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.     

Effect of 1-MCP treatment and N2O MAP on physiological and quality changes of fresh-cut pineapple

The effects of both 1-MCP treatment of pineapples and packaging of their fresh-cut products with an alternative modified atmosphere (MA: 86.13 kPa N₂O, 10.13 kPa O₂ and 5.07 kPa CO₂) on physiological and quality changes of these minimally processed products were investigated. Fresh-cut fruit treated or not with 1-MCP were packed in Air or in MA and were stored at 4 °C for 10 d. The following parameters were monitored during storage: ripening index; O₂, CO₂ and C₂H₄ in the package headspace; firmness and colour. Microbial spoilage of MP pineapple samples was also investigated and a mathematical model based on the Zwietering modified Gompertz equation was used to obtain growth parameters of mesophilic bacteria, yeasts and moulds.The results showed that 1-MCP treatment and MAP in a N₂O enriched atmosphere had a positive combined effect on the inhibition of respiration and ethylene production of fresh-cut pineapple and on its softening delay, confirming previous findings about 1-MCP and N₂O preservative effects on fresh-cut fruit quality. This combined effect was not extended to the ripening index and colour maintenance, as MAP at 86.13 kPa of N₂O did not add any benefit to that of the 1-MCP treatment. From a microbiological point of view, N₂O MAP extended the shelf-life of the products of 3–4 d by increasing the lag phase of microbial growth.     

Culture of Macrobrachium rosenbergii (De Man 1879) in experimental cages in a freshwater eutrophic lake at different stocking densities

Macrobrachium rosenbergii (de Man 1879) juveniles (0.4 g) were cultured in experimental cages (L × W × H: 2.5 × 1 × 1 m) in Laguna de Bay, the largest lake in the Philippines. The following stocking densities at four replicates each were used: 15, 30, 60 and 90 prawns m⁻² of cage bottom. The mean sizes at harvest after 5 months of culture ranged from 14.3 g for the highest stocking density to 26.3 g for the lowest. The mean size at harvest, daily growth rate and size class distribution were significantly influenced by stocking density, with those at the lowest stocking density showing significantly better growth and overall proportion of larger prawns. Heterogeneous individual growth (HIG) was fairly evident in all treatments. The percentage of blue‐clawed males was not influenced by treatment but the mean weight was significantly higher in the lower stocking densities. Both the percentage and mean weight of berried females were significantly higher in the lowest stocking density. Survival was the highest in the lower stocking densities (55.3%, 54.0%, 52.7% and 36.9% for 15, 30, 60 and 90 prawns m⁻² respectively). Feed conversion ratio (FCR) improved with decreasing stocking density, ranging from 2.1 to 3. As expected, yield per cropping increased with stocking density and ranged from 450 to 1089 g m⁻² yr⁻¹ of actual cage area. Production values obtained in the cage cultured M. rosenbergii were comparable to or even higher than those reported from pond culture, given that the stocking densities used in this study were generally higher than in ponds. The results show that the farming of M. rosenbergii in cages in lakes is a viable alternative to pond culture and has the potential of improve aquaculture production in lakeshore fish farming communities.     

Sulfometuron resistance as a genetic marker for yeast populations in wine fermentations

Winemaking with selected yeasts requires simple and cheap techniques to monitor the yeast population dynamics. We obtained new sulfometuron (smr) resistant mutants, easy to detect by replica-plate assay, from selected wine yeasts. The mutations were dominant and were located at the ilv2 locus that encodes for acetolactate synthase enzyme. The mutants were genetically stable and maintained the killer phenotype of the parent yeast strain. They were genetically improved by elimination of recessive growth-retarding alleles followed by spore clone selection according to the must fermentation kinetics and the organoleptic quality of the wine. Some mutants were tested in industrial winemaking and were easily monitored during must fermentation using a simple plate assay. They accounted for more than 95% of the total yeasts in the must, and the resulting wine had as good a quality as those made with standard commercial wine yeasts.     

The Representative Day Technique in the Study of Photochemical Smog Pollution in Falconara Industrial Area

The issue of increasing tropospheric Ozone and, more in general, of photochemical smog is of growing in importance. It is, however, a problem of extreme complexity, involving both atmospheric diffusion processes and complex chemical reactions. We present a study carried out to examine the evolution of the Ozone concentration fields and the effects of meteorological conditions on atmospheric pollutant concentrations in an industrial zone, namely Falconara, located on the central Adriatic coast of Italy. The aim of our study was to evaluate the possibility of assessing the diurnal evolution of air pollutants by introducing the Representative Day technique in order to obtain information on chemical reactions and air pollution in poorly monitored areas. The use of the Representative Day allows the compilation of large amounts of data in a compact format that will supply meaningful information on the whole data set.     

Design and Synthesis of Pro-Apoptotic Compounds Inspired by Diatom Oxylipins

Oxylipins are a large and diverse family of fatty acid derivatives exhibiting different levels of oxidation of the carbon chain. They are involved in many biological functions in mammals, plants and diatoms. In this last group of organisms, they are suggested to play a role in the reproductive failure of copepod predators, showing clear pro-apoptotic effects on newborn nauplii. In this work, these compounds were tested for the ability to induce mitotic arrest in sea urchin embryos. We show for the first time that oxylipins have an increased efficacy in their corresponding methylated form. Natural oxylipins were also used as an inspiration for the rational design and synthesis of stable chemical analogs with apoptotic activity against tumor cell lines. This approach led to the synthesis of the linear C15-ketol (22) that was shown to induce apoptosis in human leukemia U-937 cells. These results are proof of the concept of the use of eco-physiological considerations as a platform to guide the search for novel drug candidates.     

Cage culture of the Pacific white shrimp Litopenaeus vannamei (Boone, 1931) at different stocking densities in a shallow eutrophic lake

Postlarvae of Litopenaeus vannamei were acclimated and stocked in lake-based cages at the following stocking densities: 10, 20, 30 and 40 shrimp m⁻². Another set of shrimp was stocked in concrete tanks as reference samples at 30 shrimp m⁻². Significant differences were observed among stocking densities throughout the 95-day culture. The final weight at harvest decreased with increasing stocking density: mean weights of 23.3, 15.8, 13.0, 10.9 and 14.6 g for the 10, 20, 30, 40 shrimp m⁻² and reference tanks were observed respectively. There were no significant differences in survival throughout the culture period, ranging between 69% and 77%. Daily growth rates (range: 0.11–0.24 g day⁻¹) and specific growth rates (range: 3.54–4.34%) also differed significantly among stocking densities, both increasing with decreasing stocking density. The feed conversion ratio in the cages did not differ among the stocking densities, ranging from 1.53 to 1.65. The relationship between stocking density and mean individual weight at harvest followed the equation y =81.06 x ^−0.54 ( R ²=0.938) and that of stocking density and production (in g m⁻²) is y =58.01 x ^−0.46 ( R ²=0.834).