Biosynthesis of 14C-phytoene from tomato cell suspension cultures (Lycopersicon esculentum) for utilization in prostate cancer cell culture studies

This work describes the development and utilization of a plant cell culture production approach to biosynthesize and radiolabel phytoene and phytofluene for prostate cancer cell culture studies. The herbicide norflurazon was added to established cell suspension cultures of tomato (Lycopersicon esculentum cv. VFNT cherry), to induce the biosynthesis and accumulation of the lycopene precursors, phytoene and phytofluene, in their natural isomeric forms (15-cis-phytoene and two cis-phytofluene isomers). Norflurazon concentrations, solvent carrier type and concentration, and duration of culture exposure to norflurazon were screened to optimize phytoene and phytofluene synthesis. Maximum yields of both phytoene and phytofluene were achieved after 7 days of treatment with 0.03 mg norflurazon/40 mL fresh medium, provided in 0.07% solvent carrier. Introduction of 14C-sucrose to the tomato cell culture medium enabled the production of 14C-labeled phytoene for subsequent prostate tumor cell uptake studies. In DU 145 prostate tumor cells, it was determined that 15-cis-phytoene and an oxidized product of phytoene were taken up and partially metabolized by the cells. The ability to biosynthesize, radiolabel, and isolate these carotenoids from tomato cell cultures is a novel, valuable methodology for further in vitro and in vivo investigations into the roles of phytoene and phytofluene in cancer chemoprevention.     

The future of fish passage science,engineering, and practice

Much effort has been devoted to developing, constructing and refining fish passage facilities to enable target species to pass barriers on fluvial systems, and yet, fishway science, engineering and practice remain imperfect. In this review, 17 experts from different fish passage research fields (i.e., biology, ecology, physiology, ecohydraulics, engineering) and from different continents (i.e., North and South America, Europe, Africa, Australia) identified knowledge gaps and provided a roadmap for research priorities and technical developments. Once dominated by an engineering‐focused approach, fishway science today involves a wide range of disciplines from fish behaviour to socioeconomics to complex modelling of passage prioritization options in river networks. River barrier impacts on fish migration and dispersal are currently better understood than historically, but basic ecological knowledge underpinning the need for effective fish passage in many regions of the world, including in biodiversity hotspots (e.g., equatorial Africa, South‐East Asia), remains largely unknown. Designing efficient fishways, with minimal passage delay and post‐passage impacts, requires adaptive management and continued innovation. While the use of fishways in river restoration demands a transition towards fish passage at the community scale, advances in selective fishways are also needed to manage invasive fish colonization. Because of the erroneous view in some literature and communities of practice that fish passage is largely a proven technology, improved international collaboration, information sharing, method standardization and multidisciplinary training are needed. Further development of regional expertise is needed in South America, Asia and Africa where hydropower dams are currently being planned and constructed.     

Effects of open- and closed-system temperature changes on blood O2-binding characteristics of Atlantic bluefin tuna (Thunnus thynnus)

We investigated the effects of open- and closed-system temperature changes on the O₂ affinity of Atlantic bluefin tuna (Thunnus thynnus) blood using in vitro methods essentially identical to those previously employed on tropical tuna species. Bluefin tuna blood has a general O₂ affinity (P ₅₀ = 2.6–3.1 kPa or 19–23 mm Hg at 0.5% CO₂) similar to that of skipjack tuna, yellowfin tuna, and kawakawa blood (P ₅₀ = 2.8–3.1 kPa at 0.5% CO₂) but significantly above that of bigeye tuna blood (P ₅₀ = 1.6–2.0 kPa at 0.5% CO₂). We therefore hypothesize that bluefin tuna are less tolerant of hypoxia than bigeye tuna. Further, we found the P ₅₀ of bluefin tuna blood to be slightly reduced by a 10°C open-system temperature increase (e.g., from 4.83 kPa at 15°C to 3.95 kPa at 25°C) and to be completely unaffected by a 10°C closed-system temperature change. Bluefin tuna blood, therefore, had a significantly reduced Bohr effect when subjected to the inevitable changes in P CO ₂ and plasma pH that accompany closed-system temperature shifts (0.04–0.09 Δlog P₅₀ΔpH⁻¹) compared with the effects of changes in plasma pH accomplished by changing P CO ₂ alone (0.81–0.94 Δlog P₅₀ Δ pH⁻¹). This response is similar to that of skipjack tuna blood, but different from yellowfin or bigeye tuna blood. During closed-system temperature changes at oxygen levels above P ₅₀, however, bluefin tuna blood showed a reversed temperature effect (i.e., P O ₂ decreased in response to an increase in temperature). Unlike in other tuna species, temperature effects on O₂ affinity of bluefin tuna whole blood were similar to those previously reported for hemoglobin solutions, suggesting that red cell-mediated ligand changes are not involved.     

Amphibian oncology.

Spontaneous neoplasia is rare in all three orders of Amphibia.Tumors are documented in most major organ systems, and some have various underlying etiologies, including viral infection, environmental contaminants, and genetic predisposition. Currently,treatment options are limited to removal of the predisposing condition(s), palliative care, surgical excision, and, when necessary,humane euthanasia may be elected. Neoplasia must be distinguished from common infectious, nonneoplastic conditions that can negatively impact population health. This article is a review of the more common types of neoplasia in amphibians, and includes clinically relevant information, such as biologic behavior,anatomy, associated etiologies, major differential diagnoses, and clinical management.     

Prophylactic use of decoquinate for infections with Cryptosporidium parvum in experimentally challenged neonatal calves

OBJECTIVE: To evaluate the effect of daily oral administration of decoquinate to neonatal calves experimentally challenged with various numbers of Cryptosporidium parvum oocysts. DESIGN: Clinical trial. ANIMALS: 75 calves. PROCEDURE: Calves were purchased from a commercial dairy during a 5-week period. Calves were housed in individual hutches and fed milk replacer with or without decoquinate (2 mg/kg [0.9 mg/lb per day]). Calves were randomly assigned to treatment and 1 of 5 challenge groups (0, 50, 100, 1000, or 10,000 C. parvum oocysts in 60 mL of saline [0.9% NaCl] solution administered p.o. on the day after arrival). Calves were maintained in the study for as long as 28 days. Calves were clinically assessed for diarrhea and dehydration. Fecal samples were submitted for oocyst enumeration 3 times each week. RESULTS: Treatment did not affect number of days to first watery feces (diarrhea), number of days to first oocyst shedding, or duration of diarrhea or oocyst shedding. Duration of oocyst shedding was significantly associated with challenge dose of oocysts administered to calves and number of days to first oocyst shedding. Duration of diarrhea and number of days to first oocyst shedding were significantly associated with week of arrival and number of days to first watery diarrhea. CONCLUSIONS AND CLINICAL RELEVANCE: Daily treatment with decoquinate at the dosage used in this study did not affect oocyst shedding or clinical signs associated with cryptosporidiosis. However, there was an indication that if the number of oocysts calves received could be reduced, then the duration of oocyst shedding and, hence, environmental loading of C. parvum oocysts could be reduced.     

Failure to protect goats following vaccination with soluble proteins of Sarcoptes scabiei: evidence for a role for IgE antibody in protection

Developing an anti-scabies vaccine is thought to be a feasible alternative to chemical control, since animals which have recovered from sarcoptic mange become resistant against mite reinfestation. The purpose of this study was to evaluate the protective value of immune responses developed in animals after immunisation with soluble mite proteins. Soluble proteins from Sarcoptes scabiei were extracted then subjected to ion exchange chromatography, and proteins from the column were eluted step-wise with 0%, 10%, 25% and 50% of 1 M solution of NaCl in a Tris buffer. Each protein fraction was concentrated and dialysed against PBS. To evaluate the immunogenicity of the fractions, 36 goats were allocated into six groups, group1 goats were unvaccinated, group 2 were vaccinated with intact soluble mite proteins, and groups 3-6 were vaccinated respectively with the fractionated proteins. Vaccinations were conducted four times with 1 mg protein/dose and 4-week intervals between vaccinations. One week after the last vaccination, all goats were challenged with approximately 2000 live mites on the auricles and infestations were allowed to progress for 6 weeks. The severity of lesions caused by the infestation was assessed throughout the study. The challenge caused mange or encrustation dermatitis in all animals and no differences in severity of lesions were observed between vaccinated and unvaccinated control goats. Vaccination with each fraction of the mite proteins invoked high levels of scabies-specific IgG in the serum of all animals but failed to induce specific IgE as determined by Elisa. In contrast, goats challenged experimentally with a primary or repeated mite challenge developed strong serum IgE and IgG antibody responses to Sarcoptes antigens. The latter animals were shown in a previous study to be resistant to reinfestation. The lack of immune protection in the vaccinated animals may be attributed to the absence of protective levels of IgE antibody, and the present findings indicate that allergens and IgE antibody is important in immunity to S. scabiei infection.     

Metal Concentrations in Organs of the Clam Amiantis umbonella and Their Use in Monitoring Metal Contamination of Coastal Sediments

The aim of this study was to evaluate the use of metal concentrations in clam organs to monitor metal contamination in coastal sediments. The concentrations of Cd, Cr, Cu, Hg, Ni, Pb, V, and Zn were measured in the kidneys, gonads, mantles, gills, digestive gland, and hearts of the infaunal clam Amiantis umbonella collected from a contaminated site near desalination and power plant discharges, and a reference site in Kuwait Bay. Metal concentrations in sediment and sediment pore water were also measured at the collection sites of individual clams at the contaminated site. The concentrations of all metals in all organs (except Zn in the digestive gland) were significantly higher in clams from the contaminated site than from the reference site. Metal concentrations in several organs in A. umbonella from the contaminated site were correlated with those in the sediments and pore waters to which they were exposed. However, fresh weights of gonads, gills, and mantles were significantly lower in clams from the contaminated site compared to the reference site, indicating that the observed elevated concentrations of metals in the organs of clams from the contaminated site largely reflect lower organ weights, rather than higher metal loads, and that these organs in A. umbonella and perhaps other clams are not appropriate for use as biomonitors of metal contamination. Metal concentrations in clam kidneys showed a wide dynamic range with respect to environmental contamination and kidney weight was not variable. Therefore, metal concentrations in clam kidneys provide a reliable biomonitor of contaminant metals in coastal marine sediments.     

Effect of varying density and water level on the spawning response of African catfish Clarias gariepinus: Implications for seed production

The development of a reliable methodology for spawning of African catfish, Clarias gariepinus without the use of hormone injections would greatly improve the prospects of aquaculture in Africa. Earlier work has shown that it is possible to produce C. gariepinus fingerlings by subjecting the broodfish to a physical stress of reduced water depth and/or increased temperature. The hypothesis that C. gariepinus could be induced to spawn through a combined physical stress of lowered water level and increased stocking density was tested in concrete tanks. Three water levels (25, 50 and 75 cm) and three stocking densities (2, 4 and 6 pairs of broodfish at a 1:1 sex ratio in each hapa) were tested. Water depth in the tanks and brood fish density in the hapas affected spawning success. The percentage of spawning females was significantly higher when broodfish were stocked at 2 and 4 pairs in each hapa at water levels of 25 cm or 50 cm. There was no significant difference in spawning response between the 25 and 50 cm depths while a significant difference was seen between the 75 cm and both 25 and 50 cm depths. The results indicate optimum levels and densities for enhancing spawning success in C. gariepinus.     

The cell and molecular basis of mechanical, cold, and inflammatory pain

Peripheral pain pathways are activated by a range of stimuli. We used diphtheria toxin to kill all mouse postmitotic sensory neurons expressing the sodium channel Nav1.8. Mice showed normal motor activity and low-threshold mechanical and acute noxious heat responses but did not respond to noxious mechanical pressure or cold. They also showed a loss of enhanced pain responses and spontaneous pain behavior upon treatment with inflammatory insults. In contrast, nerve injury led to heightened pain sensitivity to thermal and mechanical stimuli indistinguishable from that seen with normal littermates. Pain behavior correlates well with central input from sensory neurons measured electrophysiologically in vivo. These data demonstrate that Na(v)1.8-expressing neurons are essential for mechanical, cold, and inflammatory pain but not for neuropathic pain or heat sensing.     

The selenium status of dairy herds in Prince Edward Island

Bulk tank milk selenium (Se) concentration was compared with mean serum Se concentration in 15 herds and was found to be an accurate reflection of the herd Se status. The Se status of 109 Prince Edward Island (PEI) dairy herds was monitored for 1 year using bulk tank milk Se concentration. Fifty-nine percent of the herds surveyed were, at some point, found to be marginal or deficient in Se, putting them at risk of disease and suboptimal production. The periods of greatest risk of deficiency were fall and winter, at which time 5% and 4%, respectively, of herds sampled fell in the range considered truly deficient in Se. Herds in which Se supplementation was provided in the form of a commercial dairy concentrate were over 4 times more likely to be Se-adequate than herds not using this method, and adjusted average daily milk yield was 7.6% greater in herds determined to be Se-adequate when compared with Se-marginal herds. We conclude that many dairy producers in PEI are providing insufficient supplementary Se in the ration to meet the recommended Se intake for lactating cows.