Benzyladenine effects on the development of the photosynthetic apparatus in Zea mays: Studies on photosynthetic activity, enzymes and (etio)chloroplast ultrastructure

Primary leaf segments from 8-day-old dark-grown, and from 4- and 8-day-old light-grown seedlings of Zea mays L. cv. Fronica, were treated with 10^-bM benzyladenine (BA) in the dark for 14 h. The segments were then studied after an exposure to light for 14 h. Photosynthetic activity (O₂ evolution and CO₂ fixation) and chlorophyll accumulation were stimulated by BA in dark-grown leaf segments with etioplastids in the earliest stage of development. In these segments BA stimulated the activities of ribulose-1,5-bisphosphate carboxylase (EC 4.1.1.39), phosphoenolpyruvate carboxylase (EC 4.1.1.31), NADP⁺-malic enzyme (EC 1.1.1.40) and pyruvate, orthophosphate dikinase (EC 2.7.9.1). In segments taken from 4- and 8-day light-grown seedlings, BA did not enhance the photosynthetic activity nor the chlorophyll accumulation. The activity of the enzymes mentioned above, was significantly enhanced by the BA-treatment. BA mainly affected grana stacking in mesophyll cell chloroplasts in primary leaf segments taken from 3- to 5-day light-grown seedlings. Stroma thylakoid development was stimulated only in leaf segments from 3-day-old plants. At the same time BA accelerated grana loss in chloroplasts of bundle sheath cells, a typical phenomenon of development in such chloroplasts. Stroma thylakoid length in these chloroplasts increased by a BA treatment in segments from 3- and 4-day light-grown plants. A significantly higher number of chloroplasts was only observed with segments taken from 8-day light-grown seedlings and treated with BA. The etiochloroplast number in segments taken from 8-day etiolated plants was significantly higher in BA-treated segments after 26 h illumination. In etiochloroplasts from both mesophyll and bundle sheath cells, BA enhanced grana stacking after illumination for 4 h or more, whereas stroma membrane length was significantly higher only after 26 h light. It is concluded that the effects of BA depend on the developmental stage. BA accelerates the development of mesophyll and bundle sheath cell (etio)chloroplasts, but does not affect the ultrastructure of mature chloroplasts.     

Some aspects of the synthesis of long-lived messenger ribonucleoproteins in developing rye embryos

Untranslated messenger ribonucleoproteins (mRNPs informosomes) are present in developing rye (Secale cereale, L. cv. Celestijner) embryos throughout the last 5 weeks of seed formation. Ribosomal as well as non-ribosomal ribonucleoproteins are formed continuously both in primary axes and scutella until any synthesis of macromolecules stops upon dessication. The content of preformed messengers in the primary axes of precociously harvested rye grains increases as a function of embryo development. This increase of the template load of the primary axes results from a continuous accumulation of qualitatively identical mRNPs. Germination experiments demonstrated that at least most of the preformed messengers are not required for the germination process. Their function is discussed in terms of selective adaptation to unfavorable conditions.Abbreviations SHB standard HEPES buffer - PB polysome buffer - HEPES N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid - SDS sodium dodecyl sulphate - mRNP messenger ribonucleoprotein - TMV-RNA tobacco mosaic virus RNA - PM postmitochondrial     

Preformed and newly synthesized messenger RNA in germinating wheat embryos

In 6 h germinated wheat (Triticum aestivum L. cv. Cama) embryos, more than half of the messenger RNAs are actively involved in translation. Neither preformed nor newly synthesized poly A⁺-RNA is translated preferentially. Germination in the presence of cordycepin showed that the half-life of the templates is about 2 h and that the newly synthesized messengers are essential to support protein synthesis in the embryo from the first hours of germination. Most of the messenger RNAs in 6 h germinated embryos are newly synthesized. The polypeptides coded for by either the endogenous messenger ribonucleoproteins or purified poly A⁺-RNA from both dry and germinated embryos are qualitatively identical; minor quantitative differences can however be observed.Abbreviations HEPES N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid - EDTA ethylenediaminetetraacetic acid - SDS sodium dodecyl sulfate - TCA trichloroacetic acid - mRNP messenger ribonucleoprotein - poly A⁺-RNA polyadenylic acid containing RNA - PB polysome buffer - GM germination medium     

Database on the structure of small ribosomal subunit RNA.

About 8600 complete or nearly complete sequences are now available from the Antwerp database on small ribosomal subunit RNA. All these sequences are aligned with one another on the basis of the adopted secondary structure model, which is corroborated by the observation of compensating substitutions in the alignment. Literature references, accession numbers and detailed taxonomic information are also compiled. The database can be consulted via the World Wide Web at URL http://rrna.uia.ac.be/ssu/     

Database on the structure of large ribosomal subunit RNA.

The rRNA WWW Server at URL http://rrna.uia.ac.be/ now provides a database of 496 large subunit ribosomal RNA sequences. All these sequences are aligned, incorporate secondary structure information, and can be obtained in a number of formats. Other information about the sequences, such as literature references, accession numbers and taxonomic information is also available and searchable. If necessary, the data on the server can also be obtained by anonymous ftp.     

Establishment of a Murine Graft-versus-Myeloma Model Using Allogeneic Stem Cell Transplantation

BackgroundMultiple myeloma (MM) is a malignant plasma cell disorder with poor long-term survival and high recurrence rates. Despite evidence of graft-versus-myeloma (GvM) effects, the use of allogeneic hematopoietic stem cell transplantation (allo-SCT) remains controversial in MM. In the current study, we investigated the anti-myeloma effects of allo-SCT from B10.D2 mice into MHC-matched myeloma-bearing Balb/cJ mice, with concomitant development of chronic graft-versus-host disease (GvHD).ConclusionsWe successfully established an immunocompetent murine model of graft-versus-myeloma. This is the first murine GvM model using immunocompetent mice that develop MM which closely resembles human MM disease and that are treated after disease establishment with an allo-SCT. Importantly, using TCR Vβ spectratyping, we also demonstrated the presence of GvM unique responses potentially associated with the curative capacity of this immunotherapeutic approach.     

An experimental test of the role of environmental temperature variability on ectotherm molecular, physiological and life-history traits: implications for global warming

Global climate change is one of the greatest threats to biodiversity; one of the most important effects is the increase in the mean earth surface temperature. However, another but poorly studied main characteristic of global change appears to be an increase in temperature variability. Most of the current analyses of global change have focused on mean values, paying less attention to the role of the fluctuations of environmental variables. We experimentally tested the effects of environmental temperature variability on characteristics associated to the fitness (body mass balance, growth rate, and survival), metabolic rate (VCO(2)) and molecular traits (heat shock protein expression, Hsp70), in an ectotherm, the terrestrial woodlouse Porcellio laevis. Our general hypotheses are that higher values of thermal amplitude may directly affect life-history traits, increasing metabolic cost and stress responses. At first, results supported our hypotheses showing a diversity of responses among characters to the experimental thermal treatments. We emphasize that knowledge about the cellular and physiological mechanisms by which animals cope with environmental changes is essential to understand the impact of mean climatic change and variability. Also, we consider that the studies that only incorporate only mean temperatures to predict the life-history, ecological and evolutionary impact of global temperature changes present important problems to predict the diversity of responses of the organism. This is because the analysis ignores the complexity and details of the molecular and physiological processes by which animals cope with environmental variability, as well as the life-history and demographic consequences of such variability.     

A Novel Mouse Model for Multiple Myeloma (MOPC315.BM) That Allows Noninvasive Spatiotemporal Detection of Osteolytic Disease

Multiple myeloma (MM) is a lethal human cancer characterized by a clonal expansion of malignant plasma cells in bone marrow. Mouse models of human MM are technically challenging and do not always recapitulate human disease. Therefore, new mouse models for MM are needed. Mineral-oil induced plasmacytomas (MOPC) develop in the peritoneal cavity of oil-injected BALB/c mice. However, MOPC typically grow extramedullary and are considered poor models of human MM. Here we describe an in vivo-selected MOPC315 variant, called MOPC315.BM, which can be maintained in vitro. When injected i.v. into BALB/c mice, MOPC315.BM cells exhibit tropism for bone marrow. As few as 10⁴ MOPC315.BM cells injected i.v. induced paraplegia, a sign of spinal cord compression, in all mice within 3–4 weeks. MOPC315.BM cells were stably transfected with either firefly luciferase (MOPC315.BM.Luc) or DsRed (MOPC315.BM.DsRed) for studies using noninvasive imaging. MOPC315.BM.Luc cells were detected in the tibiofemoral region already 1 hour after i.v. injection. Bone foci developed progressively, and as of day 5, MM cells were detected in multiple sites in the axial skeleton. Additionally, the spleen (a hematopoietic organ in the mouse) was invariably affected. Luminescent signals correlated with serum myeloma protein concentration, allowing for easy tracking of tumor load with noninvasive imaging. Affected mice developed osteolytic lesions. The MOPC315.BM model employs a common strain of immunocompetent mice (BALB/c) and replicates many characteristics of human MM. The model should be suitable for studies of bone marrow tropism, development of osteolytic lesions, drug testing, and immunotherapy in MM.     

Database on the structure of large subunit ribosomal RNA.

The Antwerp database on large subunit ribosomal RNA now contains 607 complete or nearly complete aligned sequences. The alignment incorporates secondary structure information for each sequence. Other information about the sequences, such as literature references, accession numbers and taxonomic information is also available. Information from the database can be downloaded or searched on the rRNA WWW Server at URL http://rrna.uia.ac.be/