Cytotoxic Effects of Loperamide Hydrochloride on Canine Cancer Cells

Loperamide is a peripheral opiate agonist that can cause apoptosis and G2/M arrest in human cancer cell lines and may sensitize cells to chemotherapy. The objectives of this study were to investigate the effects of loperamide on viability, apoptosis and cell cycle kinetics in canine cancer cells and to establish whether the drug sensitizes cells to doxorubicin. Cell viability was assessed using Alamar Blue. Cell death and cell cycle were studied using flow cytometry with 7-Aminoactinomycin-D (7-AAD) and propidium iodide (PI), respectively. Loperamide decreased cell viability in a dose-dependent fashion and was most effective against canine osteosarcoma cells. In all cell lines, it induced a dose and time dependent apoptosis and resulted in accumulation in G0/G1. When co-incubated with doxorubicin, loperamide induced a synergistic cell kill in canine carcinoma cells. Investigation is warranted into the role of loperamide in the treatment of canine cancer.     

Relationship of Progesterone,Bovine Pregnancy-Associated Glycoprotein-1 and Nitric Oxide with Late Embryonic and Early Fetal Mortalities in Dairy Cows

The aim of the present study was to determine the relationship of progesterone (P4), bovine pregnancy-associated glycoprotein-1 (bPAG-1) and nitric oxide (NO) levels with late embryonic (LEM; day 28 to day 42) and early fetal mortalities (EFM; > day 42 to day 56) in dairy cows. Transrectal ultrasonography (6–8 MHz) was performed in 100 Holstein-Friesian cows at days 28, 42 and 56 after artificial insemination (AI; day 0) to diagnose pregnancy and to monitor the fate of the embryo. After ultrasound scanning of each cow, a milk sample was collected for assessment of P4 by an ELISA test and a blood sample was collected for assessment of bPAG-1, by using a double-antibody radioimmunoassay, and serum NO metabolites (nitrate + nitrite). Based on ultrasonographic examinations and bPAG-1-RIA, 41 of 100 inseminated cows were confirmed pregnant at day 28 after AI. Nine cows suffered of LEM, and 6 cows suffered of EFM and the overall pregnancy loss rate was 36.6% (15/41) between days 28 and 56 of pregnancy. By logistic regression analysis, there were no significant relationships between the level of P4 and bPAG-1 at day 28 after AI and the occurrence of LEM and EFM. Also, there were no significant relationships between the levels of P4 and bPAG-1 at day 42 and the occurrence of EFM. On the other hand, a significant relationship (P<0.05) was found between NO level at day 28 and the occurrence of LEM. In conclusion, measurement of the serum NO concentration at day 28 of pregnancy might help to predict the outcome of pregnancy by day 42 in dairy cows but further studies are needed to confirm this.     

MALDI‐MS Lipid Profiles of Oocytes Recovered by Ovum Pickup from Bos indicus and 1/2 indicus × taurus with High vs Low Oocyte Yields

The aim of the present study was to compare the lipid profile in oocytes of indicus and 1/2 indicus × taurus cows with high and low antral follicle count (AFC)/oocyte yields. After an OPU procedure (D0), antral follicles ≥3 mm were counted by ultrasonography (D4, 19, 34, 49, 64), and cows were assigned to groups with either high AFC (≥30 follicles; indicus, NH group; 1/2 indicus × taurus, AH group) or low AFC (≤15 antral follicles; indicus, NL group; 1/2 indicus × taurus, AL group). The lipid profiles of the oocytes were determined by MALDI‐MS. For GI, GII and GIII oocytes, the indicus samples tend to cluster separately from the 1/2 indicus × taurus samples. The lipid species [PC (P‐38:5) + H]⁺ and/or [PC (P‐36:2) + Na]⁺, [PC (38:2) + H]⁺, [PC (38:5) + Na]⁺ and [TAG (60:8) + NH⁴]⁺ were more abundant in indicus (NH and NL groups) than 1/2 indicus × taurus. The higher lipid content in the indicus oocytes likely reflects differences in the rate of lipid metabolism and may contribute to oocyte competence and embryo development.     

Content of plasmatic glycerol and activity of hepatic glycerol kinase in broiler chickens fed diets containing different sources and concentrations of glycerine

The objective of this study was to evaluate the effect of three sources of glycerine (crude glycerine from soya bean oil – CGSO, mixed crude glycerine from frying oil and lard – MCG, and a semipurified glycerine from soya bean oil – SPGSO) in four concentrations in the diet (17.5, 35.0, 52.5 and 70.0 g of each type of glycerine per kg of feed) on the levels of plasmatic glycerol and the activity of the hepatic enzyme glycerol kinase in broilers of 22–35 days old (experiment I) and 33–42 days old (experiment II). The highest (p < 0.05) plasmatic glycerol level was detected in broilers fed diet containing CGSO. Independent of the source, increasing the concentration of glycerine led to a linear increase (p < 0.05) in the plasmatic glycerol concentration. In experiment I, all the diets containing glycerine resulted in increased concentrations of plasmatic glycerol, in relation to the control diet without glycerine. However, in experiment II, only the diet containing 17.5 g of CGSO per kg of feed and the diets formulated with any of the three types of glycerine in the inclusion concentrations of 35, 52.5 and 70 g/kg of feed resulted in higher plasmatic glycerol levels than those observed in the control broilers. The source of glycerine influenced the glycerol kinase activity only in experiment II, where the use of CGSO in the diet increased the enzyme activity. For both experiments, the glycerol kinase activity increased with the inclusion of glycerine in the diet. In conclusion, based on the absence of saturation of the glycerol kinase activity for the three glycerine sources and for both rearing periods evaluated, the broilers can metabolize the glycerol (at the level of the phosphorylation of the glycerol to glycerol‐3‐phosphate in the liver) present in the glycerine when the diet is supplemented with up to 70 g of glycerine per kg of feed.     

US strategy for forest management adaptation to climate change: building a framework for decision making

Context

Recent policy changes in the USA direct agencies managing federal forests to analyze the potential effects of climate change on forest productivity, water resource protection, wildlife habitat, biodiversity, and other values.

Aims

This paper describes methods developed to (1) assess current risks, vulnerabilities, and gaps in knowledge; (2) engage internal agency resources and external partners in the development of options and solutions; and (3) manage forest resources for resilience, not just in terms of natural ecosystems but in affected human communities as well.

Methods

We describe an approach designed to characterize certain climate change effects on forests, and estimate the effectiveness of response options ranging from resistance to a realignment of management objectives.

Results

Field testing on a 6,300 km² area of conifer forest in the northwestern USA shows this decision model to be useful and cost-effective in identifying the highest sensitivities relating to vegetation management, biological diversity, water resources and forest transportation systems, and building consensus for adaptive strategies and actions.

Conclusions

Results suggest that this approach is an effective means for guiding management decisions to adapt to the effects of climate change, and provides an empirical basis for setting budgetary and management priorities.     

Nitrogen fertilization of black walnut (Juglans nigra L.) during plantation establishment. Physiology of production

Physiological mechanisms by which nitrogen (N) fertilization affects growth and development in temperate deciduous forest trees are not clearly understood, especially under intensive silvicultural systems. Grafted, Tippecanoe 1 cultivar black walnut (Juglans nigra L.) trees were grown in an intensively managed plantation in west-central Spain and subjected to six, fixed-nutrient-ratio complete fertilizer treatments (defined as 0, 25, 50, 75, 150, and 300 g N tree^?1) delivered via daily fertigation. Leaf chemistry and morphology were evaluated from June to September, and gas exchange was measured in July. Specific leaf mass, leaflet nitrogen (N), and chlorophyll concentrations varied over the course of the growing season, yet consistently increased with increasing fertilization. Net photosynthesis at ambient (A _net) and light-saturated (A _max) conditions increased from the unfertilized control to lowest treatment (25 g N) but did not increase at higher fertilizer rates. Photosynthetic N and chlorophyll use efficiencies decreased with increasing fertilization, but photosynthetic phosphorus and water use efficiencies increased. Transpiration rates and dark respiration were not significantly affected by treatment. Overall, the lowest fertilizer treatment (25 g N) had the greatest photosynthetic efficiency. Interactions between N and other nutrients with increasing fertilizer application suggested potential for nutrient imbalances at high fertilization rates. Our results provide a physiological justification for the use of low-to-moderate fertilization as an efficient strategy to promote black walnut plantation establishment under intensive cultural systems.     

Benthic microbial indicators of fish farm impact in a coastal area of the Tyrrhenian Sea

We studied the impact of organic loads due to the biodeposition of a fish farm in a non-impacted coastal area of the Tyrrhenian Sea (Western Mediterranean). Sediment chemistry and benthic microbial community were investigated from July 1997 to February 1998 on monthly basis at two stations: one was located under the fish farm, while the second was about 1 km away, and served as a reference site. The presence of the cage induced rapid changes in the benthic conditions: the sediments were rapidly (after 6 weeks) reduced. A significant accumulation of biopolymeric carbon was observed beneath the cage both 2 weeks after the initial cage deployment, on non-impacted sediments, and 5–7 months after, and appeared to be related to the fish farm production cycle. The density of microbial communities beneath the fish farm increased only during the first month of farming activity (July) and partially 7 months after cage disposal (i.e. in January), when there was a significant sediment organic enrichment. Additional evidence of rapid impact of the fish farm on the benthic bacterial communities is provided by the increase in the numbers of autofluorescent microbial cells. The photosynthetic eukaryotic cells displayed a highly reduced contribution to total autofluorescent microbial density, indicating that their decrease is related with biopolymeric carbon accumulation, and probably was due also to “shadow effect” induced by suspended material, coming from the fish farm, on the environment below. We propose here, to apply the ratio of culturable heterotrophic bacteria to microbial direct counts (CFU/MDC) to detect fish farm impact. In cage sediments, organic enrichment and the consequent modification of the characteristics of the benthic environment, determined an increase in aerobic heterotrophic bacteria and vibrio density indicating that they are efficient colonizers of organic-rich sediments. Densities of Escherichia coli and Enterococci were not significantly higher than in the reference site, and are likely of terrestrial origin. Thus they cannot be used as specific indicators of fish farm impact.     

Feasibility and repeatability of cold and mechanical quantitative sensory testing in normal dogs

Feasibility and inter-session repeatability of cold and mechanical quantitative sensory testing (QST) were assessed in 24 normal dogs. Cold thermal latencies were evaluated using a thermal probe (0 °C) applied to three pelvic limb sites. Mechanical thresholds were measured using an electronic von Frey anesthesiometer (EVF) and a blunt-probed pressure algometer (PA) applied to the dorsal aspect of the metatarsus. All QST trials were performed with dogs in lateral recumbency. Collection of cold QST data was easy (feasible) in 19/24 (79%) dogs. However, only 18.4%, 18.9% and 13.2% of cold QST trials elicited a response at the medial tibia, third digital pad and plantar metatarsal regions, respectively. Collection of mechanical QST data was easy (feasible) in 20/24 (83%) dogs for both EVF and PA.At consecutive sampling times, approximately 2 weeks apart, the average EVF sensory thresholds were 414 ± 186 g and 379 ± 166 g, respectively, and the average PA sensory thresholds were 1089 ± 414 g and 1028 ± 331 g, respectively. There was no significant difference in inter-session or inter-limb threshold values for either mechanical QST device. The cold QST protocol in this study was achievable, but did not provide consistently quantifiable results. Both mechanical QST devices tested provided repeatable, reliable sensory threshold measurements in normal, client-owned dogs. These findings contribute to the validation of the EVF and PA as tools to obtain repeated QST data over time in dogs to assess somatosensory processing changes.     

Oyster greening by outdoor mass culture of the diatom Haslea ostrearia Simonsen in enriched seawater

Oyster greening was first described in the seventeenth century as a natural phenomenon. However, it has recently been discovered that the diatom Haslea ostrearia Simonsen causes greening by synthesis of a blue pigment designated as ‘marennine’. This phenomenon, which involves massive proliferation of H. ostrearia in oyster ponds, was not understood or controlled by oyster farmers in the Marennes‐Oléron region (Atlantic coast of France). As greening oysters improved their market value, they tried to develop empirical methods to guarantee oyster fattening and improve profits. In this context, the present study investigated the feasibility of mass culture of diatoms outdoors in 10‐m³ ponds, using enriched seawater. Different biotic and abiotic parameters were monitored daily to determine the influence of the day–night temperature range. After 8 days, H. ostrearia was the dominant diatom species (66%), reaching a mean cell concentration of 2 × 10⁵ cell mL^?1 and a marennine concentration of 3.4 mg L^?1. Although intensive greening was obtained, further studies are required to optimize the production stages before this technology can be transferred to oyster farmers.