Two hundred years of land-use change in the South Swedish Uplands: comparison of historical map-based estimates with a pollen-based reconstruction using the landscape reconstruction algorithm

Long-term records of environmental history at decadal to millennial time-scales enable an assessment of ecosystem variability and responses to past anthropogenic disturbances and are fundamental for the development of environmental management strategies. This study examines the local variability of land-use history in the South Swedish Uplands over the last 200 years based on pollen records from three lake-sediment successions. Temporal changes in the proportional cover of 14 plant taxa were quantified as percentages using the landscape reconstruction algorithm (LRA). The LRA-based estimates of the extent of four land-use categories (cropland, meadows/grassland, wetland, outland/woodland) were compared to corresponding estimates based on historical maps and aerial photographs from ad 1769–1823, 1837–1895, 1946 and 2005. Although the LRA approach tends to overestimate grassland cover by 10–30 % for the two earliest time periods, the reconstructed vegetation composition is generally in good agreement with estimates based on the historical records. Subsequently, the LRA approach was used to reconstruct the 200-year history of local land-use dynamics at 20-year intervals around two small lakes. The qualitative assessment of difference approach, which requires fewer assumptions and parameters than LRA for objective evaluation of between-site differences in plant abundances, provides consistent results in general. Significant differences exist in the land-use history between the sites. Local catchment characteristics, such as soil conditions and wetland cover, appear important for the development of human impact on the landscape. Quantifications of past vegetation dynamics provide information on the amplitude, frequency and duration of the land-use changes and their effects on terrestrial and aquatic ecosystems, and should be taken into account when nature conservation strategies are developed.     

Asynchronous population dynamics of Siberian lemmings across the Palaearctic tundra

The synchrony of Siberian lemming (Lemmus sibiricus L.) population dynamics was investigated during a ship-borne expedition along the Palaearctic tundra coast in the summer of 1994. On 12 sites along the coast from the Kola Peninsula to Wrangel Island, relative densities of lemmings were recorded using a standardised snap-trapping programme. The phase position of the lemming cycle in each of the studied populations was determined based on current density estimates, signs of previous density and the age profile of each population (ageing based on eye lens mass). In addition, dendrochronological methods were used to determine when the last peak in the density of microtine populations occurred at each site. The examined lemming populations were in different phases of the lemming cycle. Some populations were in the peak phase, as indicated by high current densities, an age profile in which older individuals were well represented, and signs of high previous density (abundant old lemming faeces). Other populations were in the decline phase, as reflected in a moderate current density, a predominance of older individuals and signs of high previous density. Populations in the low phase had an extremely low current density and showed signs of high previous density, while populations in the increase phase had a moderate current density, a predominance of younger individuals and showed signs of low previous density. The results of phase determinations based on dendrochronological methods support the findings based on lemming demography. Recent Russian studies carried out on some of the sites also agreed with our phase determination results. Thus, on a regional scale (across the whole Palaearctic tundra), the population dynamics of Siberian lemmings can be considered asynchronous. However, sites situated adjacent to each other were often phase synchronous, suggesting a more fine-grained pattern of dynamics with synchrony over distances as long as 1000 km or so, e.g. the Yamal and Taymyr Peninsulas. Received: 19 August 1998 / Accepted: 1 March 1999     

The folding and stability of human alpha class glutathione transferase A1-1 depend on distinct roles of a conserved N-capping box and hydrophobic staple motif.

An N-capping box and a hydrophobic staple motif are strictly conserved in the core of all known glutathione S-transferases (GST). In the present work, mutations of hGSTA1-1 enzyme residues forming these motifs have been generated. The analysis of S154A, D157A, and S154A/D157A capping mutants indicate that the removal of this local signal destabilizes the protein. The fact that the third helical residue D157A mutation (N-3) was much more destabilizing than the first helical residue S154A mutation (N-cap) suggests that the appropriate conformation of the conserved substructure formed by the alpha 6-helix and preceding loop (GST motif II) is crucial for the overall protein stability. The refolding study of GSTA1-1 variants supports the prediction that this subdomain could represent a nucleation site of refolding. The analysis of L153A, I158A, L153G, and L153A/I158A hydrophobic staple mutants indicate that the removal of this motif destabilizes the GSTA1-1 structure as well as its refolding transition state. The hydrophobic staple interaction favors essential inter-domain contacts and, thereby, in contrast to capping interactions, accelerates the enzyme reactivation. Its strict conservation in the GST system supports the suggestion that this local signal could represent an evolutionarily conserved determinant for rapid folding.     

Titration and subunit localization of active center cysteine in fibrinoligase (thrombin-activated fibrin stabilizing fector)

Fibrinoligase activity, measured in a fully synthetic substrate system, was shown to be inhibited by prior incubation of the enzyme with iodoacetamide. Using 1-¹⁴C-iodoacetamide, the amount of incorporated radio-activity increased in proportion to the loss of enzyme activity. Labelling of the enzyme was totally dependent on the addition of calcium ions and, under the conditions described, occurred predominantly in the $\text{a}\text{′}$ protomer. Acid hydrolysis of labelled fibrinoligase gave rise to S-carboxymethylcysteine as the only radioactive amino acid derivative.     

Porous gelatin hydrogels: 1. Cryogenic formation and structure analysis

In the present work, porous gelatin scaffolds were prepared by cryogenic treatment of a chemically cross-linked gelatin hydrogel, followed by removal of the ice crystals formed through lyophilization. This technique often leads to porous gels with a less porous skin. A simple method has been developed to solve this problem. The present study demonstrates that the hydrogel pore size decreased with an increasing gelatin concentration and with an increasing cooling rate of the gelatin hydrogel. Variation of the cryogenic parameters applied also enabled us to develop scaffolds with different pore morphologies (spherical versus transversal channel-like pores). In our opinion, this is the first paper in which temperature gradients during controlled cryogenic treatment were applied to induce a pore size gradient in gelatin hydrogels. With a newly designed cryo-unit, temperature gradients of 10 and 30 degrees C were implemented during the freezing step, resulting in scaffolds with average pore diameters of, respectively, +/-116 and +/-330 microm. In both cases, the porosity and pore size decreased gradually through the scaffolds. Pore size and structure analysis of the matrices was accomplished through a combination of microcomputed tomography using different software packages (microCTanalySIS and Octopus), scanning electron microscopy analysis, and helium pycnometry.     

Establishment of HSV1 Latency in Immunodeficient Mice Facilitates Efficient In Vivo Reactivation

The establishment of latent infections in sensory neurons is a remarkably effective immune evasion strategy that accounts for the widespread dissemination of life long Herpes Simplex Virus type 1 (HSV1) infections in humans. Periodic reactivation of latent virus results in asymptomatic shedding and transmission of HSV1 or recurrent disease that is usually mild but can be severe. An in-depth understanding of the mechanisms regulating the maintenance of latency and reactivation are essential for developing new approaches to block reactivation. However, the lack of a reliable mouse model that supports efficient in vivo reactivation (IVR) resulting in production of infectious HSV1 and/or disease has hampered progress. Since HSV1 reactivation is enhanced in immunosuppressed hosts, we exploited the antiviral and immunomodulatory activities of IVIG (intravenous immunoglobulins) to promote survival of latently infected immunodeficient Rag mice. Latently infected Rag mice derived by high dose (HD), but not low dose (LD), HSV1 inoculation exhibited spontaneous reactivation. Following hyperthermia stress (HS), the majority of HD inoculated mice developed HSV1 encephalitis (HSE) rapidly and synchronously, whereas for LD inoculated mice reactivated HSV1 persisted only transiently in trigeminal ganglia (Tg). T cells, but not B cells, were required to suppress spontaneous reactivation in HD inoculated latently infected mice. Transfer of HSV1 memory but not OVA specific or naïve T cells prior to HS blocked IVR, revealing the utility of this powerful Rag latency model for studying immune mechanisms involved in control of reactivation. Crossing Rag mice to various knockout strains and infecting them with wild type or mutant HSV1 strains is expected to provide novel insights into the role of specific cellular and viral genes in reactivation, thereby facilitating identification of new targets with the potential to block reactivation.     

Deficit of CD47 results in a defect of marginal zone dendritic cells, blunted immune response to particulate antigen and impairment of skin dendritic cell migration

CD47 is a ubiquitously expressed cell surface glycoprotein that associates with integrins and regulates chemotaxis, migration, and activation of leukocytes. CD47 is also a ligand for signal regulatory protein alpha, a cell surface receptor expressed on monocytes, macrophages, granulocytes, and dendritic cell (DC) subsets that regulates cell activation, adhesion, and migration. Although the function of CD47 in macrophages and granulocytes has been studied in detail, little is known about the role of CD47 in DC biology in vivo. In this study we demonstrate that CD47(-/-) mice exhibit a selective reduction of splenic CD11c(high)CD11b(high)CD8alpha(-)CD4(+) DCs. These DCs correspond to marginal zone DCs and express signal regulatory protein alpha, possibly explaining their selective deficiency in CD47(-/-) mice. Deficiency of marginal zone DCs resulted in impairment of IgG responses to corpusculate T cell-independent Ags. Although epidermal DCs were present in normal numbers in CD47(-/-) mice, their migration to draining lymph nodes in response to contact sensitization was impaired, while their maturation was intact. In vitro, CD47(-/-) mature DCs showed normal CCR7 expression but impaired migration to CCL-19, whereas immature DC response to CCL-5 was only slightly impaired. These results demonstrate a fundamental role of CD47 in DC migration in vivo and in vitro and in the function of marginal zone DCs.     

Direct monitoring of vesicular release and uptake in brain slices by multiphoton excitation of the styryl FM 1-43

Fluorescence imaging using FM 1-43 and related styryl dyes has provided invaluable insights into presynaptic function of synapses in culture preparations, but has been limited in use for studying central synapses in vivo or in brain slices, because of excessive fluorescence background due to nonspecific membrane binding of dye. We demonstrate here that focal excitation of FM dyes using two-photon laser-scanning microscopy (TPLSM) provides high resolution of FM 1-43-labeled nerve terminals in brain slices by suppressing out-of-focus background and that a readily releasable pool of vesicles can be selectively and stably labeled by hypertonic shock despite slice diffusion barriers. We find direct TPLSM of FM 1-43-labeled nerve terminals to be superior to treatment of slices with either the fluorescent quencher sulforhodamine 101 or dye scavenger ADVASEP-7 in resolving nerve terminal against background fluorescence, enabling continuous monitoring of vesicular uptake, and release of styryl dyes from individual nerve terminals in brain slices.     

Soil Resources Influence Spatial Patterns of Denitrifying Communities at Scales Compatible with Land Management

Knowing spatial patterns of functional microbial guilds can increase our understanding of the relationships between microbial community ecology and ecosystem functions. Using geostatistical modeling to map spatial patterns, we explored the distribution of the community structure, size, and activity of one functional group in N cycling, the denitrifiers, in relation to 23 soil parameters over a 44-ha farm divided into one organic and one integrated crop production system. The denitrifiers were targeted by the nirS and nirK genes that encode the two mutually exclusive types of nitrite reductases, the cd₁ heme-type and copper reductases, respectively. The spatial pattern of the denitrification activity genes was reflected by the maps of the abundances of nir genes. For the community structure, only the maps of the nirS community were related to the activity. The activity was correlated with nitrate and dissolved organic nitrogen and carbon, whereas the gene pools for denitrification, in terms of size and composition, were influenced by the soil structure. For the nirS community, pH and soil nutrients were also important in shaping the community. The only unique parameter related to the nirK community was the soil Cu content. However, the spatial pattern of the nirK denitrifiers corresponded to the division of the farm into the two cropping systems. The different community patterns, together with the spatial distribution of the nirS/nirK abundance ratio, suggest habitat selection on the nirS- and nirK-type denitrifiers. Our findings constitute a first step in identifying niches for denitrifiers at scales relevant to land management.Soil microorganisms are abundant and diverse (46), drive key processes in biogeochemical cycles, and, thus, play crucial roles in ecosystem functioning (2). They are not randomly distributed but exhibit spatial patterns at different scales (26). Spatial patterns ranging from the micrometer up to the meter scale have been reported (19, 20, 32, 37), and an understanding of such patterns can give clues to how microbial communities are generated and maintained (17). Spatial patterns of microorganisms at the field and landscape scales warrant special attention, since they could be associated with land use and aid in creating knowledge-based management strategies for agricultural production (5, 42). However, our understanding of key habitat-selective factors is limited, and few studies have specified which factors influence the spatial patterns of soil microbial communities at larger scales. Lauber et al. (30) recently demonstrated that pH could predict the community composition of soil bacteria at the continental scale. The importance of pH as a key edaphic driver of bacterial community structure has also been shown in other studies (11, 47). Another major, but complex, factor pointed out in a few studies is the soil type (4, 5, 14). Most studies have included only a limited number of properties that are easy to measure; most often, carbon and nitrogen pools and soil physical factors have been neglected in microbial community ecology. Since these factors delineate soil oxygen and water content, they may exert a stronger impact on microbial communities than other soil resources.Reports on the field or landscape scale spatial distribution of soil bacteria have had a taxon-centered perspective at either the species or total-community level, but there is emerging interest in the biogeography of functional traits possessed by microorganisms (18). Bacterial species composition is likely important for soil ecosystem functions, but species affiliation rarely predicts in which way. In addition, the fuzzy species concept of bacteria makes it all the more difficult to link species to niches. Analysis of functional guilds, i.e., assemblages of populations sharing certain traits, can bridge this gap, and one guild of global concern that has been suggested and recently used as a model in functional ecology is the denitrifiers (39, 49). Denitrification is an anaerobic respiration pathway during which NO₃⁻ is reduced to N₂ by a wide range of unrelated taxa. The process is an essential route for N loss from agricultural soil and a major source of the greenhouse gas N₂O. It was recently shown that the spatial distribution of the relative abundances of denitrifiers with the genetic capacity to perform the last step in the denitrification pathway, reduction of N₂O to N₂, is linked to areas with high denitrification rates and low N₂O emissions (38). Adding field scale predicted patterns of the denitrifier community structure to the abundance and activity would not only give insight into the mechanisms shaping the community, but also deepen our understanding of the relationships between the ecology of denitrifiers, N loss, and the agroecosystems'' impact on climate change.We hypothesize that spatial autocorrelations of the structures, sizes, and activities of communities of denitrifying bacteria is governed by soil-based resources at a scale compatible with land management. To test this, and to elucidate the effects of crop production systems and the importance of soil physical and chemical factors in the denitrifying community, we explored the spatial distribution of community structure, size, and activity in relation to 23 soil parameters at a 44-ha farm divided into one organic and one integrated crop production system. The denitrifier community was described in terms of the signature genes that encode the two different types of nitrite reductases in the denitrification pathway, the cd₁ heme-type reductase (NirS), encoded by the nirS gene, and the copper oxidoreductase (NirK), encoded by nirK. The spatial patterns were mapped by geostatistical modeling, and correlation structures were explored.     

Identification of Key Residues Determining Species Differences in Inhibitor Binding of Microsomal Prostaglandin E Synthase-1

Microsomal prostaglandin E synthase-1 (MPGES1) is induced during an inflammatory reaction from low basal levels by pro-inflammatory cytokines and subsequently involved in the production of the important mediator of inflammation, prostaglandin E₂. Nonsteroidal anti-inflammatory drugs prevent prostaglandin E₂ production by inhibiting the upstream enzymes cyclooxygenases 1 and 2. In contrast to these conventional drugs, a new generation of NSAIDs targets the terminal enzyme MPGES1. Some of these compounds potently inhibit human MPGES1 but do not have an effect on the rat orthologue. We investigated this interspecies difference in a rat/human chimeric form of the enzyme as well as in several mutants and identified key residues Thr-131, Leu-135, and Ala-138 in human MPGES1, which play a crucial role as gate keepers for the active site of MPGES1. These residues are situated in transmembrane helix 4, lining the entrance to the cleft between two subunits in the protein trimer, and regulate access of the inhibitor in the rat enzyme. Exchange toward the human residues in rat MPGES1 was accompanied with a gain of inhibitor activity, whereas exchange in human MPGES1 toward the residues found in rat abrogated inhibitor activity. Our data give evidence for the location of the active site at the interface between subunits in the homotrimeric enzyme and suggest a model of how the natural substrate PGH₂, or competitive inhibitors of MPGES1, enter the active site via the phospholipid bilayer of the membrane.