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.     

Comparative adverse cardiac effects of pimobendan and benazepril monotherapy in dogs with mild degenerative mitral valve disease: a prospective, controlled, blinded, and randomized study

BACKGROUND: Pimobendan (PIMO) is an inodilator that may have some beneficial effects in canine degenerative mitral valve disease (MVD). However, little information is available about its cardiac effects in dogs without systolic myocardial dysfunction. HYPOTHESIS: Compared to benazepril (BNZ), an angiotensin-converting enzyme inhibitor, PIMO may worsen valve regurgitation in early canine MVD. ANIMALS: Twelve Beagles with asymptomatic MVD were randomized into 2 groups (n = 6) receiving BNZ or PIMO at dosages of 0.25 mg/kg PO q24h and q12h respectively, for 512 days. METHODS: The study followed a blinded, randomized, prospective, and parallel group design. After day 512, the dogs were necropsied, and cardiac histopathology was performed in a blinded manner. RESULTS: A significant treatment effect was observed as soon as day 15 with increased systolic function in the PIMO group by comparison to baseline value as assessed by fractional shortening (P < .0001) and tissue Doppler variables (P = .001). Concurrently, the maximum area and peak velocity of the regurgitant jet signal increased (P < .001), whereas these variables remained stable in the BNZ group. Histologic grades of mitral valve lesions were more severe in the PIMO group than in the BNZ group. Moreover, acute focal hemorrhages, endothelial papillary hyperplasia, and infiltration of chordae tendinae with glycosaminoglycans were observed in the mitral valves of dogs from the PIMO group but not in those of the BNZ group. CONCLUSIONS AND CLINICAL IMPORTANCE: PIMO has adverse cardiac functional and morphologic effects in dogs with asymptomatic MVD. Additional investigation in dogs with symptomatic MVD is now warranted.     

Quantitative assessment of velocities of the annulus of the left atrioventricular valve and left ventricular free wall in healthy cats by use of two-dimensional color tissue Doppler imaging

OBJECTIVE: To analyze velocities of the annulus of the left atrioventricular valve and left ventricular free wall (LVFW) in a large population of healthy cats by use of 2-dimensional color tissue Doppler imaging (TDI). ANIMALS: 100 healthy cats (0.3 to 12.0 years old; weighing 1.0 to 8.0 kg) of 6 breeds. PROCEDURE: Radial myocardial velocities were recorded in an endocardial and epicardial segment, and longitudinal velocities were recorded in 2 LVFW segments (basal and apical) and in the annulus of the left atrioventricular valve. RESULTS: LVFW velocities were significantly higher in the endocardial than epicardial layers and significantly higher in the basal than apical segments. For systole, early diastole, and late diastole, mean +/- SD radial myocardial velocity gradient (MVG), which was defined as the difference between endocardial and epicardial velocities, was 2.2 +/- 0.7, 3.3 +/- 1.3, and 1.8 +/- 0.7 cm/s, respectively, and longitudinal MVG, which was defined as the difference between basal and apical velocities, was 2.7 +/- 0.8, 3.1 +/- 1.4, and 2.1 +/- 0.9 cm/s, respectively. A breed effect was documented for several TDI variables; therefore, reference intervals for the TDI variables were determined for the 2 predominant breeds represented (Maine Coon and domestic shorthair cats). CONCLUSIONS AND CLINICAL RELEVANCE: LVFW velocities in healthy cats decrease from the endocardium to the epicardium and from the base to apex, thus defining radial and longitudinal MVG. These indices could complement conventional analysis of left ventricular function and contribute to the early accurate detection of cardiomyopathy in cats.     

Remediation Actions by a Risk Assessment Approach: A Case Study of Mercury Contamination

The risk assessment procedure for identifying the remediation actions which may be adopted at a mercury contaminated site, when the plants are upgraded in the future, is proposed. The potentially active exposure/migration pathways in the future arrangement of the area will be due to Hg contaminated subsoil as a primary source (vapor inhalation and groundwater leaching) and to groundwater as a possible secondary source (transport to the point of compliance). The data of mercury concentration in the soil were acquired through environmental monitoring campaigns, and were processed to establish the three-dimensional distribution of contamination in subsoil, to locate sources and to define their geometrical and chemical characteristics. Speciation tests of mercury in the soil indicated that the most abundant species present were poorly leachable under the site-specific environmental conditions, confirming the coefficient distribution value obtained by the leaching tests. Analytical and numerical fate and transport modeling tools were used to locate digging zones in the contaminated subsoil, so as to reduce the possible groundwater contaminant loading and to avoid the down-gradient exceeding the concentration limit according to regulations. Remediation actions additional to civil works were required, which consists of soil digging within one contamination source, for about 22,200 m³ of soil. In order to evaluate the Hazard Index (HI) for human receptors due to Hg vapor inhalation, the air concentration of volatile mercury at the exposure point was estimated, based on direct measurements carried out at the site. Simulation gave HI values below 1 for all tested scenarios, suggesting that public health is protected without any additional actions to the already scheduled plant upgrading and digging for groundwater protection.     

Intraoperative contrast echocardiography to verify the surgical occlusion of a single extrahepatic portosystemic shunt in a dog

This report details a bubble echocardiographic study carried out during the surgical treatment of a congenital single extrahepatic portosystemic shunt (PSS) in a Labrador Retriever. After celiotomy, agitated saline was injected through a jejunal vein and microbubbles appeared rapidly in the right cardiac chambers. The test confirmed the presence of a PSS, helping the surgeon to identify the vessel concerned and to rule out a second shunt. Successively, portography confirmed what the exploratory celiotomy had revealed before with the aid of the bubble study: a single shunt was located between the portal vein and the right renal vein. It was completely ligated, as all the criteria for this solution were met. Intraoperative contrast echocardiography (ICE) was easy to perform, helpful and undemanding. It is proposed here as an intraoperative ancillary test to diagnose all PSS and to confirm successful treatment when complete shunt closure is possible.     

Sequential solidification/stabilization and thermal process under vacuum for the treatment of mercury in sediments

Purpose  

Millions of cubic meters of sediments are dredged every year in the world. About 10–20% on weight basis of this material is contaminated by organic and/or inorganic pollutants. This work presents the laboratory tests performed to study a system for the remediation and reuse of mercury-contaminated sediments. The treatment is based on a cement-based granulation step (solidification/stabilization (S/S)), followed by a thermal process under vacuum during which volatile and semi-volatile compounds are removed. The experiments focused on: (1) cement hydration reactions; (2) pollutant removal efficiencies; and (3) leaching behavior, in relation to temperature and duration of the thermal process. Mercury speciation was also investigated.     

Polycyclic Aromatic Hydrocarbons (PAHs) Slurry Phase Bioremediation of a Manufacturing Gas Plant (MGP) Site Aged Soil

Bench scale tests have been carried out in order to investigatebioremediation feasibility of a Manufacturing Gas Plant site(Bovisa Gasometri – MI – I) aged soil, highly contaminated bypolyaromatic hydrocarbons (PAHs) and mineral oils. Biodegradationstudies were carried out at 22 °C in a slurry system reactor, with a solid to liquid ratio of 10% w/w. Three testswere performed, over a period of 23, 24 and 91 days respectively.In the first test, only soil and water were put into the system.In the second test, microbial activity was inhibited to evaluatethe amount of abiotic losses. In the third test, macronutrientswere added to the reactor; bioaugmentation was also carried outby an inoculum of autochthonous PAH-degrading microorganisms. Saturated hydrocarbons, PAHs, transformation products, heterotropic and PAH-degrading bacteria and fungi were monitored.Tests showed that high removal efficiency could be obtained following 23 days of treatment for all PAHs, including high molecular weight compounds. Abiotic losses were an importantprocess contributing to concentration abatement in soil, especially in the first phase of the treatment. Lag time lackfor all polyaromatic hydrocarbons revealed that autochthonousmicroorganisms were well adapted to these contaminants; bioaugmentation did not seem to speed up the process. The higherremoval rate and efficiency for high molecular weight PAHs obtained in the third test suggested that nutrient addition could play an important role in the biodegradation process ofthese compounds, whereas it did not significantly modify total(abiotic + biotic) removal of light compounds.     

Lab-scale feasibility tests for sediment treatment using different physico-chemical techniques

Purpose  

The fairly high amounts of sediments dredged in coastal or internal water bodies for navigational and/or environmental purposes claims for the identification of appropriate management strategies. Dredged sediments are frequently affected by organic and inorganic contamination, so that their reuse, as an alternative to final landfill disposal, could need remediation. In this framework, a two-year joint research project was carried out to assess the feasibility of different remediation technologies for the treatment of polluted sediments.