentration below 20 cm depth, but similar macroaggregate content compared with NT. This study emphasizes the need for adopting whole‐soil profile approaches when studying the suitability of NT versus CT for SOC sequestration and CO₂ offsetting.     

Epigallocatechin‐3‐Gallate (EGCG) Reduces Rotenone Effect on Stallion Sperm–Zona Pellucida Heterologous Binding

Stallion spermatozoa are highly dependent on oxidative phosphorylation for ATP production to achieve normal sperm function and to fuel the motility. The aim of this study was to evaluate the response of equine sperm under capacitating conditions to the inhibition of mitochondrial complex I by rotenone and to test whether epigallocatechin‐3‐gallate (EGCG), a natural polyphenol component of green tea, could counteract this effect. After 2‐h incubation of stallion spermatozoa in modified Tyrode's medium, rotenone (100 nm , 500 nm and 5 μm ) and EGCG (10, 20 and 60 μm ), alone or in combination, did not induce any significant difference on the percentage of viable cells, live sperm with active mitochondria and spermatozoa with intact acrosome. The inhibition of complex I of mitochondrial respiratory chain of stallion sperm with rotenone exerted a negative effect on heterologous ZP binding ability. EGCG at the concentrations of 10 and 20 μm (but not of 60 μm ) induced a significant increase in the number of sperm bound to the ZP compared with that for control. Moreover, when stallion sperm were treated with rotenone 100 nm , the presence of EGCG at all the concentrations tested (10, 20 and 60 μm ) significantly increased the number of sperm bound to the ZP up to control levels, suggesting that this green tea polyphenol is able to reduce the toxicity of rotenone.     

Impact of high pressure and pulsed electric fields on bioactive compounds and antioxidant activity of orange juice in comparison with traditional thermal processing

Bioactive compounds (vitamin C, carotenoids, and flavanones) and DPPH* radical scavenging capacity (RSC) were measured in orange juice (OJ) subjected to different technologies. High pressure (HP) (400 MPa/40 degrees C/1 min), pulsed electric fields (PEF) (35 kVcm(-1)/750 micros), low pasteurization (LPT) (70 degrees C/30 s), high pasteurization (HPT) (90 degrees C/1 min), HPT plus freezing (HPT+F) (-38 degrees C/15 min), and freezing (F) were studied. Among the treatments assayed, even though the losses in total vitamin C were < 9%, treatments with the higher temperatures tended to show the higher decrease in the content of both forms of vitamin C. HP treatment led to an increased (P < 0.05) carotenoid release (53.88%) and vitamin A value (38.74%). PEF treatment did not modify individual or total carotenoids content. Traditional thermal treatments did not exert any effect on total carotenoid content or vitamin A value. With regard to individual carotenoid extraction, HPT and HPT+F led to different releases of carotenoids. With respect to flavanones, HP treatment led to increased (P < 0.05) naringenin (20.16%) and hesperetin (39.88%) contents, whereas PEF treatment did not modify flavanone content. In general, pasteurization and freezing process led to a diminished (P < 0.05) naringenin content (16.04%), with no modification in hesperetin. HP and PEF treatments did not modify DPPH* RSC. In the case of traditional thermal technologies, HPT treatment showed a decrease (P < 0.05) in RSC (6.56%), whereas LPT, HPT+F, and F treatments did not modify RSC. Vitamin C modulated RSC, in terms of antioxidant concentration (EC50) and kinetics (AE = 1/EC50TEC50), in the treated and untreated OJ. In summary, HP and PEF technologies were more effective than HPT treatment in preserving bioactive compounds and RSC of freshly squeezed orange juice.     

The influence of salinity on the vegetative growth,osmolytes and chloride concentration of four halophytic species

The aim of this work was to study the influence of salinity of the fertigation solution on the vegetative growth, as well as on the osmolytes and chloride concentration of four halophytic species. Results show that the increase of salinity caused the reduction of plant fresh and dry matter in Asteriscus maritimus, while plant dry weight was unaffected by the salinity in Crithmum maritimum, Halimione. portulacoides and Limonium cossonianum. Salinity enhanced root growth of H. portulacoides. Salt-induced succulence was detected in A. maritimus. The translocation of Cl^– to the leaves is an important factor responsible for salt tolerance of A. maritimus and H. portulacoides. However, C. maritimum and L. cossonianum restricted the uptake of Cl^– and excrete salts through the leaves. Crithmum maritimum and H. portulacoides accumulated proline and soluble sugars in leaves which acts as osmoprotectant. Among the species studied, H. portulacoides has the greatest Cl^– phytoextraction efficiency.