Vegetation survey in the former brown coal mining area of eastern Germany by integrating remote sensing and ground-based methods

Abstract. In the former brown coal mining area of eastern Germany, now scheduled as a nature conservation area, an analysis of the spatial distribution of vegetation was considered as an important tool in landscape planning. Therefore a comprehensive vegetation survey by means of satellite imagery (Landsat-TM), airborne imagery (CASI), and ground-based methods, notably habitat mapping and vegetation sampling was carried out. With respect to the scales of resolution the classification results of the four methods are, to a certain degree, comparable. Differences in the outcome can be ascribed to the fact that methods of low resolution result in a discrete array of polygons whereas methods of high resolution depict a mosaic structure with an underlying, continuously changing gradient. Provided that the biological meaning of the remote sensing classification is known, a shift from single vegetation patterns to the landscape scale will be possible. Neither satellite nor airborne imagery is restricted to the purpose of mapping but may also serve for vegetation classification itself.     

Quantum-dye labeled proteins for glycobiology: a viable nonradioactive alternative tracer

Quantum dye (QD), a macrocyclic europium-chelate, developed as a cytological marker, has never been used for quantitative applications. It would be ideal, however, if the same tracer can be used for both qualitative and quantitative purposes. We have labeled some lectins and neoglycoproteins with QD for the purpose of quantitative analyses in glycobiology, and tested its suitability in three different areas in glycobiology: (1) glycosyltransferase, (2) an animal lectin - mannose- binding protein, and (3) the Gal/GalNAc receptor of rat liver membrane. Usefulness of QD-labeled lectins was amply demonstrated by the quantification of galactosyltransferase activity using QD-soybean agglutinin and QD-RCA120 ( Ricinus communis agglutinin). We also showed that QD-labeled neoglycoproteins, QD-Man-BSA and QD-Gal-BSA, can replace radioiodinated counterparts in the binding assays of animal lectins (serum mannose binding protein and hepatic Gal/GalNAc receptor.) The advantage of QD and other europium labels is that it does not decay as radioiodides do. The long shelf-life results in more consistent results from repeated experiments.      

Assessment of the effect of alpha-galactosides injected during embryogenesis on selected chicken traits

The effect of different doses of alpha-galactoside (RFOs) preparations from Pisum sativum L. cv. Opal, injected into eggs during embryogenesis, on maintaining a high number of bifidobacteria, selected chicken broiler traits and the lipoprotein level of blood were studied. Two independent experiments were conducted. In the first, Ringer water solution containing 1.763 mg/egg of fructooligosaccharides (FOS) (I group), 2.1158 mg of pea RFO preparation containing 20% sucrose (II group) and 0.4232 mg of sucrose (III group) were injected into Hubbard broiler breeder eggs containing 12-day old embryos. Only Ringer water solution was applied to the eggs of the control group (IV group). The number of bifidobacteria determined in faeces of two-day old chicken of groups I and II was significantly higher in comparison with the sucrose and control groups. The high level ofbifidobacteria of groups I and II was maintained during 6 weeks. The dose of both preparations had no influence on the body weight, carcass, breast muscle, leg and abdominal fat ratio, total cholesterol, HDL and LDL serum concentrations. Broiler mortality and breast muscle cholesterol concentration was highest (P < 0.05) for the control group. On the other hand, the European Production Index, as well as serum triglycerides, were the lowest for this group. The second experiment was performed on Hybro G chicken breeder eggs. 0.69, 3.43 and 6.87 mg/egg of pea RFO preparation doses containing 20% sucrose were injected into the experimental groups. The number of bifidobacteria in the caecum and selected meat traits of broilers were determined. The results of this experiment confirmed that RFO injection in ovo causes the long-time maintenance of a high level ofbifidobacteria. The dose of the preparations does not have any effect on the selected broiler meat traits, except that the highest dose increases the percent of carcase in body weight. However, this dose reduced the hatchability of the treated embryos.     

Generation of cytotoxic T cells specific for minor histocompatibility antigens by cross challenge in vitro with H-2 disparate adherent cells

Although it is well known that an H-2-restricted cytotoxic T cell response to minor histocompatibility antigens (MIHA) can be primed in vivo with H-2 disparate spleen cells, it has not been previously possible to induce cytotoxic T lymphocyte (CTL) precursors (CTLp) in vitro by this type of challenge. In this work, we demonstrate that the inability to cross challenge in vitro is due to the existence of inhibitory effects that can be obviated by cell fractionation, and to insufficient priming in vivo. BALB/c CTLp (H-2d) that have been repeatedly primed in vivo with B10.D2 can be challenged in vitro with C57BL10/J (H-2b) or B10.BR (H-2k)-adherent cells to generate CTL able to lyse B10.D2 (H-2d) target cells. The H-2 restriction properties of the cross-challenged CTL specific for MIHA were analyzed by using the technique of cold target competition. Within the limits of detection in bulk cultures, the entire response appeared to be H-2 unrestricted, whether the cross challenge was with intact C57BL10/J-adherent cells, or with membrane fragments of C57BL10/J presented by BALB/c adherent cells. The frequency of CTLp responsive to cross challenge was analyzed by limiting dilution, with cold target competition at each cell number to establish the restriction properties of the MIHA-specific CTL induced. We were able to detect two subsets of H-2-unrestricted CTLp responsive to intact C57BL10/J-adherent cells; one present at high frequency (1/250 T cells) and subject to suppressive effects at high cell number, and a second present at lower frequency (1/9800 T cells). There appeared to be a relatively infrequent subset of H-2-restricted CTLp as well (1/52,500 T cells). The frequency of CTLp responsive to cross challenge is of comparable magnitude to the frequency of H-2-restricted CTLp responsive to H-2-matched cells bearing MIHA. These observations are discussed in relationship to immunodominance and clonal dominance effects in the response to MIHA.     

An analysis of B cell memory. I. Interaction of helper and suppressor effects in the in vitro expression of IgM memory.

The pfc response of Srbc primed IgM memory cells has been characterized by limiting dilution analysis in vitro, in which LPS was used to maximize the response of spleen cells to Srbc. The analysis suggested that, even under these conditions, expression of B cell memory was not directly assayed and cell collaboration effects were still basic to the system.Two types of cells, as defined by function, appeared necessary to elicit optimal clonal proliferation of IgM B memory cells: firstly, helper T cells were essential for B cell proliferation even with LPS present in culture. Under appropriate conditions, helper activity could be provided by normal thymus cells. Secondly, activated T cells were required for the maximal conversion of normal thymus cells to helper cells. A third activity, T cell-dependent suppression, was observed at high cell doses. The implications of these results and the need for a comprehensive analysis of in vitro conditions for each individual type of experiment is discussed.     

Leaf optical properties during and after drought stress in triticale and maize genotypes differing in drought tolerance

The effect of a short (7 days) and prolonged (14 days) soil drought (D) on leaf optical properties (R reflectance, T transmittance and A absorbance) in PAR and NIR range of irradiation, and on changes in leaf water potential (ψ), leaf injury index (LI), leaf thickness (LT) and chlorophyll (a + b) content (Chl) was studied for maize and triticale genotypes differing in drought tolerance. Under control conditions (C) leaves of maize in comparison to triticale were better hydrated, were thicker and had higher content of chlorophyll (a + b). In non-stressed plants, small differences were observed in measurements of R, T and A. In the range from 500 to 600 nm, the differences between D-resistant and D-sensitive were observed only in transmittance (T) and in range from 700 to 1,100 nm in absorbance (A). In genotypes belonging to the group of D-sensitive T in PAR range and A in NIR range were two times higher than in D-resistant ones. However, in NIR range R for D-sensitive genotypes was lower than for D-resistant ones. The drought stress caused the decrease in ψ, Chl, LT and the increase in leaf injury index (LI). Soil drought applied within 14 days caused larger changes in these physiological characters in comparison to 7 days drought. The observed harmful influence of drought was more visible for maize than triticale. Moreover for genotypes belonging to D-sensitive ones, changes were larger than for D-resistant ones. Similar to changes in ψ, LT and LI drought stress caused changes in leaf optical properties parameters R, A and T. In the PAR range, the highest changes were observed in R, whereas changes in T and A, which were not considerable. Both in maize and triticale, increase in R was higher in plants subjected to 14 days drought than in plants exposed to drought for 7 days. In maize, increase in R was larger for D-sensitive genotype. For both species, changes in T and A of PAR range were small. In NIR range, an increase in R and A, and decrease in T were observed. After 7 days of recovery in plants subjected to shorter period of drought significant differences were still visible in most cases. The same was observed for ψ, LT, LI and Chl parameters. It shows that the period of 7 days rehydration is too short to remove the injuries caused by drought stress. This results indicate that measurements of R, T and A might be useful in practical application for the estimation of the drought tolerance level. Some limitations in the practical application for plant breeding may be caused by relatively high cost of necessary equipment.     

A quantitative analysis of the interplay of environment,neighborhood, and cell state in 3D spheroids

Cells react to their microenvironment by integrating external stimuli into phenotypic decisions via an intracellular signaling network. To analyze the interplay of environment, local neighborhood, and internal cell state effects on phenotypic variability, we developed an experimental approach that enables multiplexed mass cytometric imaging analysis of up to 240 pooled spheroid microtissues. We quantified the contributions of environment, neighborhood, and intracellular state to marker variability in single cells of the spheroids. A linear model explained on average more than half of the variability of 34 markers across four cell lines and six growth conditions. The contributions of cell‐intrinsic and environmental factors to marker variability are hierarchically interdependent, a finding that we propose has general implications for systems‐level studies of single‐cell phenotypic variability. By the overexpression of 51 signaling protein constructs in subsets of cells, we also identified proteins that have cell‐intrinsic and cell‐extrinsic effects. Our study deconvolves factors influencing cellular phenotype in a 3D tissue and provides a scalable experimental system, analytical principles, and rich multiplexed imaging datasets for future studies.     

Chlorophyll distribution in the stems and trunk of beech trees

The distribution of chlorophyll was examined in cross-sections of 2- and 6-year-old stems as well as in the bark of the stump trunk of beech trees, utilising chlorophyll autofluorescence. The investigations were conducted using a confocal microscope. The tests carried out on 2 – 6-year old stems showed a large presence of chlorophyll in the bark, in primary and secondary rays as well as in the pith, but smaller amounts in wood parenchyma cells. There was no chlorophyll in the cork, sclerenchyma: in wood in vessels, tracheids and fibers. A reduction in the chlorophyll content in 6-year-old stems was not observed. In the bark of the trunk, chlorophyll was found in large amounts directly under the cork and in vestigial amounts in the primary phloem.     

The Systemic Cytokine Environment Is Permanently Altered in Multiple Myeloma

Multiple myeloma (MM) is an incurable bone marrow malignancy of the B cell lineage. Utilizing multiplex Luminex technology we measured levels of 25 cytokines in the plasma of normal donors (n = 177), those with monoclonal gammopathy of undetermined significance (n = 8), and MM patients (n = 55) with either active disease, on treatment, or in remission. The cytokine levels were compared between normal donors and MM patients as well as between various phases of MM, and discriminant analysis was used to create a predictive classification model based on the differentially expressed cytokines. Evaluating age- and gender-dependence of cytokine expression, we determined that with age there is a shift toward a pro-inflammatory environment. Moreover, we observed a strong gender bias in cytokine expression. However, the profile of differentially expressed cytokines was heavily skewed toward an anti-inflammatory, pro-tumorigenic response in patients with MM. Significantly, our predictive model placed all patients in remission in the same category as those with active disease. Thus, our study demonstrates that the homeostasis of systemic cytokines is not restored when MM patients enter remission, suggesting that once an individual has cancer, the microenvironment is permanently altered and the system is primed for a relapse.