Enzymatic evidence for the key role of arginine in nitrogen translocation by arbuscular mycorrhizal fungi

Key enzymes of the urea cycle and (15)N-labeling patterns of arginine (Arg) were measured to elucidate the involvement of Arg in nitrogen translocation by arbuscular mycorrhizal (AM) fungi. Mycorrhiza was established between transformed carrot (Daucus carota) roots and Glomus intraradices in two-compartment petri dishes and three ammonium levels were supplied to the compartment containing the extraradical mycelium (ERM), but no roots. Time courses of specific enzyme activity were obtained for glutamine synthetase, argininosuccinate synthetase, arginase, and urease in the ERM and AM roots. (15)NH(4)(+) was used to follow the dynamics of nitrogen incorporation into and turnover of Arg. Both the absence of external nitrogen and the presence of L-norvaline, an inhibitor of Arg synthesis, prevented the synthesis of Arg in the ERM and resulted in decreased activity of arginase and urease in the AM root. The catabolic activity of the urea cycle in the roots therefore depends on Arg translocation from the ERM. (15)N labeling of Arg in the ERM was very fast and analysis of its time course and isotopomer pattern allowed estimation of the translocation rate of Arg along the mycelium as 0.13 microg Arg mg(-1) fresh weight h(-1). The results highlight the synchronization of the spatially separated reactions involved in the anabolic and catabolic arms of the urea cycle. This synchronization is a prerequisite for Arg to be a key component in nitrogen translocation in the AM mycelium.     

Changes in Pelagic Bacteria Communities Due to Leaf Litter Addition

In many limnetic systems, the input of allochthonous organic matter, e.g., leaf litter, is a substantial source of dissolved organic carbon (DOC) for pelagic bacteria, especially in fall and winter when autochthonous DOC production is low. However, relatively little is known about community changes of pelagic lake bacteria due to leaf litter input which includes both the release of leaf leachates and microorganisms from the leaf litter into the surrounding water. Therefore, we have experimentally studied the effects of different types of leaf litter (Betula pendula, Fagus silvatica, and Pinus silvestris) on the pelagic bacterial community composition by adding leaves to different treatments of epilimnic water samples (unfiltered, 0.2 μm and 5.0 μm-pre-filtered) from humic Lake Grosse Fuchskuhle (Northeastern Germany). The addition of leaf litter led to a significant increase in DOC concentration in lake water, and each leaf litter type produced significantly different amounts of DOC (p = <0.001) as well as of specific DOC fractions (p = <0.001), except of polysaccharides. DGGE banding patterns varied over time, between types of leaf litter, and among treatments. Bacteria belonging to known bacterial phylotypes in the southwest basin of Lake Grosse Fuchskuhle were frequently found and even persisted after leaf litter additions. Upon leaf litter addition, α-proteobacteria (Azospirillum, Novosphingobium, and Sphingopyxis) as well as β-proteobacteria (Curvibacter and Polynucleobacter) were enriched. Our results indicate that supply of leaf litter DOM shifted the bacterial community in the surrounding water towards specific phylotypes including species capable of assimilating the more recalcitrant DOC pools. Statistical analyses, however, show that DGGE banding patterns are not only affected by DOC pools but also by treatment. This indicates that biological factors such as source community and grazing may be also important for shifts in bacterial community structure following leaf litter input into different lakes.     

Stimulation of AIDS lymphocytes with calcium ionophore (A23187) and phorbol ester (PMA): studies of cytoplasmic free Ca, IL-2 receptor expression, IL-2 production, and proliferation

We have studied whether the decreased lymphocyte proliferative responses of AIDS lymphocytes to stimulation by mitogens and antigens may be overcome when challenged with a combination of calcium ionophore A23187 and phorbol ester PMA. Comparison of the proliferative response of lymphocytes from nine patients with AIDS with the response of lymphocytes from nine control subjects showed that the response of AIDS lymphocytes was severely decreased when stimulated with PHA and no further response could be achieved by stimulation with A23187/PMA. On the other hand, no significant difference between the PHA-induced rise of cytoplasmic free calcium concentration ([Ca2+]1) in normal and AIDS lymphocytes was observed. The percentage of cells expressing IL-2 receptors (CD25) was also normal both after addition of PHA and after addition of A23187/PMA and the expression was normal on both CD4 and CD8 cells. The production of IL-2 in normal lymphocytes stimulated with A23187/PMA was 33 times higher than that after stimulation with PHA. In AIDS lymphocytes the production of IL-2 induced by all activators was severely decreased compared to control subjects, although the production of IL-2 after stimulation with A23187/PMA was higher than that in control lymphocytes after stimulation with PHA. The present study shows that a direct activation of protein kinase C combined with mobilization of cytoplasmic calcium does not overcome the lymphocyte proliferative deficiency of AIDS lymphocytes.     

DNA damage-dependent acetylation and ubiquitination of H2AX enhances chromatin dynamics

Chromatin reorganization plays an important role in DNA repair, apoptosis, and cell cycle checkpoints. Among proteins involved in chromatin reorganization, TIP60 histone acetyltransferase has been shown to play a role in DNA repair and apoptosis. However, how TIP60 regulates chromatin reorganization in the response of human cells to DNA damage is largely unknown. Here, we show that ionizing irradiation induces TIP60 acetylation of histone H2AX, a variant form of H2A known to be phosphorylated following DNA damage. Furthermore, TIP60 regulates the ubiquitination of H2AX via the ubiquitin-conjugating enzyme UBC13, which is induced by DNA damage. This ubiquitination of H2AX requires its prior acetylation. We also demonstrate that acetylation-dependent ubiquitination by the TIP60-UBC13 complex leads to the release of H2AX from damaged chromatin. We conclude that the sequential acetylation and ubiquitination of H2AX by TIP60-UBC13 promote enhanced histone dynamics, which in turn stimulate a DNA damage response.     

Reductive Dechlorination of Trichloroethylene (TCE) in Competition with Fe and Mn Oxides—Observed Dynamics in H2-dependent Terminal Electron Accepting Processes

The determination of hydrogen (H₂) concentration together with the products of microbial reduction reactions in a trichloroethylene dechlorinating system is conducted to delineate the ongoing predominant terminal electron accepting processes (TEAP). Formate was used as electron donor and synthetic Fe minerals or environmental samples were used as the substrate. Iron(III) and Mn(IV) reduction limited microbial dechlorination by the mixed anaerobic culture by decreasing the level of H₂ in the system. The H₂ measurements indicated that the H₂ concentration at which different TEAPs occur can overlap and thus these TEAPs can therefore occur concurrently rather than exclusively. Difference in Fe(III) bioavailability and hence, Fe(III) reduction partially explain this wide range. The distinction between dechlorination and other microbial reduction processes based on H₂ threshold values is not feasible under such conditions, though there appears to be a relation between the rates of H₂ consuming process and the observed H₂ level.     

Underground resource allocation between individual networks of mycorrhizal fungi

Fusions between individual mycelia of arbuscular mycorrhizal (AM) fungi have been observed in two-dimensional systems but never in soil systems. Here, phosphorus ((32)P) labelling was used to demonstrate nutrient transfer between individual mycelia and to investigate the possible role of anastomosis. Trifolium subterraneum colonized by Glomus mosseae were grown in root-retaining mesh bags, which were placed 20 cm apart. The mycelium of one plant, the donor, had access to (32)P-labelled soil placed adjacent to the mesh bag. Transfer of (32)P from the donor mycelium to the receiver plant was measured at three harvests. In a second-harvest control treatment the receiver was colonized by Glomus caledonium in order to determine whether transfer occurred by other means than hyphal fusions. Significant amounts of P were transferred to the receiver plant at the last harvests when the two mycelia of G. mosseae overlapped. The transfer probably occurred via anastomoses between the mycelia as no transfer of (32)P was detected between the mycelia of different fungi at the second harvest. The indicated ability of AM fungal mycelia to anastomose in soil has implications for the formation of large plant-interlinking functional networks, long-distance nutrient transport and retention of nutrients in readily plant-available pools.     

Evolution of plant RNA polymerase IV/V genes: evidence of subneofunctionalization of duplicated <Emphasis Type=Italic>NRPD2/NRPE2</Emphasis>-like paralogs in <Emphasis Type=Italic>Viola</Emphasis> (Violaceae)

Background  

DNA-dependent RNA polymerase IV and V (Pol IV and V) are multi-subunit enzymes occurring in plants. The origin of Pol V, specific to angiosperms, from Pol IV, which is present in all land plants, is linked to the duplication of the gene encoding the largest subunit and the subsequent subneofunctionalization of the two paralogs (NRPD1 and NRPE1). Additional duplication of the second-largest subunit, NRPD2/NRPE2, has happened independently in at least some eudicot lineages, but its paralogs are often subject to concerted evolution and gene death and little is known about their evolution nor their affinity with Pol IV and Pol V.     

Genome sequence of the naturally plasmid-free Lactobacillus plantarum strain NC8 (CCUG 61730)

Lactobacillus plantarum is a highly versatile lactic acid bacterium found in various ecological niches, such as fermented vegetable, meat, and dairy products and the gastrointestinal tract. We sequenced the genome of L. plantarum NC8, a naturally plasmid-free strain, which has been used as a model strain in many laboratories worldwide.     

Tooth and bone development in a Danish medieval mandible with unilateral absence of the mandibular canal.

A Danish anthropological collection of medieval human skeletons excavated in 1986 involves a mandible (No. 212) from an adult female born without the lower alveolar nerve and mandibular canal. It is believed that the defect has resulted in lack of tooth development on the affected side and that the mylohyoid nerve has partially compensated for this defect by development of teeth in localized areas. The defective mandibular dentition has caused a compensatory development of the alveolar process in the maxilla. The missing occlusal support has altered muscular traction on the mandible. This has caused an alteration in mandibular shape. Whether the asymmetric development of the mandible is caused by muscular dysfunction, by failure in angular growth apposition, or by a combination of these factors is discussed. The case presents valuable data in the ongoing discussion about the interaction between nerve tissue and tooth formation and about the interaction between occlusion, jaw morphology, and muscular traction. The study shows how archeological material in an interdisciplinary cooperation between archeological, embryological and orthodontic research can contribute to the clarification of current biological problems.     

Commensal Leucothoidae (Crustacea, Amphipoda) of the Ryukyu Archipelago, Japan. Part II: sponge-dwellers

Commensal leucothoid amphipods have been collected from the canals of their sponge hosts throughout the Ryukyu Archipelago, Japan. Eleven new species are described in the genus Leucothoe with valuable location data and host records. An identification key to sponge-dwelling Leucothoidae of the Ryukyu Archipelago is provided.