Clonal growth in a species-rich grassland: Results of a 20-year fertilization experiment

We investigated the influence of fertilization on the abundance of species with different clonal growth characteristics using the data from a 20-year fertilization experiment from the Laelatu wooded meadow, in Estonia. The experiment comprised four different fertilization treatments and created a gradient of nitrogen availability. The vegetation composition was recorded every year by measuring the proportions of aboveground biomass for all species. For each species, four parameters of vegetative propagation were measured: speed of ramet vegetative mobility (annual increment of rhizome length), frequency of rhizome branching, placement of branches, and ramet life span. The weighted average of each parameter was calculated for each plot both at the beginning and at the end of the experiment using the relative abundances of the species in the plot as weights. The community changes resulting from the fertilization are reflected in the significant changes of the average values of all studied clonal growth parameters. Increased levels of phosphorus and potassium led to a community with an increased average vegetative mobility and rhizome branching. Both of these traits, however, declined with the increasing availability of nitrogen. The proportion of species with long-living ramets in the community decreased with the increase in the productivity irrespective of the fertilizer used. There was a strong positive correlation between the average ramet life span of the community and the number of species on the plot. We concluded that fertilization increased the ramet turnover rate in this meadow community and reduced species richness. Thus, our results contradict the prediction of a higher ramet turnover rate in species-rich compared to the species-poor grasslands.     

The dependence of Fluorescein-PE fluorescence intensity on lipid bilayer state. Evaluating the interaction between the probe and lipid molecules

The degree of dependence of a lipid bilayer's surface properties on its conformational state is still an unresolved question. Surface properties are functions of molecular organization in the complex interfacial region. In the past, they were frequently measured using fluorescence spectroscopy. Since a fluorescent probe provides information on its local environment, there is a need to estimate the effect caused by the probe itself. In this paper, we address this question by calculating how lipid head-group orientation effects the fluorescence intensity of Fluorescein-PE (a probe that is sensitive to surface potential). In the theoretical model assumed the lipid bilayer state and the interactions between the charged fluorescent probe and the surrounding lipid molecules was evaluated. The results of this theoretical analysis were compared with experimentally obtained data. A lipid bilayer formed from DPPC was chosen as the experimental system, since it exhibits all the major conformational states within a narrow temperature range of 30 degrees C-45 degrees C. Fluorescein-PE fluorescence intensity depends on local pH, which in turn is sensitive to local electrostatic potential in the probe's vicinity. This local electrostatic potential is generated by lipid head-group dipole orientation. We have shown that the effect of the probe on lipid bilayer properties is limited when the lipid bilayer is in the gel phase, whereas it is more pronounced when the membrane is liquid-crystalline. This implies that Fluorescein-PE is a good reporter of local electrostatic fields when the lipid bilayer is in the gel phase, and is a poor reporter when the membrane is in the liquid-crystalline state.     

Release of phospholipids from colloidal particles to polymeric surfaces

This study presents a small-scale polymerization of high molecular weight methyl methacrylate/n-butyl acrylate (MMA/n-BA) colloidal particles that are synthesized in an aqueous environment in the presence of phospholipid hydrogenated soybean phosphatidylcholine (HSPC) molecules that also serve as the particle stabilizing agents. When such particles coalesce to form polymeric films, they release phospholipids, which, in turn, form organized structures near the film-air (F-A) interface. Diffusion and mobility of phospholipid molecules are affected not only by their compatibility with colloidal particles but also by electrolyte environments of colloidal dispersions. When Na(+), K(+), and Ca(2+) counterions are added to MMA/n-BA aqueous colloidal dispersions stabilized with HSPC, and such films are coalesced, different degrees of diffusion of HSPC to the F-A interface exist, depending on the counterion, and conformational changes of HSPC result. For example, in the presence of Ca(2+), HSPC molecules collapse entropically to form random surface layers, as opposed to smaller Na(+) and K(+), which force amphiphilic HSPC ends to align preferentially parallel to the film surface. These studies show that it is possible to design stimuli-response colloidal systems triggered by chemical environments of active molecules on colloidal polymer particles.     

Increased activity and expression of matrix metalloproteinase-9 in a rat model of distal colitis

Matrix metalloproteinases may play a role in tissue remodelling and destruction associated with inflammation. We investigated activity and expression of matrix metalloproteinases in a rat model of colitis and tested the therapeutic potential of a synthetic inhibitor (CGS-27023-A). Colitis was induced by dextran sulphate sodium (at 5% in drinking water for 5 days) in a group of eight rats, whereas a matched control group received plain water. Activity and expression of matrix metalloproteinases were measured in colonic tissue homogenates using zymography and Western blot on days 3 and 5 after induction of colitis. In another set of experiments, two groups of colitic rats (20 per group) were treated with CGS-27023-A (20 mg/kg) or vehicle, respectively. On days 5 and 14, colonic mucosal lesions were blindly scored by microscopic examination. Induction of colitis led to a significant upregulation of matrix metalloproteinase-9 protein and its activity, but no change in matrix metalloproteinase-2 activity was observed. Treatment with CGS-27023-A significantly decreased the extent and severity of epithelial injury but did not influence mucosal repair. We conclude that increased activity of matrix metalloproteinases may contribute to epithelial damage in this model of colitis.     

Distinct mechanisms underlie sperm-induced and protease-induced oolemma block to sperm penetration

Fertilization of a mouse egg results in modification of the cytoplasmatic membrane (oolemma) which makes fusion with additional sperm impossible. CD9 is a transmembrane protein reported to be responsible for gamete fusion. Since the molecular mechanism of zygote membrane modification after fertilization remains unknown, we were interested to check whether lack of CD9 is the reason for non-penetrability of zona-free zygotes. We wanted also to determine the effect of different methods of zona pellucida removal on the presence of CD9 on the surface of unfertilized eggs and their ability to be fertilized afterwards. We demonstrated that CD9 is present on the surface of both zygotes and parthenogenotes. We showed also that the treatment of eggs with pronase completely removes CD9 from the membrane of eggs making them infertile. Eggs treated with chymotrypsin and acid Tyrode still posses CD9 on their surface and remain fertile. The results of our experiments indicate that modification of the zygote oolemma does not involve a lack of CD9. We cannot exclude however, that the amount of CD9 decreases after fertilization. In addition, our studies indicate that the previously reported infertility of eggs treated with different proteases may result from the decrease or removal of CD9 and probably other proteins responsible for gamete fusion from the surface of eggs.     

Prickle and Strabismus form a functional complex to generate a correct axis during planar cell polarity signaling

Frizzled (Fz) signaling regulates the establishment of planar cell polarity (PCP). The PCP genes prickle (pk) and strabismus (stbm) are thought to antagonize Fz signaling. We show that they act in the same cell, R4, adjacent to that in which the Fz/PCP pathway is required in the Drosophila eye. We demonstrate that Stbm and Pk interact physically and that Stbm recruits Pk to the cell membrane. Through this interaction, Pk affects Stbm membrane localization and can cause clustering of Stbm. Pk is also known to interact with Dsh and is thought to antagonize Dsh by affecting its membrane localization. Thus our data suggest that the Stbm/Pk complex modulates Fz/Dsh activity, resulting in a symmetry-breaking step during polarity signaling.     

Photosynthetic UV-B Response of Beech (Fagus sylvatica L.) Saplings

Cloned saplings of beech (7-y-old) were exposed to enhanced UV-B irradiation (+25 %) continuously over three growing seasons (1999–2001). Analysis of CO₂ assimilation, variable chlorophyll (Chl) a fluorescence, and pigment composition was performed in late summer of the third growing season to evaluate the influence of long-term elevated UV-B irradiation. This influence was responsible for the stimulation of the net assimilation rate (P _N) over a range of irradiances. The increase in P _N was partially connected to increase of the area leaf mass, and thus to the increased leaf thickness. Even a higher degree of UV-B induced stimulation was observed at the level of photosystem 2 (PS2) photochemistry as judged from the irradiance response of electron transport rate and photochemical quenching of Chl a. The remarkably low irradiance-induced non-photochemical quenching of maximum Chl a fluorescence (NPQ) in the UV-B plants over the entire range of applied irradiances was attributed both to the reduced demand on non-radiative dissipation processes and to the considerably reduced contribution of the quenching localised in the inactivated PS2 reaction centres. Neither the content of Chls and total carotenoids expressed per leaf area nor the contents of lutein, neoxanthin, and the pool of xanthophyll cycle pigments (VAZ) were affected under the elevated UV-B. However, the contributions of antheraxanthin (A) and zeaxanthin (Z) to the entire VAZ pool in the dark-adapted UV-B treated plants were 1.61 and 2.14 times higher than in control leaves. Surprisingly, the retained A+Z in UV-B treated plants was not accompanied with long-term down-regulation of the PS2 photochemical efficiency, but it facilitated the non-radiative dissipation of excitation energy within light-harvesting complexes (LHC) of PS2. Thus, in the beech leaves the accumulation of A+Z, induced by other factors than excess irradiance itself, supports the resistance of PS2 against combined effects of high irradiance and elevated UV-B.     

Evaluation and comparison of random amplification of polymorphic DNA, pulsed-field gel electrophoresis and ADSRRS-fingerprinting for typing Serratia marcescens outbreaks

Amplification of DNA fragments surrounding rare restriction sites (ADSRRS-fingerprinting) is a novel assay based on suppression of polymerase chain reaction (PCR). This phenomenon allows the amplification of only a limited subset of DNA fragments, since only those with two different oligonucleotides ligated at the ends of complementary DNA strands are amplified in the PCR. The DNA fragments can be easily analyzed on polyacrylamide gels, stained with ethidium bromide. We have implemented this method using a set of clinical Serratia marcescens isolates from three outbreaks ongoing in the Public Hospital in Gdańsk (Poland). Clustering of ADSRRS-fingerprinting data matched epidemiological, microbiological, random amplification of polymorphic DNA (RAPD) and pulsed-field gel electrophoresis (PFGE) data. Based on this study, we found that there is at least a similar power of discrimination between the present 'gold-standard' PFGE and the novel method, ADSRRS-fingerprinting. Although the ADSRRS-fingerprinting method may appear to be more complex than the RAPD technique, we found it fast and reproducible.