Methodology to Improve the Efficiency of Evaluation of the Severity of Angular Leaf Spot in Common Bean

Disease severity assessment by means of a scoring scale, especially for angular leaf spot (Pseudocercospora griseola) in common bean, is hindered in experiments for assessment of progenies and/or breeding lines due to lack of uniformity of occurrence of the pathogens and segregation within progenies. The purpose of this study was to estimate the efficiency of the use of one plant per plot in assessing the severity of angular leaf spot in experiments for assessment of progenies and/or breeding lines in the common bean crop. To that end, two experimental strategies were used – one of them using one plant per plot and another using a standard size plot (SPP) (2–4‐m length rows). The experiments were conducted in the period from November 2011 to May 2012 in the municipalities of Lavras and Lambari, state of Minas Gerais, Brazil. Forty‐one lines from the breeding programme of the Universidade Federal de Lavras (UFLA) and from other research institutions were assessed, which differed in regard to their degree of susceptibility to P. griseola. The lines were assessed in regard to the severity of said disease using a five‐degree diagrammatic scale. In all the one plant per plot experiments, severity scores of angular leaf spot from the beginning of its occurrence, and later in intervals ranging from 7 to 12 days, were obtained. In the experiment with the SPP, assessment was made a few days prior to grain harvest. Estimates of the correlations between severity scores and grain yield (GY) were mostly of small magnitude. There was good coincidence between the lines classified as more resistant or more susceptible to the pathogen under the two conditions.     

Studies on the <Emphasis Type="Italic">Pycnoporus sanguineus</Emphasis> CCT-4518 laccase purified by hydrophobic interaction chromatography

A laccase from Pycnoporus sanguineus was purified by two steps using phenyl-Sepharose columm. A typical procedure provided 54.1-fold purification, with a yield of 8.37%, using syringaldazine as substrate. The molecular weight of the purified laccase was 69 and 68 kDa as estimated by 12% (w/v) SDS-PAGE gel and by gel filtration, respectively. The K _m values for the substrates ABTS, syringaldazine, and guaiacol were 58, 8.3, and 370 μM, respectively. The enzyme’s pH optimum for syringaldazine was 4.2 and optimal activity was 50°C. The enzyme showed to be thermostable because when kept at 50°C for 24 and 48 h it retained 93 and 76% activity. This laccase was inhibited by l-cysteine, β-mercaptoethanol, NaN₃, NaF, and HgCl₂.     

Ultra-deep sequencing reveals the microRNA expression pattern of the human stomach

Background

While microRNAs (miRNAs) play important roles in tissue differentiation and in maintaining basal physiology, little is known about the miRNA expression levels in stomach tissue. Alterations in the miRNA profile can lead to cell deregulation, which can induce neoplasia.

Methodology/Principal Findings

A small RNA library of stomach tissue was sequenced using high-throughput SOLiD sequencing technology. We obtained 261,274 quality reads with perfect matches to the human miRnome, and 42% of known miRNAs were identified. Digital Gene Expression profiling (DGE) was performed based on read abundance and showed that fifteen miRNAs were highly expressed in gastric tissue. Subsequently, the expression of these miRNAs was validated in 10 healthy individuals by RT-PCR showed a significant correlation of 83.97% (P<0.05). Six miRNAs showed a low variable pattern of expression (miR-29b, miR-29c, miR-19b, miR-31, miR-148a, miR-451) and could be considered part of the expression pattern of the healthy gastric tissue.

Conclusions/Significance

This study aimed to validate normal miRNA profiles of human gastric tissue to establish a reference profile for healthy individuals. Determining the regulatory processes acting in the stomach will be important in the fight against gastric cancer, which is the second-leading cause of cancer mortality worldwide.     

Comparative study of the effect of distance on the daily and hourly pollen counts in a city in the south-western Iberian Peninsula

The airborne pollen concentration in the city of Badajoz was measured in two locations 2.9 km apart. The measurements were taken from January to June between 2009 and 2012 using Hirst-type volumetric aerobiological samplers. One sampler was placed at the Faculty of Science (FS) and the other at the Agricultural Engineering School (AES) of the University of Extremadura, Spain, on terraces located 16 and 6 m above ground, respectively. The two sets of hourly and daily pollen concentrations were compared regarding the meteorological parameters and the distribution of local pollen sources. A total of 46 pollen types were counted, with a mean total concentration of 43 pollen grains/m³ in the winter and 336 pollen grains/m³ in the spring. In the winter, pollen grains from (in decreasing order) Cupressaceae, Fraxinus–Phillyrea, Urticaceae spp., Alnus glutinosa and Urtica membranacea types represented 77 % of the total. In the spring, 89 % of the total was represented by pollen grains from (in decreasing order) Quercus, Poaceae, Olea, Pinaceae and Plantago. The FS/AES ratio was 0.57 in the winter and 1.31 in the spring. While a Wilcoxon test applied to the daily total concentration data showed statistically significant differences between the two sites, a correlation study based on the Spearman coefficient showed statistically significant correlations in both the winter and spring. The results were similar when comparing the separate pollen types, except for Urticaceae spp., which showed no statistically significant correlation. The meteorological data studied showed a statistically significant correlation with the daily concentrations. A comparison of the hourly concentration data showed no correspondence with the time of maximum concentration. The local distribution of pollen sources explained some of the differences found between the two sites. Overall, the results indicate that a single aerobiological sampler may be sufficiently representative to register the daily pollen grain data of an urban area of approximately 3 km or greater in diameter, although it underestimates the influence of heterogeneity in the distribution of the local flora.     

Vascular mechanisms involved in angiotensin II-induced venoconstriction in hypertensive rats

To investigate the venoconstrictor effect of angiotensin II (Ang II) in spontaneously hypertensive rats (SHR), we used preparations of mesenteric venular beds and the circular muscle of the portal veins. Vessels were tested with Ang II in the presence or absence of losartan, PD 123319, HOE 140, L-NAME, indomethacin, or celecoxib. In the mesenteric venular bed of SHR, the effect of Ang II (0.1 nmol) was nearly abolished by losartan and enhanced by HOE 140, indomethacin, and celecoxib, while PD123319 and L-NAME had no effect. In portal vein preparations, cumulative-concentration response curves (CCRC) to Ang II (0.1–100 nmol/L) exhibited a lower maximal response (E_max) in SHR compared to Wistar rats. AT₁ receptor expression was similar in the two strains, while AT₂ receptor levels were lower in SHR portal veins when compared to Wistar. In SHR portal veins, losartan shifted the CCRC to Ang II to the right, while indomethacin and HOE 140 increased the E_max to Ang II. PD 123319, celecoxib, and L-NAME had no effect. Taken together, our results suggest that Ang II-induced venoconstriction in SHR is mediated by activation of AT₁ receptors and this effect may be counterbalanced by kinin B₂ receptor and COX metabolites. Furthermore, our data indicate that there are different cellular and molecular mechanisms involved in the regulation of venous tonus of normotensive and hypertensive rats. These differences probably reflect distinct factors that influence arterial and venous bed in hypertension.     

Trophic ecology of three stingrays (Myliobatoidei: Dasyatidae) off the Brazilian north-eastern coast: Habitat use and resource partitioning

Understanding the ecological role of species with overlapping distributions is central to inform ecosystem management. Here we describe the diet, trophic level and habitat use of three sympatric stingrays, Hypanus guttatus, H. marianae and H. berthalutzae, through combined stomach content and stable isotope (δ¹³C and δ¹⁵N) analyses. Our integrated approach revealed that H. guttatus is a mesopredator that feeds on a diverse diet of benthic and epibenthic marine and estuarine organisms, principally bivalve molluscs, Alpheus shrimp and teleost fishes. Isotopic data supported movement of this species between marine and estuarine environments. H. berthalutzae is also a marine generalist feeder, but feeds primarily on teleost fishes and cephalopods, and consequently occupies a higher trophic level. In contrast, H. marianae is a mesopredator specialized on shrimps and polychaetas occurring only in the marine environment and occupying a low niche breadth. While niche overlap occurred, the three stingrays utilized the same prey resources at different rates and occupied distinct trophic niches, potentially limiting competition for resources and promoting coexistence. These combined data demonstrate that these three mesopredators perform different ecological roles in the ecosystems they occupy, limiting functional redundancy.     

Characteristics of Hospitalized Cases of Pertussis in Catalonia and Navarra,Two Regions in the North of Spain

Pertussis causes a large number of cases and hospitalizations in Catalonia and Navarra. We made a study of household cases of pertussis during 2012 and 2013 in order to identify risk factors for hospitalization in pertussis cases. Each primary case reported triggered the study of their contacts. Close contacts at home and people who were in contact for >2 hours during the transmission period of cases were included. The adjusted OR and 95% confidence intervals (CI) was calculated using logistic regression. A total of 1124 pertussis cases were detected, of which 14.9% were hospitalized. Inspiratory whoop (aOR: 1.64; CI: 1.02–2.65), apnoea (aOR: 2.47; CI: 1.51–4.03) and cyanosis (aOR: 15.51; CI: 1.87–128.09) were more common in hospitalized than in outpatient cases. Hospitalization occurred in 8.7% of correctly-vaccinated cases, 41.1% of non-vaccinated cases and 9.4% of partially-vaccinated cases. In conclusion, inspiratory whoop, apnoea and cyanosis were associated factors to hospitalization while vaccination reduced hospitalizations due to pertussis.