Isolation and characterization of bone marrow-derived equine mesenchymal stem cells

OBJECTIVE: To isolate and characterize bone marrow-derived equine mesenchymal stem cells (MSCs) for possible future therapeutic applications in horses. SAMPLE POPULATION: Equine MSCs were isolated from bone marrow aspirates obtained from the sternum of 30 donor horses. PROCEDURES: Cells were cultured in medium (alpha-minimum essential medium) with a fetal calf serum content of 20%. Equine MSC features were analyzed to determine selfrenewing and differentiation capacity. For potential therapeutic applications, the migratory potential of equine MSCs was determined. An adenoviral vector was used to determine the transduction rate of equine MSCs. RESULTS: Equine MSCs can be culture-expanded. Equine MSCs undergo cryopreservation in liquid nitrogen without altering morphologic characteristics. Furthermore, equine MSCs maintain their ability to proliferate and differentiate after thawing. Immunocytochemically, the expression of the stem cell marker CD90 can be detected on equine MSCs. The multilineage differentiation potential of equine MSCs was revealed by their ability to undergo adipogenic, osteogenic, and chondrogenic differentiation. CONCLUSIONS AND CLINICAL RELEVANCE: Our data indicate that bone marrow-derived stromal cells of horses can be characterized as MSCs. Equine MSCs have a high transduction rate and migratory potential and adapt to scaffold material in culture. As an autologous cell population, equine MSCs can be regarded as a promising cell population for tissue engineering in lesions of the musculoskeletal system in horses.     

Relationship between somaclonal variation and type of culture in cucumber

Highly inbred B line of cucumber was used to compare the effect of four types of in vitro culture on somaclonal variation. The plants were regenerated from the following types of culture: twelve- and eighteen-month-old liquid culture of meristematic clumps (LMC₁₂₍₁₈₎), ten-month-old embryogenic cytokinin-dependent suspension (CDS), eighteen-month-old embryogenic cytokinin-dependent suspension in medium with modified NH⁺ ₄/NO₃ ^- ratio (CDS 1.7), twelve-month-old embryogenic auxin-dependent suspension (ADS), thirty six-month-old embryogenic auxin-dependent suspension in medium with modified NH⁺ ₄/NO₃ ^- ratio (ADS 1.7) and recurrent leaf callus regeneration (RLC) – repeated 5 times. The differences in the incidence of the following properties were observed: the ploidy of R₀ plants, the segregation of new morphological traits in R₁ and the germination ability of R₁ seeds. R₁ families with the segregation of new phenotypes were most numerous in CDS (62.5%) and LMC₁₈ (57.9%), next in CDS1.7 (35.7%), while the smallest number was found in LMC₁₂ (11.1%) and RLC (3.4%).Tetraploid and mixoploid plants occurred in ADS1.7 and ADS (100%) whereas CDS and RLC were observed to contain only tetraploids, respectively 33.3% and 55.2%. There were no changes of ploidy after LMC₁₂, LMC₁₈ and CDS1.7. Among new phenotypes there were such that have not been described so far in cucumber: ginkgolike leaf (gll), yellow-green chlorophyll mutants (y-gc), serrate margin of corolla in male and female flowers (smc). This revised version was published online in August 2006 with corrections to the Cover Date.     

Long-term impact of fertilization on activity and composition of bacterial communities and metabolic guilds in agricultural soil

To explore long-term impact of organic and inorganic fertilizers on microbial communities, we targeted both the total bacterial community and the autotrophic ammonia oxidizing bacteria (AOB) in soil from six treatments at an experimental field site established in 1956: cattle manure, sewage sludge, Ca(NO₃)₂, (NH₄)₂SO₄, unfertilized and unfertilized without crops. All plots, except the bare fallows, were cropped with maize. Effects on activity were assessed by measuring the basal respiration and substrate induced respiration (SIR) rates, and the potential activity of the AOB. To determine the bacterial community composition, 16S rRNA genes were used to fingerprint total soil communities by terminal restriction fragment length polymorphism analysis and AOB communities by denaturing gradient gel electrophoresis. The fertilization regimes had clear effects on both activity and composition of the soil communities. Basal respiration and r, which was kinetically derived as the exponentially growing fraction of the SIR-response, correlated well with the soil organic C content (r=0.93 and 0.66, respectively). Soil pH ranged from 3.97 to 6.26 in the treatments and was found to be an important factor influencing all microbial activities. pH correlated negatively with the ratio between basal respiration and SIR (r=0.90), indicating a decreased efficiency of heterotrophic microorganisms to convert organic carbon into microbial biomass in the most acid soils with pH 3.97 and 4.68 ((NH₄)₂SO₄ and sewage sludge fertilized plots, respectively). The lowest SIR and ammonia oxidation rates were also found in these treatments. In addition, these treatments exhibited individually different community fingerprints, showing that pH affected the composition of AOB and total bacterial communities. The manure fertilized plots harbored the most diverse AOB community and the pattern was linked to a high potential ammonia oxidation activity. Thus, the AOB community composition appeared to be more strongly linked to the activity than the total bacterial communities were, likely explained by physiological differences in the populations present.     

Urbanization and homogenization - Comparing the floras of urban and rural areas in Germany

The process of urbanization has resulted in an expansion of alien plant species and declines of native species, in particular already rare species. These processes may cause a greater similarity between different urban regions, i.e. biotic homogenization. We explored the relationship between urban regions and homogenization for plant species in Germany using (i) all plant species, (ii) only native species, (iii) all alien species and only those introduced (iv) before or (v) after 1500, respectively. We used the database FLORKART for species distribution on a 6 min latitude × 10 min longitude (ca. 130 km²) scale. We calculated mean similarities for the 60 “most urbanized” cells. We then resampled 60 randomly drawn “less urbanized” cells and 60 “rural” cells and compared these results to the “most urbanized” cells taking distance effects into account. Urbanization does not have an overall effect on homogenization of all species, but native species as well as pre-1500 alien plant assemblages show effects of homogenization while post-1500 alien plant assemblages show the opposite effect. On a regional scale, urbanization is not unequivocally related to homogenization. This might be different when extending the analysed range across several bioclimatic regions. Specific urban habitats, or what remains of them, require special protection and management if the trend towards homogenization is to be avoided.     

Embolism induced by winter drought may be critical for the survival of <Emphasis Type="Italic">Pinus sylvestris</Emphasis> L. near its southern distribution limit

Introduction   

Scots pine populations in the SE “Sistema Ibérico” range suffered a severe defoliation in the S face of the crown in isolated trees of thinned stands. This process was detected at the end of the winter 2001–2002.     

Review of ground-based methods to measure the distribution of biomass in forest canopies

? Context

Ecological research and an effective forest management need accurate information on the structure of the forest canopy to understand the biochemical, physiological and biogeochemical processes within a forest.

? Research question

This paper reviews the currently available instruments for measuring the distribution of biomass within forest canopies. We compare the most well-established approaches and present the different measurable parameters. A special focus lies on the resolution of the obtained data.

? Results

It was found that only 3D laser scanners offer data with the resolution required by ecologists, private landholders, the forest industry and the public to detect trends in tree growth patterns and canopy interactions in all three spatial dimensions. But data validation, data analysis and parameter extraction are still under development, and the price of the instrument is quite high.

? Conclusion

Research should focus on the parameter extraction from terrestrial laser scanner data as this could allow the calculation of functional attributes for different sections of a canopy on a high spatial resolution. It could also help ecologists characterize the structure of forest stands in a quick and precise way.     

Speciation in the parthenogenetic oribatid mite genus Tectocepheus (Acari, Oribatida) as indicated by molecular phylogeny

Despite the lack of universal concepts for species and speciation, both sexual and asexual organisms are expected to diversify into discrete genotypic and morphological clusters. Species-rich clusters of parthenogenetic oribatid mites likely evolved in the absence of sexual reproduction. We used nucleotide sequences of the large and small rDNA genes (18S and 28S) and the coding genes for heat shock protein 82 (hsp82) and elongation factor-1 alpha (ef-1α) for phylogenetic analyses of three morphotypes of the parthenogenetic oribatid mite genus Tectocepheus. DNA sequence divergences of the different morphotypes were similar to those of sexual species in other organisms. Maximum likelihood analyses of single genes and combined data sets were largely congruent in reconstructing the phylogeny of the Tectocepheus species. The results suggest that the different morphotypes of Tectocepheus indeed evolved in the absence of sexual reproduction, and that Tectocepheus minor, Tectocepheus velatus and Tectocepheus sarekensis are best considered distinct species. Further, the results suggest that 18S rDNA, hsp82 and ef-1α are powerful markers for resolving phylogenetic relationships of oribatid mites. Saturation plots indicated that the D3-region of 28S is much more saturated than all other examined genes. This indicates that the D3-region is unsuitable for resolving ancient splits in oribatid mites.     

Element (Ag, Cd, Cu, Pb, Sb, Tl and Zn), element ratio and lead isotope profiles in a sediment affected by a mining operation episode during the late 19th century

Mining operations at Mårsätter in 1877–81 resulted in increased metal loading to a small lake, notably as sulphidic tailings. The event is taken as an opportunity to study the present environmental impact of a historical single major metal release. Lake water and four sediment cores were sampled and analysed for principal and trace elements in solid and aqueous phases as well as general hydrochemical conditions. Chronologies were determined from ²⁰⁶Pb/²⁰⁷Pb ratios and historical records. Ordinary sedimentation after the event has lead to that the tailings are found as a distinct layer at a depth of 18–22 cm in the sediment. The layer is characterized by elevated metal concentrations in the solid and pore water phases, respectively, circum neutral pH and sulphate concentrations below detection. Geochemical modelling indicated a preference for carbonate equilibrium in the waste while sulphides prevailed above it. It is concluded that the growth of the ordinary sediment on top of the waste has lead to a physicochemical barrier that seals of the waste from the overlying sediment. Chemical or physical rupture of the barrier will release the metals to downstream regions. According to the chronologies at least three sources have contributed to the present elevated levels of metals, in additions to the release of tailings. Copper from a historical blast furnace prior to the event at Mårsätter, transport from mineralized parts of the watershed and release of contaminated water from present mining operations maintain elevated levels of notably zinc, silver, cadmium and lead. At present less than 10% of the lead content at the sediment/water interface comes from atmospheric deposition. Increased levels of antimony and thallium were not observed prior to ca 1950.     

Release of nonexchangeable NH4-N after planting of ryegrass in relation to soil content and as affected by nitrate supply

The uptake of N by ryegrass grown in pot culture on a range of soils differing widely in content of nonexchangeable NH₄-N (topsoils: 117 to 354 mg kg^?1 soil; subsoils: 117 to 270 mg kg^?1 soil) was measured to indicate whether the amounts of NH₄-N released from clay minerals were correlated with soil NH₄-N. After two cuts soil analysis revealed that the amounts of mobilized nonexchangeable NH₄-N were between 3.5 and 25.2 mg kg^?1 from topsoils and between 0 and 8.2 mg kg^?1 from subsoils. There was no correlation between soil nonexchangeable NH₄-N content and release. The NH₄-N extracted with 1 N HCl and the actual N uptake of the plants correlated highly significant. Assuming that the whole of the NH₄-N released was taken up by ryegrass, NH₄-N accounted for 11.2 to 75.0% of total N uptake from topsoils and 0 to 37.3% from subsoils. The release of nonexchangeable NH₄-N was increased by the application of nitrate.     

Little effect of forest age on oribatid mites on the bark of trees

This study investigates the effect of forest age (20-, 50-, 160-year-old, and primeval forest) on oribatid mite communities on the bark of oak trees in the National Park Pusza Białowieża in eastern Poland. We hypothesized that oribatid mite diversity on bark peaks at forests of intermediate age and that the number of parthenogenetic species of oribatid mites is highest in young stands. In contrast to these hypotheses, the diversity, density, number of juveniles, community structure and the mode of reproduction of oribatid mite species were not significantly affected by forest age. None of the oribatid mite species occurred exclusively on trees of a specific age. The results suggest that oribatid mite communities on the bark of trees are minimally affected by tree harvesting regimes. In contrast to oribatid mites in soil, communities on bark appear to be less sensitive to disturbances.