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.     

Effects of live prey availability on growth and survival in the early stages of cuttlefish Sepia officinalis (Linnaeus, 1758) life cycle

This investigation examined the effects of live prey availability on growth and survival of Sepia officinalis. Two independent experiments, comprising two feed rations each, were performed, using adequate prey size. In the first experiment, cuttlefish hatchlings were fed live mysids, Paramysis nouvelli [(feed ratio I (fr I)], at 15% body weight per day (bw day^?1) (fr I₁₅) and 30% bw day^?1 (fr I₃₀). In the second experiment, juvenile cuttlefish were fed live Atlantic ditch shrimp, Paleomonetes varians (fr II), under the same experimental design. In both experiments, the final mean weight, feeding rate and instantaneous growth rate were higher when animals were fed feed ratio fr II₁₅ and fr II₃₀ (30% bw day^?1). The results indicate that prey availability influenced weight gain, irrespective of the prey used, during the first 2 months of cuttlefish life cycle. This effect seems to be more noticeable when a certain limit of prey is achieved. For cuttlefish fed fr II, the optimal prey density is thought to be under 2.5 g prey L^?1 (i.e. 14 shrimp L^?1). Results indicate that cuttlefish can withstand prey densities up to 120 mysids L^?1 for cuttlefish up to 3 weeks old and 19 shrimps L^?1 for cuttlefish up to 6 weeks old.     

Effects of binder type and binder addition on the growth of juvenile Palaemonetes varians and Palaemon elegans (Crustacea: Palaemonidae)

This investigation compared the effects of using two binders of no nutritional value, lignosol and agar, and the binding method, microbinding and microcoating, on the growth of juvenile Atlantic ditch shrimp, Palaemonetes varians, and rockpool prawn, Palaemon elegans. Shrimp were fed one basal diet which varied solely in binder type and method of inclusion (Diet A, with lignosol added by microbinding, Diet B, with agar added by microbinding, and Diet C with lignosol added by microcoating) for a period of 45 days. At the end of the experiment significant differences in the final wet weight were found between shrimps fed Diet A and Diet C and between shrimps fed Diet B and Diet C (P < 0.001), but no significant differences were found between Diet A and Diet B (P > 0.05). These results were verified for both species. Significant differences were found between the linear regression slopes of the three tested diets for P. varians (F _3,411 = 3.41; P < 0.04), and for P. elegans (F _3,431 = 10.51; P < 0.0003). Palaemon elegans grew faster than P. varians and at the end of the experiment significant differences were found between the final wet weight of the two species (F _3,842 = 48.17, P < 0.0001). The growth parameters analysed, weight gain (WG), thermal-unit growth coefficient (TGC), and food conversion rate (FCR) were always higher for P. elegans. Results indicate that use of either lignosol or agar, added to the diet by microbinding, is advisable.     

Fractional extraction of compounds from grape seeds by supercritical fluid extraction and analysis for antimicrobial and agrochemical activities

White grape seeds were subjected to sequential supercritical fluid extraction. By increasing the polarity of the supercritical fluid using methanol as a modifier of CO(2), it was possible to fractionate the extracted compounds. Two fractions were obtained; the first, which was obtained with pure CO(2), contained mainly fatty acids, aliphatic aldehydes, and sterols. The second fraction, obtained with methanol-modified CO(2), had phenolic compounds, mainly catechin, epicatechin, and gallic acid. The fractions were bioassayed. Antimicrobial activities were checked on human pathogens, and a high degree of activity was obtained with the lipophilic fraction. Agrochemical activities on phytopathogenic fungi and activities on the etiolated wheat coleoptile bioassay were also checked. The more polar fraction was active in the latter bioassay.     

Residue level, persistence, and storage performance of citrus fruit treated with fludioxonil

The potential of postharvest dip treatments with fludioxonil (FLU) (a synthetic analogue of the bacterial metabolite of pyrrolnitrin), in controlling postharvest decay caused by Penicillium digitatum and Penicillium italicum of citrus fruit was investigated in comparison with the conventional fungicide imazalil (IMZ). The ultrastructural changes of fruit epicuticular wax was investigated as a function of water dip temperature, and the possible role of these changes was related to residue accumulation under FLU treatment. Residues retained by fruit were determined as a function of fungicide concentration, dip temperature, and fruit storage conditions. Scanning electron microscopy analysis revealed that fruit dipping in water at 30 or 40 degrees C did not cause differences in cuticular wax's ultrastructure in comparison to control fruit, while treatments at 50, 55, or 60 degrees C caused the disappearance of wax platelets, resulting in relatively homogeneous skin surface, due to partial "melting" of epicuticular wax. Residues of FLU in fruit treated at 20 or 50 degrees C were significantly correlated with the doses of fungicide applied. When equal amounts of fungicide were employed, the residue concentrations were notably higher (from 2.6- to 4-fold) in fruit treated at 50 degrees C than in fruit treated at 20 degrees C. The dissipation rate of FLU in "Salustiana" and "Tarocco" oranges was lower in fruit subjected to treatment at 50 degrees C. The minimal FLU concentration for almost complete decay control in artificially wounded fruit during 7-d storage at 20 degrees C was 400 mg/L active ingredient (ai) in fruit treated at 20 degrees C and 100 mg/L ai in fruit treated at 50 degrees C. Results on nonwounded Tarocco oranges subjected to 3 weeks of simulated quarantine conditions at 1 degrees C, plus 6 weeks of standard storage at 8 degrees C and an additional two weeks of simulated marketing period (SMP) at 20 degrees C revealed that almost complete decay control with FLU applications of 100 mg/L at 50 degrees C and 400 mg/L at 20 degrees C resulted in ca. 0.8 mg/kg FLU fruit residues, in agreement with results on wounded citrus fruit. When equal concentrations and temperatures were applied, FLU treatments were as effective as IMZ. In vitro trials showed a low sensitivity to FLU against P. digitatum and P. italicum isolates. MIC values for the complete inhibition of mycelium growth were >or=100 microg/mL, while ED(50) values ranged from 0.1 to 1 microg/mL for P. digitatum and from 1 to >100 microg/mL for P. italicum. The latter result suggests that care should be taken to avoid exclusive application of FLU in a sustainable program for management of fruit decay. However, integrating fungicide application and hot water dip may reduce the possibility of selecting fungicide-resistant populations of the pathogen, by increasing the effectiveness of the treatment.     

Proteomic profiles during adventitious rooting of Eucalyptus species relevant to the cellulose industry

New Forests - Adventitious rooting (AR) is an obligatory step for vegetative propagation of commercial woody species. Paper industries have interest in Eucalyptus globulus Labill and its hybrids...     

Challenges and opportunities in polymer technology applied to veterinary medicine

An important frontier in the administration of therapeutic drugs to veterinary species is the use of different polymers as drug delivery platforms. The usefulness of polymers as platforms for the administration of pharmaceutical and agricultural agents has been clearly recognized in the recent decades. The chemical versatility of polymers and the wide range of developed controlled‐release strategies enhance the possibilities for the formulation of active molecules. In particular, the veterinary area offers opportunities for the development of novel controlled‐release drug delivery technologies adapted to livestock or companion animal health needs. In some cases, it also allows to improve profitability in meat production or to meet the safety criteria related to drug residues. A number of factors affect the selection of polymers and subsequent properties of the controlled‐release drug delivery system. However, their selection also dictates the release kinetics of the drug from the delivery system. Such choices are therefore crucial as they affect the success and potential of the delivery system for achieving the therapeutic goals of the veterinarian. It is the intention of this review to give an overview of the most relevant polymers, which are used or have been tested as drug delivery release rate modifiers in the veterinary field. The article highlights some recent developments focusing on their advantages and applications and analyzes the future direction of the scientific and technological advancements in this area.     

Landscape-scale modelling of agroforestry ecosystems services in Swiss orchards: a methodological approach

Context

Agroforestry systems in temperate Europe are known to provide both, provisioning and regulating ecosystem services (ES). Yet, it is poorly understood how these systems affect ES provision at a landscape scale in contrast to agricultural practises.

Objectives

This study aimed at developing a novel, spatially explicit model to assess and quantify bundles of provisioning and regulating ES provided by landscapes with and without agroforestry systems and to test the hypothesis that agroforestry landscapes provide higher amounts of regulating ES than landscapes dominated by monocropping.

Methods

Focussing on ES that are relevant for agroforestry and agricultural practices, we selected six provisioning and regulating ES—“biomass production”, “groundwater recharge”, “nutrient retention”, “soil preservation”, “carbon storage”, “habitat and gene pool protection”. Algorithms for quantifying these services were identified, tested, adapted, and applied in a traditional cherry orchard landscape in Switzerland, as a case study. Eight landscape test sites of 1 km?×?1 km, four dominated by agroforestry and four dominated by agriculture, were mapped and used as baseline for the model.

Results

We found that the provisioning ES, namely the annual biomass yield, was higher in landscape test sites with agriculture, while the regulating ES were better represented in landscape test sites with agroforestry. The differences were found to be statistically significant for the indicators annual biomass yield, groundwater recharge rate, nitrate leaching, annual carbon sequestration, flowering resources, and share of semi-natural habitats.

Conclusions

This approach provides an example for spatially explicit quantification of provisioning and regulating ES and is suitable for comparing different land use scenarii at landscape scale.

     

The acute phase response induced by Escherichia coli lipopolysaccharide modifies the pharmacokinetics and metabolism of florfenicol in rabbits

The aim of this study was to determine the effect of Escherichia coli lipopolysaccharide (LPS)‐induced acute phase response (APR) on the pharmaco‐kinetics and biotransformation of florfenicol (FFC) in rabbits. Six rabbits (3.0 ± 0.08 kg body weight (bw)) were distributed through a crossover design with 4 weeks of washout period. Pairs of rabbits similar in bw and sex were assigned to experimental groups: Group 1 (LPS) was treated with three intravenous doses of 1 μg/kg bw of E. coli LPS at intervals of 6 h, and Group 2 (control) was treated with an equivalent volume of saline solution (SS) at the same intervals and frequency of Group 1. At 24 h after the first injection of LPS or SS, an intravenous bolus of 20 mg/kg bw of FFC was administered. Blood samples were collected from the auricular vein before drug administration and at different times between 0.05 and 24.0 h after treatment. FFC and florfenicol‐amine (FFC‐a) were extracted from the plasma, and their concentrations were determined by high‐performance liquid chromatography. A noncompartmental pharmacokinetic model was used for data analysis, and data were compared using the paired Student t‐test. The mean values of AUC_0–∞ in the endotoxaemic rabbits (26.3 ± 2.7 μg·h/mL) were significantly higher (P < 0.05) than values observed in healthy rabbits (17.2 ± 0.97 μg·h/mL). The total mean plasma clearance (CL_T) decreased from 1228 ± 107.5 mL·h/kg in the control group to 806.4 ± 91.4 mL·h/kg in the LPS‐treated rabbits. A significant increase (P < 0.05) in the half‐life of elimination was observed in the endotoxaemic rabbits (5.59 ± 1.14 h) compared to the values observed in healthy animals (3.44 ± 0.57 h). In conclusion, the administration of repeated doses of 1 μg/kg E. coli LPS induced an APR in rabbits, producing significant modifications in plasma concentrations of FFC leading to increases in the AUC, terminal half‐life and mean residence time (MRT), but a significant decrease in CL_T of the drug. As a consequence of the APR induced by LPS, there was a reduction in the metabolic conversion of FFC to their metabolite FFC‐a in the liver, suggesting that the mediators released during the APR induced significant inhibitory effects on the hepatic drug‐metabolizing enzymes.