Fixation requirements for the immunohistochemical reactivity of PCNA antibody PC10 on cryostat sections

Summary In contrast to that in paraffin-embedded tissue, the reactivity of monoclonal PCNA antibody PC10 on cryostat sections requires a special fixation procedure as the target epitope is seemingly not accessible to its antibody. A panel of 18 fixation protocols was investigated. Chilled methanol or acetone, or PLP (paraformaldehyde-lysine-periodate) was found to be unsuitable for skin preparations. A two-step fixation protocol was developed for normal skin and basal cell carcinomas. They were fixed first in 3.4% buffered formaldehyde, followed by fixation in 2:1 v/v ethanol-acetic acid. Following this fixation regime, cryostat sections displayed the same PCNA/PC10 labelling pattern as paraffin sections of formalin-fixed tissue.     

Paleolirnnological records of carotenoids and carbonaceous particles in sediments of some lakes in Southern Alps

Stratigraphic analyses of organic carbon, organic nitrogen and algal and bacterial carotenoids in short cores of profundal sediments of four alpine lakes (Tovel, Leit, Paione superiore and Tom) were used to reconstruct their trophic history. In addition, depth distribution of carbonaceous particle concentrations provided information on lake contamination from atmospheric deposition. In three lakes (Tovel, Leit and Tom), sedimentary carotenoids unique to sulfur photosynthetic bacteria (okenone and isorenieratene) provide evidence of changes in the oxygen, light and sulfide conditions in the water column. All the lakes are oligotrophic or moderately productive, and the algal community is dominated by Chlorophyta, Pyrrhophyta and Cryptophyta. Cyanobacteria are rather poorly represented. The steep increase of carbonaceous particles in the uppermost sediment layers of all the lakes suggests that lake contamination by atmospheric transport of pollutants began in the 1940s to 1950s. These data, coupled with those from a parallel study on Chrysophycean scale-inferred pH, indicate recent acidification in those which are poorly buffered (Paione superiore and Leit).     

Ein Vogelruf, dessen Form sich im Jahresablauf ?ndert: der Verbeugungstriller bei der Brandente (Tadorna tadorna)

Zusammenfassung Der Verbeugungstriller männlicher Brandenten, eine epigame Lautäußerung, weist sowohl in syntaktischen wie auch phonetischen Parametern zyklische jahreszeitliche Veränderungen auf. Die Gesamtdauer der Lautäußerung vergrößert sich allmählich im Laufe des Frühjahrs. Dafür sind eine Verlängerung des tonalen Eingangselements und eine Vermehrung der Elemente der abschließenden Phrase verantwortlich. Dagegen verkürzt sich das 2. Element gleichzeitig. In allen untersuchten Elementen erhöht sich die Tonhöhe parallel zur Verlängerung des Rufes. Diese Veränderungen des Ausdrucksverhaltens geben vermutlich Änderungen innerer Zustandsgrößen wieder.

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Seasonally changing bird call: the trill call of male Shelducks (Tadorna tadorna)

Both syntactic and phonetic features of the trill call accompanying the whistle-shake in adult male Shelducks are subject to cyclic annual changes. In the course of spring, the duration of the whole call increases gradually. This is caused by a prolongation of the first call element and insertion of additive elements in the final phrase. In contrast the second element is shortened by about 10 ms. At the same time, the pitch of all measured elements is raised. These findings are discussed in the framework of hormonal regulation and communicative function.

     

Patterns of cellular proliferation and migration in the developing tectum mesencephali of the frog Rana temporaria and the salamander Pleurodeles waltl

The development of the tectum mesencephali was studied in the frog Rana temporaria and the salamander Pleurodeles waltl by means of nuclear staining and by labeling of cells with bromodeoxyuridine (BrdU). The general spatial and temporal pattern of cell proliferation and cell migration is the same in both species, despite drastic differences in overall tectal morphology. However, the salamander species differs from the frog species by (1) a generally lower cell proliferation rate, (2) a reduction in the activity of the lateral proliferation zone, and (3) a reduction in the formation of superficial cellular layers. Because point (3) affects processes that occur late in ontogeny, our experiments provide evidence that the simple morphology of the tectum of Pleurodeles waltl, compared with the multilayered tectum of Rana, is a consequence of a paedomorphic alteration of the ancestral developmental pattern of the amphibian tectum.     

The distribution of coastal fish eDNA sequences in the Anthropocene

Aim

Coastal fishes have a fundamental role in marine ecosystem functioning and contributions to people, but face increasing threats due to climate change, habitat degradation and overexploitation. The extent to which human pressures are impacting coastal fish biodiversity in comparison with geographic and environmental factors at large spatial scale is still under scrutiny. Here, we took advantage of environmental DNA (eDNA) metabarcoding to investigate the relationship between fish biodiversity, including taxonomic and genetic components, and environmental but also socio-economic factors.

Location

Tropical, temperate and polar coastal areas.

Time period

Present day.

Major taxa studied

Marine fishes.

Methods

We analysed fish eDNA in 263 stations (samples) in 68 sites distributed across polar, temperate and tropical regions. We modelled the effect of environmental, geographic and socio-economic factors on α- and β-diversity. We then computed the partial effect of each factor on several fish biodiversity components using taxonomic molecular units (MOTU) and genetic sequences. We also investigated the relationship between fish genetic α- and β-diversity measured from our barcodes, and phylogenetic but also functional diversity.

Results

We show that fish eDNA MOTU and sequence α- and β-diversity have the strongest correlation with environmental factors on coastal ecosystems worldwide. However, our models also reveal a negative correlation between biodiversity and human dependence on marine ecosystems. In areas with high dependence, diversity of all fish, cryptobenthic fish and large fish MOTUs declined steeply. Finally, we show that a sequence diversity index, accounting for genetic distance between pairs of MOTUs, within and between communities, is a reliable proxy of phylogenetic and functional diversity.

Main conclusions

Together, our results demonstrate that short eDNA sequences can be used to assess climate and direct human impacts on marine biodiversity at large scale in the Anthropocene and can further be extended to investigate biodiversity in its phylogenetic and functional dimensions.     

Novel Miscanthus hybrids: Modelling productivity on marginal land in Europe using dynamics of canopy development determined by light interception

New biomass crop hybrids for bioeconomic expansion require yield projections to determine their potential for strategic land use planning in the face of global challenges. Our biomass growth simulation incorporates radiation interception and conversion efficiency. Models often use leaf area to predict interception which is demanding to determine accurately, so instead we use low-cost rapid light interception measurements using a simple laboratory-made line ceptometer and relate the dynamics of canopy closure to thermal time, and to measurements of biomass. We apply the model to project the European biomass potentials of new market-ready hybrids for 2020–2030. Field measurements are easier to collect, the calibration is seasonally dynamic and reduces influence of weather variation between field sites. The model obtained is conservative, being calibrated by crops of varying establishment and varying maturity on less productive (marginal) land. This results in conservative projections of miscanthus hybrids for 2020–2030 based on 10% land use conversion of the least (productive) grassland and arable for farm diversification, which show a European potential of 80.7–89.7 Mt year⁻¹ biomass, with potential for 1.2–1.3 EJ year⁻¹ energy and 36.3–40.3 Mt year⁻¹ carbon capture, with seeded Miscanthus sacchariflorus × sinensis displaying highest yield potential. Simulated biomass projections must be viewed in light of the field measurements on less productive land with high soil water deficits. We are attempting to model the results from an ambitious and novel project combining new hybrids across Europe with agronomy which has not been perfected on less productive sites. Nevertheless, at the time of energy sourcing issues, seed-propagated miscanthus hybrids for the upscaled provision of bioenergy offer an alternative source of renewable energy. If European countries provide incentives for growers to invest, seeded hybrids can improve product availability and biomass yields over the current commercial miscanthus variety.     

Attempts to Determine the Roles of Visual and Olfactory Inputs in Initial Orientation and Homing of Pigeons over Familiar Terrain

The role of familiar visual landmarks in pigeon homing is still unsettled. If they are involved, they must be thought to be utilized in parallel with olfactory signals. In order to recognize the effectiveness of either one of the input channels separately, vision and olfaction, it is therefore necessary to interfere with both of them. Pigeons were temporarily deprived of image vision by spectacles made of translucent white paper, producing a condition called V^- as compared with the largely unimpaired condition V⁺. Access to olfactory signals was temporarily prohibited by charcoal filters before release and nasal anaesthesia upon release, resulting in condition O^- versus unimpaired smelling of natural air in O⁺. Prior to the test releases, all the participating pigeons had been made familiar with the two or four test sites (and with other sites in the area) by training flights. According to decreasing levels of initial homeward orientation and homing performance, the four combinations of treatments are to be arranged in the following order: V⁺O⁺, V⁺O^-, V^-O⁺, V^-O^-. In the last type, no trace of initial homeward orientation remained, indicating that at least one of the two input channels need be functional to enable home-related orientation. Over well known terrain, either alone seems to be more or less sufficient. The results are indicative but not definitely conclusive. Effects of visual impairment on behavioural activities in general cannot clearly be separated from orientation-specific effects. Therefore, and because many pigeons refuse to fly in V^- condition or fail to provide useful data, we stopped applying this method.