Codelivery of the chemokine CCL3 by an adenovirus-based vaccine improves protection from retrovirus infection

Processing and presentation of vaccine antigens by professional antigen-presenting cells (APCs) is of great importance for the efficient induction of protective immunity. We analyzed whether the efficacy of an adenovirus-based retroviral vaccine can be enhanced by coadministration of adenovirus-encoded chemokines that attract and stimulate APCs. In the Friend retrovirus (FV) mouse model we coexpressed CCL3, CCL20, CCL21, or CXCL14 from adenoviral vectors, together with FV Gag and Env antigens, and then analyzed immune responses and protection from pathogenic FV infection. Although most tested chemokines did not improve protection against FV challenge, mice that received adenoviral vectors encoding CCL3 together with FV antigens showed significantly better control over viral loads and FV-induced disease than mice immunized with the viral antigens only. Improved protection correlated with enhanced virus-specific CD4+ T cell responses and higher neutralizing antibody titers. To apply these results to an HIV vaccine, mice were immunized with adenoviral vectors encoding the HIV antigens Env and Gag-Pol and coadministered vectors encoding CCL3. Again, this combination vaccine induced higher virus-specific antibody titers and CD4+ T cell responses than did the HIV antigens alone. These results indicate that coexpression of the chemokine CCL3 by adenovirus-based vectors may be a promising tool to improve antiretroviral vaccination strategies.     

Patterns of shoot architecture in locally adapted populations are linked to intraspecific differences in gene regulation

? Shoot architecture, including the number and location of branches, is a crucial aspect of plant function, morphological diversification, life history evolution and crop domestication. ? Genes controlling shoot architecture are well characterized in, and largely conserved across, model flowering plant species. The role of these genes in the evolution of morphological diversity in natural populations, however, has not been explored. ? We identify axillary meristem outgrowth as a primary driver of divergent branch number and life histories in two locally adapted populations of the monkeyflower, Mimulus guttatus. ? Furthermore, we show that MORE AXILLARY GROWTH (MAX) gene expression strongly correlates with natural variation in branch outgrowth in this species, linking modification of the MAX-dependent pathway to the evolutionary diversification of shoot architecture.     

Heparan Sulfate Biosynthesis Enzymes in Embryonic Stem Cell Biology

Embryonic stem (ES) cells are derived from the inner cell mass of the blastocyst and can give rise to all cell types in the body. The fate of ES cells depends on the signals they receive from their surrounding environment, which either promote self-renewal or initiate differentiation. Heparan sulfate proteoglycans are macromolecules found on the cell surface and in the extracellular matrix. Acting as low-affinity receptors on the cell surface, heparan sulfate (HS) side chains modulate the functions of numerous growth factors and morphogens, having wide impact on the extracellular information received by cells. ES cells lacking HS fail to differentiate but can be induced to do so by adding heparin. ES cells defective in various components of the HS biosynthesis machinery, thus expressing differently flawed HS, exhibit lineage-specific effects. Here we discuss recent studies on the biological functions of HS in ES cell developmental processes. Since ES cells have significant potential applications in tissue/cell engineering for cell replacement therapies, understanding the functional mechanisms of HS in manipulating ES cell growth in vitro is of utmost importance, if the stem cell regenerative medicine from scientific fiction ever will be made real.     

Cooperation of Non-Effective Concentration of Glutamatergic System Modulators and Antioxidant Against Oxidative Stress Induced by Quinolinic Acid

Excessive formation of reactive oxygen species (ROS) and disruption of glutamate uptake have been hypothesized as key mechanisms contributing to quinolinic acid (QA)-induced toxicity. Thus, here we investigate if the use of diphenyl diselenide (PhSe)(2), guanosine (GUO) and MK-801, alone or in combination, could protect rat brain slices from QA-induced toxicity. QA (1?mM) increased ROS formation, thiobarbituric acid reactive substances (TBARS) and decreased cell viability after 2?h of exposure. (PhSe)(2) (1?μM) protected against this ROS formation in the cortex and the striatum and also prevented decreases in cell viability induced by QA. (PhSe)(2) (5?μM) prevented ROS formation in the hippocampus. GUO (10 and 100?μM) blocked the increase in ROS formation caused by QA and MK-801 (20 and 100?μM) abolished the pro-oxidant effect of QA. When the noneffective concentrations were used in combination produced a decrease in ROS formation, mainly (PhSe)(2)?+?GUO and (PhSe)(2)?+?GUO?+?MK-801. These results demonstrate that this combination could be effective to avoid toxic effects caused by high concentrations of QA. Furthermore, the data obtained in the ROS formation and cellular viability assays suggest different pathways in amelioration of QA toxicity present in the neurodegenerative process.     

Prostaglandin receptors EP and FP are regulated by estradiol and progesterone in the uterus of ovariectomized rats

Background  

Prostaglandins are important for female reproduction. Prostaglandin-E2 acts via four different receptor subtypes, EP1, EP2, EP3 and EP4 whereas prostaglandin-F2alpha acts through FP. The functions of prostaglandins depend on the expression of their receptors in different uterine cell types. Our aim was to investigate the expression of EPs and FP in rat uterus and to identify the regulation by estradiol, progesterone and estrogen receptor (ER) selective agonists.     

NMR investigations of the dual targeting peptide of Thr-tRNA synthetase and its interaction with the mitochondrial Tom20 receptor in Arabidopsis thaliana

Most mitochondrial proteins are synthesized in the cytosol as precursor proteins containing an N-terminal targeting peptide and are imported into mitochondria through the import machineries, the translocase of the outer mitochondrial membrane (TOM) and the translocase of the inner mitochondrial membrane (TIM). The N-terminal targeting peptide of precursor proteins destined for the mitochondrial matrix is recognized by the Tom20 receptor and plays an important role in the import process. Protein import is usually organelle specific, but several plant proteins are dually targeted into mitochondria and chloroplasts using an ambiguous dual targeting peptide. We present NMR studies of the dual targeting peptide of Thr-tRNA synthetase and its interaction with Tom20 in Arabidopsis?thaliana. Our findings show that the targeting peptide is mostly unstructured in buffer, with a propensity to form α-helical structure in one region, S6-F27, and a very weak β-strand propensity for Q34-Q38. The α-helical structured region has an amphiphilic character and a φχχφφ motif, both of which have previously been shown to be important for mitochondrial import. Using NMR we have mapped out two regions in the peptide that are important for Tom20 recognition: one of them, F9-V28, overlaps with the amphiphilic region, and the other comprises residues L30-Q39. Our results show that the targeting peptide may interact with Tom20 in several ways. Furthermore, our results indicate a weak, dynamic interaction. The results provide for the first time molecular details on the interaction of the Tom20 receptor with a dual targeting peptide. DATABASE: The backbone chemical shift assignments for ThrRS-dTP(2-60) have been deposited with the Biological Magnetic Resonance Bank (BMRB) under the accession code 18248 STRUCTURED DIGITAL ABSTRACT: ThrRS-dTP?and?Tom20-4?bind?by?nuclear magnetic resonance?(View interaction).     

Effect of grazing on carbon stocks and assimilate partitioning in a Tibetan montane pasture revealed by 13CO2 pulse labeling

Since the late 1950s, governmental rangeland policies have changed the grazing management on the Tibetan Plateau (TP). Increasing grazing pressure and, since the 1980s, the privatization and fencing of pastures near villages has led to land degradation, whereas remote pastures have recovered from stronger overgrazing. To clarify the effect of moderate grazing on the carbon (C) cycle of the TP, we investigated differences in below‐ground C stocks and C allocation using in situ ¹³CO₂ pulse labeling of (i) a montane Kobresia winter pasture of yaks, with moderate grazing regime and (ii) a 7‐year‐old grazing exclosure plot, both in 3440 m asl. Twenty‐seven days after the labeling, ¹³C incorporated into shoots did not differ between the grazed (43% of recovered ¹³C) and ungrazed (38%) plots. In the grazed plots, however, less C was lost by shoot respiration (17% vs. 42%), and more was translocated below‐ground (40% vs. 20%). Within the below‐ground pools, <2% of ¹³C was incorporated into living root tissue of both land use types. In the grazed plots about twice the amount of ¹³C remained in soil (18%) and was mineralized to CO₂ (20%) as compared to the ungrazed plots (soil 10%; CO₂ 9%). Despite the higher contribution of root‐derived C to CO₂ efflux, total CO₂ efflux did not differ between the two land use types. C stocks in the soil layers 0–5 and 5–15 cm under grazed grassland were significantly larger than in the ungrazed grassland. However, C stocks below 15 cm were not affected after 7 years without grazing. We conclude that the larger below‐ground C allocation of plants, the larger amount of recently assimilated C remaining in the soil, and less soil organic matter‐derived CO₂ efflux create a positive effect of moderate grazing on soil C input and C sequestration.