Culture of Differentiated Adult Rabbit Auricular Chondrocytes

Chondrocytes dedifferentiate to a fibroblast‐like phenotype on plastic surfaces. Dedifferentiation is reversible if these cells are then cultured embedded in gels as alginate, agarose or collagen. Chondrocytes cultured in suspension on a non‐adherent surface are also known to form aggregates of differentiated cells. The knowledge of chondrocyte behavior in culture is relevant for tissue engineering purposes. In this report we describe a simple method to culture differentiated or redifferentiated rabbit auricular chondrocytes on plastic surfaces with a stable phenotype. When chondrocyte aggregates formed in suspension are next seeded on plastic surfaces, most of them attach to the plastic as round or polygonal cells, and this morphological differentiation, confirmed by the presence of type II collagen, is stable for long culture periods. We also report that the addition of aggregates to monolayer cultures of dedifferentiated chondrocytes results in their redifferentiation, as is shown by their morphological changes and the synthesis of type II collagen. Therefore, this simple method can be useful for the study of chondrocyte behavior on plastic surfaces and for redifferentiating previously proliferated chondrocytes in tissue engineering techniques. Furthermore, these results demonstrate that, in addition to culture conditions such as cell isolation method or cell‐density, chondrocyte behavior on plastic depends on the presence or absence of aggregates resulting from the dissociation process.     

Occurrence and Distribution of Lactate Dehydrogenase in Knee Joint Carilage in Healthy and Osteoarthrotic Dogs

According to clinical studies, degenerative diseases of canine joints lead to higher lactate dehydrogenase (LDH) levels in synovial fluid. The goal of the present study was to examine the intraarticular distribution of LDH in healthy and osteoarthrotic knee joints in order to identify possible sources of LDH in synovial fluid. As synovial LDH concentrations neither correlate with the number of leukocytes nor with synovitis, our investigation focused on the articular cartilage. Samples from healthy and osteoarthrotic knee joints were fixed and processed for transmission electron microscopy (TEM), immunohistochemistry (IHC), and immunocytochemistry (ICC). In addition, fresh cartilage samples were investigated cytochemically by the tetrazolium‐formazan reaction. Analyses of blood and synovial fluid samples were used to confirm the absence of inflammatory disease. Morphology of articular cartilage was assessed macroscopically and by means of TEM. IHC revealed highest levels of LDH in chondrones and a diffuse labelling of the matrix with a distinctive decrease in signal from superficial to deeper cartilage layers. Ultrastructural localization by ICC showed LDH to be present in the cytoplasm of all chondrocytes and confirmed the density gradient in the matrix. Labelling was absent from nuclei and from pericellular rims. Cytochemistry confirmed the distribution pattern and, thus, expanded our findings beyond immunological evidence by providing proof of enzymatic activity of LDH in articular cartilage. The present results indicate that LDH is transferred from chondrocytes to the cartilaginous matrix. We suggest, therefore, that LDH found in synovial fluid originates from the articular cartilage and that osteoarthrotic processes promote LDH release from the cartilaginous matrix.     

Distribution and Activity of Fatty Acid Amide Hydralase (FAAH) in the Gastrointestinal Neurons of Mouse

The endocannabinoid anandamide may regulate intestinal motility through activation of CB1 receptors. Anandamide is then inactivated by fatty acid amide hydrolase (FAAH), a membrane bound enzyme. Under pathological conditions, inactivation of such enzymatic activity may lead to inhibition of the intestinal motility. Here, preliminary reports on the distribution of Fatty Acid Amide Hydrolase (FAAH) immunoreactivity in the mouse gastrointestinal neurons, and the pharmacological effects of N‐arachidonoylserotonin (AA‐5HT), a selective inhibitor of FAAH, are reported. FAAH was revealed by an indirect immunofluorescence. Laminar preparations containing the myenteric or the submucous plexus adhered, were peeled off after the whole gut wall had been stretched out and fixed in 4% paraformaldehyde. They were subsequently incubated with a polyclonal anti‐serum directed against a region near the N‐terminus of the human FAAH and revealed by a FITC‐conjugated goat anti‐rabbit secondary anti‐serum. FAAH‐immunoreactive neurons were observed within the myenteric ganglia throughout the GIT. The positive nerve cells varied in size and density of immunoreactivity. Stomach and large intestine showed the highest neuronal density. AA‐5HT significantly reduced both gastric emptying and gastrointestinal tract transit. Such inhibitory effect was reduced by the C1 receptor antagonist SR141716A. Both morphological and pharmacological results suggest that FAAH may play a critical role in controlling gut anandamide levels.     

The urbanization of wildlife management: Social science, conflict, and decision making

Increasing urbanization of rural landscapes has created new challenges for wildlife management. In addition to changes in the physical landscape, urbanization has also produced changes in the socio-cultural landscape. The greater distancing from direct interaction with wildlife in urbanized societies has led to the emergence of a culture whose meanings for wildlife are less grounded in the utilitarian/instrumental orientation of rural agrarian systems. Urban perspectives on wildlife are comprised of more highly individualized emotional/symbolic values. This shift creates two problems with respect to managing wildlife in an urbanizing landscape. First the increased diversity in values and meanings increases the likelihood for social conflicts regarding wildlife management while at the same time making socially acceptable resolutions more intractable. This in turn requires fundamental changes in decision-making paradigms and the research approaches used to inform decision making. Second, as remaining rural communities feel the pressures of urbanization, wildlife conflicts become conflicts not just over wildlife but conflict over larger socio-political concepts such as equity, tradition, private property rights, government control, power, and acceptable forms of knowledge. This paper examines the wildlife management implications of changes associated with increasing urbanization and employs two case studies to illustrate these issues. First a study of a controversy over urban deer management provides insights into how to map conflicting values and search for common ground in an urban culture with increasingly individualistic values for wildlife. Specifically, the analysis illustrates that common ground may, at times, be found even among people with conflicting value systems. The second case study examined a ranching community faced with predator reintroduction. This case study illustrates tensions that occur when the community of interest (i.e. a national public) is broader than the community of place in which the problem occurs. In this latter situation, the debate centers around more than just different views about the rights of animals. It also entailed the rights of individuals and communities to decide their future. The conclusion discusses the need for wildlife institutions to adapt their underlying decision making philosophy including the way science is integrated into decision making processes in light of the changes in social context caused by urbanization.     

Assessing the potential impacts of alternative landscape designs on amphibian population dynamics

An individual-based, spatially explicit population model was used to predict the consequences of future land-use alternatives for populations of four amphibian species in two central Iowa (midwest USA) agricultural watersheds. The model included both breeding and upland habitat and incorporated effects of climatic variation and demographic stochasticity. Data requirements of the model include life history characteristics, dispersal behavior, habitat affinities, as well as land use and landcover in geographic information systems databases. Future scenarios were ranked according to change in breeder abundance, saturation, and distribution, compared to baseline conditions. Sensitivity of simulation results to changes in model parameters was also examined. Simulated results suggest that while all four species modeled are likely to persist under present and future scenario conditions, two may be more at risk from future landscape change. Although the study species are all widespread generalists regarded as having a low conservation priority, they depend on wetlands and ponds, increasingly endangered habitats in agricultural landscapes. Broader conservation strategies in the region would ensure that these currently common organisms do not become the endangered species of the future.This revised version was published online in May 2005 with corrections to the Cover Date.     

Shading effects on spatial growth and biomass partitioning of Cynodon dactylon

Cynodon dactylon is a low-growing C4 weed that is highly sensitive to shading. This species shows severe biomass reductions and highly plastic morphological changes in response to resource availability. The objective of this study was to analyse the effect of shading on the spatial and biomass growth of C. dactylon . Two experiments were carried out exploring different environmental conditions. Six treatments were applied on C. dactylon patches. Daylight and 18%, 41%, 48%, 69% and 85% shading of daylight photosynthetic active radiation (PAR) were imposed in Expt 1 and 0%, 22%, 44%, 50%, 74% and 87% shading of daylight PAR in Expt 2. Patches were slightly elliptical in shape and grew 3.2 and 1.5 cm d⁻¹ with 0% shading, on average, during the whole studied periods in Expts 1 and 2 respectively. Biomass was significantly reduced from 41% shading onwards in Expt 1 and from 50% shading in Expt 2. However, patch extension rate was only significantly reduced from 85% and 74% shading in Expts 1 and 2 respectively. Biomass partitioning was modified by shading in both experiments. A functional analysis of C. dactylon patches showed that the patch extension rate diminished linearly when patch biomass growth rate was lower than 1.66 g d⁻¹; above this value, the extension rate remained constant.     

Influence of the physical and chemical properties of soil on the retention process of isoxaflutole and its two main derivatives

Isoxaflutole is a new pre‐emergence herbicide for use in maize and sugarcane. Its two main derivatives are a diketonitrile derivative, the 2‐cyano‐3‐cyclopropyl‐1‐(2‐methanesulfonyl‐4‐trifluoromethylphenyl)propan‐1,3‐dione, named DKN, and a benzoic acid derivative, the 2‐methanesulfonyl‐4‐trifluoromethylbenzoic acid, named BA. The adsorption/desorption processes have never been studied for isoxaflutole (IFT) at high concentrations nor for BA, and the present work aimed at completing the knowledge of the behaviour of these three molecules in conditions close to those encountered in the context of agricultural use. The adsorption/desorption study was conducted on seven soils of different physical and chemical properties, using the batch equilibrium technique. During the experiments, IFT was chemically converted into DKN in a continuous manner. This reaction appeared to be dependent on the pH of the soil and was taken into account in the calculations of the adsorbed and desorbed amounts. The adsorption isotherms obtained were predominantly C‐shaped for IFT and DKN and S‐shaped for BA, but some differences appeared on a few soils. They fitted well the Freundlich equation, and the values of the Freundlich coefficient K_fa showed that, whatever the soil, IFT was more adsorbed than its two derivatives. The main parameter influencing the adsorption of IFT appeared to be the organic matter content, whereas this effect was not evident for DKN and BA. No correlation was found between the extent of adsorption and either clay content or pH of the soil, for the three molecules.     

Effects of winter wheat cultivars and seed rate on the biological characteristics of naturally occurring weed flora

Summary Differential competitive ability of six winter wheat cultivars and traits that confer such attributes were investigated for a range of seed rates in the presence or absence of weeds for a naturally occurring weed flora in two successive years in split-plot field experiments. Crop height and tillering capacity were considered suitable attributes for weed suppression, although competitiveness is a relative rather than an absolute characteristic. Maris Huntsman and Maris Widgeon were the most competitive cultivars whereas Fresco was the least competitive. Manipulation of seed rate was a more reliable factor than cultivar selection for enhancement of weed suppression, although competitiveness of cultivars Buster, Riband and Maris Widgeon was not enhanced by increased seed rate. Crop densities ranging between 125 and 270 plants m⁻² were found to offer adequate weed suppression. Linear relationships were observed between individual and total weed species dry weight and reproductive structures per unit area.     

Variation in the development of secondary dormancy in oilseed rape genotypes under conditions of stress

Summary A substantial amount of seed is left in the fields before and during harvest of oilseed rape. Although this crop exhibits little or no primary dormancy, the absence of certain environmental cues that promote germination of imbibed seeds induces secondary dormancy. The work reported investigated the extent to which environmental stress conditions, including osmotic stress, low oxygen stress and anaerobiosis, induce secondary dormancy in oilseed rape, and examined the variation in development of secondary dormancy between and within genotypes. Osmotic stress was most effective in inducing dormancy. Anaerobic treatment produced very few dormant seeds, as did an atmosphere low in oxygen and high in nitrogen. The development of secondary dormancy under osmotic stress varied considerably between and within genotypes. Dormancy ranged from almost zero to about 60% for winter genotypes and about 85% for spring types. Within genotypes, variations occurred between seed lots and years of harvest. Temperature variations affected the percentage of dormant seeds. More dormant seeds were likely to be produced with incubation under water stress at 20 °C than at 12 °C. In winter genotypes, fewer dormant seeds were produced when incubation temperature and germination test temperatures differed. Thus, incubating at 20 °C and 12 °C, followed by germination tests at 20 °C and 12 °C, respectively, produced most dormant seeds. Also, in the winter genotypes, the potential development of secondary dormancy was positively correlated with the pattern and speed of germination of untreated seeds.