Auranofin improves overall survival when combined with standard of care in a pilot study involving dogs with osteosarcoma

Osteosarcoma is the most common paediatric primary bone malignancy. The major cause of death in osteosarcoma is drug‐resistant pulmonary metastasis. Previous studies have shown that thioredoxin reductase 2 is a driver of metastasis in osteosarcoma and can be inhibited by auranofin (AF). Moreover, studies have shown that AF significantly reduces pulmonary metastases in xenotransplant models. Here, we describe a phase I/II study of AF in canine osteosarcoma, a well‐recognized spontaneous model of human osteosarcoma. We performed a single‐arm multicentre pilot study of AF in combination with standard of care (SOC) (amputation + carboplatin). We recruited 40 dogs to the trial and used a historical SOC‐only control group (n = 26). Dogs >15 kg received 9 mg AF q3d PO and dogs <15 kg received 6 mg q3d. Follow‐up occurred over at least a 3‐year period. Auranofin plus SOC improved overall survival (OS) (P = .036) in all dogs treated. The improved outcome was attributable entirely to improved OS in male dogs (P = .009). At the time of writing, 10 dogs (25%) survive without measurable disease in the treatment group with survival times ranging between 806 and 1525 days. Our study shows that AF improves OS in male dogs when combined with SOC. Our findings have translational relevance for the management of canine and human osteosarcoma. Our data justify a larger multicentre phase 2 trial in dogs and a phase I/II trial in human patients with refractory disease at the time of initial surgery.     

Effect of diet composition on nutrient digestibility and digestive enzyme levels of gilthead sea bream (Sparus aurata L.)

An experiment was conducted to evaluate the effects of dietary composition on apparent digestibility coefficients of protein, fat and carbohydrate of gilthead sea bream. In addition α‐amylase and proteolytic activities were measured in the digestive tract of fish, held at 20±1°C. Six experimental diets were formulated containing approximately 40%, 45% and 50% protein, 11% and 21% fat and a starch level which fluctuated from 14% to 36%. Fish having an average weight of 100–130 g were used, fed at 1.5% of their body weight daily, and digestibility was measured using an indirect method. Enzyme activities were measured in the digestive tract of fish, fed diets containing a combination of 40%, 50% protein with 11%, 21% fat at 0.5, 5, 10, 24 h after a single meal. Starch digestibility was reduced with its level in the diet. It also negatively affected fat digestibility as well as protein digestibility, the last only at the high fat level. Dietary fat level had a negative effect on starch and protein digestibility. Fat affected also strongly α‐amylase levels in the digestive tract, while its effect on protease activity was of smaller magnitude. These results indicate that digestive enzyme activities and nutrient digestibility values are affected by dietary composition, carbohydrates and fat indicating the strongest effect.     

Stability of virgin olive oil. 1. Autoxidation studies

Virgin olive oil samples with similar oxidative stabilities and fatty acid compositions were stored for 24 months. Changes in the lipid substrate were followed by peroxide value and K(232) measurements. HPLC was used to evaluate changes in the alpha-tocopherol, pigment, and squalene contents. Total polar phenol content was measured colorimetrically. The loss of alpha-tocopherol and carotenoids was comparable with that of polar phenol content, suggesting an active participation in autoxidation. The limited role of squalene in autoxidation was further confirmed using an olive oil model and in the presence of alpha-tocopherol. Pheophytin a degradation was high, although spectrometric estimation of chlorophyll content did not indicate so. Evaluation of pheophytin a activity at three different levels of addition on the oil model indicated a concentration-dependent antioxidant role more pronounced at elevated temperatures, which could be partially due to the activity of certain degradation products.     

Stability of virgin olive oil. 2. Photo-oxidation studies

Virgin olive oil samples with similar oxidative stabilities and fatty acid compositions were exposed to 12100 lx (25 +/- 1 degrees C) in closed bottles until bleached. The observed low changes in the substrate and polar phenols were related to oxygen availability. HPLC monitoring showed that pheophytin agradual degradation (> 90%) was accompanied by a considerable alpha-tocopherol loss (22-35%) due to the reaction of the latter with singlet oxygen. No changes were recorded for carotenoids, which acted as physical quenchers and light filters. Squalene loss was confined (4-12%). Complementary experiments on the activity of pheophytin a, using olive oil models, indicated a concentration dependence, enhanced by oxygen availability. In closed bottles, the degradation rate constant was higher at low amounts of pheophytin a. Squalene was preferentially consumed to protect alpha-tocopherol. An urgent change in the practice of packaging is needed to preserve the precious characteristics of the product during commercialization.     

Rapid, fully automated flow injection antioxidant capacity assay

A flow injection method for antioxidant capacity assessment based on a low-cost laboratory-made analyzer is reported. A sample of 30 microL is injected in acetate buffer stream, pH 4.6, that converges with ABTS*(+) reagent stream. Detection is achieved by monitoring absorbance at 414 nm. The proposed method achieves a sample throughput of up to 120 samples h(-1), the detection limit being 1.3 microM trolox. Precision was better than 5% relative standard deviation (n = 4) and the linear range was 4-100 microM, expanded to 250 microM trolox utilizing concentration gradients formed along the injected sample bolus. Information on reaction kinetics is obtained through a single injection. The method was applied to pure compounds and wine and honey samples. Good correlation was found between antioxidant capacity assessed through the proposed method and phenolic content: r = 0.94 for red wines, r = 0.96 for white and rose wines, and r = 0.89 for honeys.     

The Impact of Cl ?? Cd Interrelationship on Planning Wastewater Reuse in Cabbage

Two greenhouse pot experiments were conducted in Agrinion, Greece, using a randomized block design in four replications, respectively, as follows: The first one included five levels of treated municipal wastewater (TMWW), being used as an irrigation water source. The second one, five levels of applied Cl at a constant soil applied Cd level of 10.36?mg/kg soil, the plants being irrigated with fresh well water. The purpose of these experiments was to study the impact of the Cl ?? Cd interrelationship on planning TMWW reuse, for the irrigation of Brassica oleracea var. Capitata (cabbage) cv F1 Gloria, ehich was used as test plant, in both of these experiments. It was found that the TMWW Cl content, originating mainly from the procedure of wastewater chlorination, was synergistically interrelated with the toxic heavy metal Cd, increasing its soil availability and cabbage plant leaf uptake (edible plant part). As this increase is directly associated with the consumer??s health, it was suggested that the TMWW be subjected to dechlorination process or the disinfection be made by ozonation or UV, which do not include Cl.     

Yolk‐sac larval development of the substrate‐brooding cichlid Archocentrus nigrofasciatus in relation to temperature

In order to conserve and culture the cichlid fish Archocentrus nigrofasciatus, more information about its reproductive biology and its larval behavior and morphogenesis is necessary. Currently, temperatures ranging from 21 to 27 °C are used in ornamental aquaculture hatcheries. Lower temperatures are preferred to reduce the costs of water heating, and 23 °C is usually the selected temperature. However, there is limited information on culturing protocols for ornamental species and most of the information generated on this topic remains scarce. Thus, the present study examines the morphological development of Archocentrus nigrofasciatus during the yolk‐sac period up to the age of 100 h post‐hatching in relation to 2 temperature regimes used in ornamental aquaculture: a temperature of 27 °C (thermal optimum) and a decreased temperature of 23 °C (thermal tolerance). The results of this study suggest that the 27 °C temperature generates intense morphological changes in yolk‐sac development in a shorter period. This has advantages as it reduces the time of yolk‐sac larval development, and, thus, minimizes the transition phase to exogenous feeding and maximizes the efficiency at which yolk is converted into body tissues. The present paper provides necessary information to produce freshwater ornamental fish with better practices so as to increase larval survival and capitalize on time for growth.     

Recovery of squalene from wine lees using ultrasound assisted extraction-a feasibility study

The present work is a systematic approach for valorization of wine lees regarding the recovery of squalene, a bioactive lipid. Such a study is presented for the first time in literature. Separate examination of squalene content in "light" and "heavy" lees from different vinification processes by RP-HPLC demonstrated that these waste streams can be used as a source for this lipid, despite variations due to technological or genetic effects. Next, ultrasound assisted extraction of squalene from the "industrial waste" (the mixture of wine lees generated from different wines) using n-hexane was optimized with the aid of response surface methodology (independent variables: sonication duration and duty cycles). Autolysis was monitored through optical microscopy. Squalene yield (0.6 ± 0.08 g SQ/kg dry lees) was comparable to that of recently examined potential sources (0.2-0.35 g SQ/kg dry olive pomace and 0.06 g SQ/kg olive leaves).     

Nutritional quality of greenhouse lettuce at harvest and after storage in relation to N application and cultivation season

The effect of five levels of nitrogen fertilization on the growth and nutritional quality of Cos lettuce (Lactuca sativa L. cv. Parris Island) at harvest and after storage was studied during autumn and winter in South-West Greece. Plants were cultivated hydroponically in a greenhouse and the nitrate, chlorophyll and ascorbic acid (vitamin C) concentrations within the plant tissues were measured at harvest and following storage at 5 or 10 °C for 10 days. Nitrate accumulated in the leaves with increasing amounts of N within the nutrient solution and was higher in the winter than in the autumn. At the lowest N level (20 mg L⁻¹), the inner leaves accumulated more nitrate than the outer leaves, whereas at higher N levels (140, 200 or 260 mg L⁻¹) nitrate accumulation was higher in the outer leaves. Overall, the highest nitrate concentrations were detected in the petiole and the proximal end of the leaf, but at the lowest N application rate (20 mg L⁻¹) nitrate accumulated in the distal region of the leaf too. Although the nitrate concentrations within the leaves did not change significantly during 10 days storage at 5 or 10 °C, the chlorophyll and vitamin C concentrations decreased. Chlorophyll loss was higher in lettuce that was grown under low N levels and was higher at 10 °C than at 5 °C, but was reduced by enclosure of the lettuce in polyethylene film. It is concluded that the optimum N application rate for Cos lettuce grown hydroponically under cover during autumn and winter in South-West Greece, and in other areas with a similar climate, is 200 mg N L⁻¹ because at this N rate yield is satisfactory and leaf nitrate concentrations are below the maximum acceptable level for human consumption. Nutritional value (vitamin C concentration) and market quality (chlorophyll content) are highest at harvest and decrease during storage, but quality in terms of nitrate concentration does not change.     

Evaluation of zinc supplementation in European sea bass (Dicentrarchus labrax) juvenile diets

This study aimed to investigate the effect of increased zinc dietary levels on the growth performance, feed utilization, immune status and induced wound healing in European sea bass (Dicentrarchus labrax). Fish weighing 10 g were fed five diets containing organic zinc at 30, 70, 110 and 150 mg kg^?1 diet and one inorganic zinc source (zinc oxide) at 150 mg kg^?1 diet for a period of 12 weeks. No significant (P>0.05) differences were found in the growth performance parameters (final weight, specific growth rate), immunological indices tested (respiratory burst activity in whole blood) and the wound‐healing process. Zinc accumulation in the skin, vertebrae and liver increased significantly (P<0.01) with increased dietary zinc levels but not in muscle. The highest zinc concentrations were obtained in skin tissue, followed by vertebrae, liver and muscle tissue. Using the concentration of zinc in skin as a response criterion, broken‐line analysis showed that the supplementation of 148 mg organic Zn kg^?1 diet seemed to be the optimum dietary zinc supplementation level for sea bass juveniles.