A New Bioassay Platform Design for the Discovery of Small Molecules with Anticancer Immunotherapeutic Activity

Immunotherapy takes advantage of the immune system to prevent, control, and eliminate neoplastic cells. The research in the field has already led to major breakthroughs to treat cancer. In this work, we describe a platform that integrates in vitro bioassays to test the immune response and direct antitumor effects for the preclinical discovery of anticancer candidates. The platform relies on the use of dendritic cells that are professional antigen-presenting cells (APC) able to activate T cells and trigger a primary adaptive immune response. The experimental procedure is based on two phenotypic assays for the selection of chemical leads by both a panel of nine tumor cell lines and growth factor-dependent immature mouse dendritic cells (D1). The positive hits are then validated by a secondary test on human monocyte-derived dendritic cells (MoDCs). The aim of this approach is the selection of potential immunotherapeutic small molecules from natural extracts or chemical libraries.     

Artenspektrum und Befallshäufigkeit von Fusarium spp. in Bt- und konventionellem Mais im Maiszünsler-Befallsgebiet Oderbruch

The European corn borer (Ostrinia nubilalis) is one of the most important pests of maize (Zea mays). Injuries to the plants caused by the larvae of the European corn borer may represent entrance gates for fungal-spores. The cultivation of Bacillus thuringiensis maize (Bt-maize) is one possibility to reduce infestation by the European corn borer. The aim of the present project was to determine and to compare the number of species and the frequency of Fusarium spp. infestation in Bt-maize (cry1Ab) and conventional maize. In 2003, we analysed the Fusarium spp. infestation of samples of chaffed Bt-maize and its isogenic variety on two experimental fields in the Oderbruch region (Germany), an European corn borer infested area. The conventional variety on the first of the experimental fields (previous crop wheat and forking cultivation) showed a small infestation (16%) of Ostrinia nubilalis while in the conventional variety on the second field (previous crop maize and not forking cultivation) the infestation of the European corn borer was almost three times higher (47%). In the conventional variety on both of the experimental fields we found a high Fusarium spp. infestation (70%). Especially species of the section Liseola dominates, among them: F. subglutinans, F. proliferatum und F. verticillioides. The Fusarium infestation in the samples of Bt-maize from the field with previous crop wheat and forking cultivation was just as high as in the conventional variety (70%). The infestation of Fusarium spp. in the samples of Bt-maize from the field with previous crop maize and forking cultivation was more than 20?% lower.     

Multi‐temporal Digital Photogrammetric Analysis for Quantitative Assessment of Soil Erosion Rates in the Landola Catchment of the Upper Orcia Valley (Tuscany,Italy)

This paper focuses on the analysis of the slope and catchment erosion dynamics in a typical Mediterranean context and its sensitivity to recent climatic and socio‐economic changes. The main objective of the present study is to test the reliability of digital photogrammetric analysis to evaluate the time and space evolution of erosion processes mainly triggered by surface running waters and landslides for about the last 60 years. The selected test area is the Landola catchment, a minor tributary of the Upper Orcia River Valley (Southern Tuscany). The Upper Orcia valley is a key site for the comprehension of denudation processes typically acting in Mediterranean badland areas thanks to the following: (i) the availability of long‐lasting erosion monitoring datasets (20 years' long direct measurements at erosion ‘hot spots’); (ii) its representativeness as a sub‐humid Mediterranean badland area; and (iii) the rapidity of development of erosion processes, which makes it suitable as an open‐air laboratory for the study of badland dynamics. The outputs of this work highlight that the application of high‐resolution digital photogrammetric methodologies can represent a powerful and low‐cost tool to evaluate rates and spatial–temporal distribution of denudation processes, as confirmed by the validation through field point monitoring in areas close to the study area. The results obtained for the study area confirm that high erosion rates are a persistent environmental problem for the Upper Orcia Valley, which is not solved up to now, despite various land conservation interventions. The performed multi‐temporal analysis shows a slight decrease in the average water erosion rate during the last 60 years and a parallel increase in the frequency of mass wasting events. These variations are most likely related to a complex interplay between land use changes that have affected the study area during the studied period, revealing a very delicate equilibrium between farming activities and erosion processes. Copyright © 2014 John Wiley & Sons, Ltd.     

Preparation and promotion of fruit growth in kiwifruit of fluorinated N-phenyl-N'-1,2,3-thiadiazol-5-yl ureas

Seventeen phenyl-fluorinated analogues of thidiazuron [N-phenyl-N'-(1,2,3-thiadiazol-5-yl)urea, TDZ] have been prepared and characterized. The effects of each fluorinated urea on growth and quality of kiwifruits (Actinidia deliciosa) were evaluated by comparison with untreated (control) and TDZ-treated fruits. The results obtained showed a clear dependence of the growth-promoting activity of these fluorinated ureas on the pattern and degree of fluorine substitution in the phenyl ring. The most effective for promoting fruit growth was N-(2,3,5,6-tetrafluorophenyl)-N'-(1',2',3'-thiadiazol-5'-yl)urea at 25 ppm (at harvest, treated fruits were 58% heavier than untreated ones) followed by N-(3,5-difluorophenyl)-N'-(1',2',3'-thiadiazol-5'-yl)urea at 10 ppm (50%). Comparatively, TDZ-treated fruits were 31% (10 ppm) and 38% (25 ppm) heavier than untreated ones. The results also indicate that the effects of the more active phenyl-fluorinated ureas on some standard quality parameters of fruits, for example, percent of fruit dry matter content, soluble solids contents, total titratable acids, shape, and internal structure, are similar to those of TDZ. Quantitative structure-activity relationships have been derived for the fruit growth promoting activity of the phenyl-fluorinated analogues of TDZ.     

Evaluation of the flammability of gorse (<Emphasis Type="Italic">Ulex europaeus</Emphasis> L.) managed by prescribed burning

• Introduction  

The abandonment of rural areas has led to an increase of the fire-prone European gorse (Ulex europaeus L.) communities in some regions, where prescribed burning is a technique applied to control them. Understanding flammability changes after treatments is crucial for the sustainable use of fire.     

Tissue distribution of Red Spotted Grouper Nervous Necrosis Virus (RGNNV) genome in experimentally infected juvenile European seabass (Dicentrarchus labrax)

The distribution of viral genome in the tissues of juvenile European seabass (Dicentrarchus labrax) during the course of a Red Spotted Grouper Nervous Necrosis Virus (RGNNV) infection has not yet been described. The present study addresses this and indicates which target organs may be involved in viral replication. This information should enable more accurate detection of virus in asymptomatic carriers, and in turn help to control the spread of the disease. The aim of this study was to examine the pattern of expression of viral genomic segments RNA1 and RNA2, using two absolute real-time PCRs (RT-qPCR), over the course of a RGNNV infection after administering the virus by intramuscular injection. In situ hybridization was also used to locate the RNA2 viral segment in different organs throughout the infection. The experimental challenge provoked an acute form of viral nervous necrosis (VNN), with a resulting cumulative mortality of 37%. The RT-qPCRs designed allowed the detection of both genomic segments in all the organs tested (nervous and non-nervous tissues) at all sampling times examined. The highest viral RNA copy number was found in eyes, although viral replication appeared to begin in the brain. Viral replication was also recorded in pooled internal organs and in caudal fin. However, the increase in the viral RNA copy number in these organs did not result in an increased viral titre, which may indicate that a productive infection does not take place in non-nervous tissues, possibly due to a failure in a viral post-replication step.     

Characterization of Collagen from Different Discarded Fish Species of the West Coast of the Iberian Peninsula

ABSTRACT

Skin collagen of six discarded fish species was analyzed and compared. Acid soluble collagen (ASC) was extracted; a characteristic sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) profile for type I collagen was obtained, except for Chimaera mostrosa. Contents of collagen calculated from HPro (31.85% average) were higher than those determined from ASC extracts (17.75% average), with Galeus spp. being the species with the higher percentage. Amino acid analysis revealed the typical composition of collagen, with very few differences among species. Specific profiles were obtained after protease digestion. Denaturation temperature of ASC correlated well with imino and hydroxyproline contents.

Results demonstrate the feasibility of using the obtained collagens in different industrial applications.     

Changes in the free amino acid contents of honeys during storage at ambient temperature

This study was carried out to establish the changes in the free amino acid contents of floral honeys, honeydew honeys, and blend honeys during storage at room temperature and to test the capacity of the amino acids to distinguish the origin of the honeys after storage. For this purpose, 54 artisanal honeys (39 floral, 5 honeydew, and 10 blend) were studied. Samples were taken from recently collected honeys and at 3, 6, 9, 12, 16, 20, and 24 months after harvesting. The contents of most of the free amino acids were found to decrease with storage time, with the greatest reduction observed in the first 9 months. The contents of the amino acids aspartic acid, beta-alanine, and proline increased in the first few months after storage, reaching maximum values at 6 months, suggesting the possible existence of enzymatic activities. The application of stepwise discriminant analysis to the free amino acid content data demonstrated that the contents of the amino acids valine, beta-alanine, gamma-aminobutyric acid, serine, isoleucine, alpha-alanine, ornithine, and glutamine correctly assigned 87% of honeys to their group of origin: floral, honeydew, or blend.     

The role of private conservation areas in biodiversity representation and target achievement within the Little Karoo region, South Africa

It is becoming increasingly difficult to manage and expand statutory conservation areas (i.e., parks and formally protected areas). Therefore, alternative opportunities for land conservation merit closer attention. This paper examines the extent to which privately owned conservation areas contribute to biodiversity representation. Gap analyses were performed for a large semi-arid region in South Africa with a comprehensive database of private conservation areas. The distribution of private conservation areas was compared to statutory conservation areas using several landscape characteristics: biome and vegetation variant, elevation class, ecological process area, total area, and threat status (endangerment). Conservation target achievement for the vegetation variants was also assessed, as was the degree to which private conservation areas complemented statutory conservation areas by representing different landscape characteristics. The number of targets achieved nearly tripled if private conservation areas were considered in addition to statutory conservation areas. Further, private conservation areas significantly complemented statutory conservation areas in the types of biomes, elevation classes, and threat status classes conserved. Private conservation areas were especially important in conserving lower elevation habitat, and by association, endangered vegetation. This particular relationship is expected to be common worldwide. Our results indicate that private lands conservation deserves an increased allocation of resources for both research and implementation.     

Laboratory characterization of firebrands involved in spot fires

• Introduction  

Wildfires are considered the most important disturbance in the Mediterranean Basin, and some are propagated over long distances due to lift-off and ignition of firebrands.