The importance of nitrogen-fixation for an invader of a coastal California grassland

Whether a novel trait of an invader directly contributes to increased establishment of that invader is a relatively unstudied question in plant ecology. Nitrogen (N)-fixing shrubs comprise a significant subset of grassland invaders worldwide, which suggests the potential importance of the novel trait of N-fixation in the invasion process. We indirectly tested the importance of N-fixation in the invasion of Genista monspessulana (French broom) in a California grassland by alleviating N and phosphorus (P) limitation to the grassland matrix. Grassland productivity was co-limited by N and P; N alone did not release the resident vegetation, and did not affect Genista performance. Genista was strongly limited by P: seedlings had more nodules, greater leaf N concentration, and higher growth and survival with P additions. When N was added with P, however, growth of the resident vegetation was 50?C70% greater than with N or P alone, accompanied by decreases in Genista performance. This suggests that the advantage conferred to Genista by N-fixation was dampened when the resident vegetation was released from nutrient limitation.     

Carbon allocation in ectomycorrhizal plants at limited and optimal N supply: an attempt at unraveling conflicting theories

With regard to mycorrhiza, conflicting theories try to explain how the balance between fungal demand for carbohydrates and the plant’s needs for nutrients varies, resulting in conflicting predictions. In order to evaluate current concepts, we investigated some metabolic parameters, which are indicative for plant carbon allocation in response to mycorrhization at limited and optimal N supply. Pinus pinaster seedlings were inoculated with living or dead (control) cultures of Pisolithus tinctorius, supplied with ammonium at 4 (limiting) or 7% d⁻¹ (non-limiting) N relative addition rate (RAR_N), and followed development for 29 days. Mycorrhizal colonization of roots was quantified by the determination of ergosterol. A series of enzymes (sucrose and trehalose metabolism, anaplerosis) and metabolites (soluble carbohydrate, including trehalose; fructose 2,6 bisphosphate, free amino acids) relevant in the C/N exchange between symbionts, and in the carbon allocation and sink strength within the plant were assayed for 2-day-intervals for up to 14 days, and at 5-day-intervals for the rest of the experiment. The first 10 days reflected the establishment of mycorrhizal interaction, and the carbon allocation to the root was higher in M plants independent of N supply. Following this period, carbon allocation became N-related, higher at low, and lower at high N supply. The belowground C investment of M plants was dependent on N availability, but not on N gain. Finally, increased belowground C allocation was accompanied by a shift from plant to fungal metabolism.     

The use of microsatellites to analyze relationships and to decipher homonyms and synonyms in Azorean apples (Malus?×?domestica Borkh.)

The objective of this study was to exploit the molecular and morphological variability present in Malus domestica to clarify the confused characterization of apple plantations in the Azores. Most Azorean apples are grown in orchards. They are usually given a local name, and sometimes the same name is used for different cultivars and varieties which share morphology and should be known by different names. Two-hundred samples of apples cultivated in the Azores were analyzed by use of ten microsatellites. The total number of alleles per locus ranged from 10 to 24, with a mean of 15.2. Heterozygosity was high, reflecting the high variability of the samples. Expected heterozygosity varied within a narrow range, from 0.74 to 0.88, whereas observed heterozygosity ranged from 0.41 to 0.98. The high genetic variability contributed to the high discriminating power, which ranged between 0.84 and 0.93. These microsatellites were used to unambiguously discriminate most of the tested apple cultivars on the basis of their allelic profiles. The rooted UPGMA tree organized most of the samples into thirteen main clusters, often with high bootstrap values. We identified 60 synonyms and 32 homonyms among the samples. Moreover, it was possible to relate each individual to its originating population and detect likely parent–offspring groups.     

Performance and Mortality of Rumex obtusifolius and R. crispus in Managed Grasslands are Affected by Nutrient Availability

The reasons for the greater incidence of Rumex obtusifolius in grasslands compared with R. crispus have never been investigated. In a small-plot field experiment in which seedlings of R. obtusifolius and R. crispus were transplanted into a sward dominated by Dactylis glomerata, the growth and survival of seedlings were monitored over three years under control and P, N, NP and NPK fertilizer treatments. The highest plants of R. obtusifolius and R. crispus were generally recorded in the N, NP and NPK treatments and the lowest in the control and P treatment. The maximum recorded heights of R. obtusifolius were 100?cm and of R. crispus 80?cm. The number of leaves per plant, weight of individual plants, cover and fertility were generally higher for R. obtusifolius than for R. crispus. 30% to 80% of transplanted R. obtusifolius plants flowered in the first (seeding) year and this contrasted with no flowering of transplanted R. crispus plants. After cutting, substantially better regrowth of R. obtusifolius was recorded compared to that of R. crispus and surviving R. crispus plants tended to show a gradual reduction in leaf number. The mortality of R. obtusifolius over three years ranged from 0% in the NPK treatment to 13% in the control, with an average of 8% over all treatments. The mortality of R. crispus ranged from 19% in the control to 94% in the NPK treatment with an average of 64% over all treatments. We concluded that R. obtusifolius is better adapted to growing in highly productive temperate grasslands than R. crispus because of its perennial character, larger plant size and higher fertility.     

Phylogenetic diversity of bacterial and archaeal communities inhabiting the saline Lake Red located in Sovata, Romania

Lake Red is one of the saline lakes which were formed as a consequence of salt massif dissolution at the foot of the Gurghiu Mountains (Central Romania) at the end of the nineteenth century. The lake water had approximately 15?% w/v salt content. Phylogenetic diversity of prokaryotes inhabiting the water and sediment of the lake was studied using cultivation and cultivation-independent methods following a sampling in spring 2009. According to the results of 16S rRNA gene-based denaturing gradient gel electrophoresis (DGGE), the richness of Bacteria was higher than Archaea on the basis of the number and position of dominant bands in the gel. Sequences from DGGE bands were affiliated with Gammaproteobacteria (Halomonas and Alkalilimnicola) and Bacteroidetes (Psychroflexus) as well as Euryarchaeota. Cultivation from five different saline media resulted in 101 bacterial strains of which Gammaproteobacteria (Halomonas, Marinobacter and Salinivibrio) were the most abundant. Firmicutes (Bacillus) and Alphaproteobacteria (Aurantimonas and Roseovarius) were also identified among the isolated strains. The 16S rRNA genes from 82 bacterial and 95 archaeal clones were also phylogenetically analyzed. Bacterial clones were related to various genera of Gammaproteobacteria (Alkalilimnicola, Alkalispirillum, Arhodomonas, Halomonas, Saccharospirillum), Bacteroidetes (Gracilimonas, Psychroflexus) and Alphaproteobacteria (Oceanicola, Roseinatronobacter, Roseovarius). All of the archaeal clones sequenced corresponded to a homologous cluster affiliated with Halopelagius.     

Identification of miRNAs Expression Profile in Gastric Cancer Using Self-Organizing Maps (SOM)

In this paper, an unsupervised artificial neural network was implemented to identify the patters of specific signatures. The network was based on the differential expression of miRNAs (under or over expression) found in healthy or cancerous gastric tissues. Among the tissues analyzes, the neural network evaluated 514 miRNAs of gastric tissue that exhibited significant differential expression. The result suggested a specific expression signature nine miRNAs (hsa-mir-21, hsa-mir-29a, hsa-mir-29c, hsa-mir-148a, hsa-mir-141, hsa-let-7b, hsa-mir-31, hsa-mir-451, and hsa-mir-192), all with significant values (p-value < 0.01 and fold change > 5) that clustered the samples into two groups: healthy tissue and gastric cancer tissue. The results obtained “in silico” must be validated in a molecular biology laboratory; if confirmed, this method may be used in the future as a risk marker for gastric cancer development.     

Hybrid aspens responses to alkalisation of soil: growth, leaf structure, photosynthetic rate and carbohydrates

Interactions between photosynthetic rate, transpiration, content of soluble carbohydrates, leaf dimensions and structural parameters and growth of hybrid aspen (Populus tremula?×?P. tremuloides Michx.) were studied in relation to soil pH and chemical composition. The investigations were conducted in two plantations on former agricultural lands in North Estonia. One plantation was established on a territory influenced for a long time by alkaline cement dust pollution before plantation establishment and the other on an unpolluted territory. At pH 7.4 and higher concentrations of Ca, K, Mg, N and P in soil on the polluted area inhibition of height growth and diameter at breast height, leaf area and dry mass was observed compared to the unpolluted plantation with an optimum soil pH of 6.7. Differences in hybrid aspen leaves in the two plantations were related to the lower net photosynthetic and transpiration rates and higher starch and sucrose contents in the polluted plantation. Leaves from alkaline soil had a thicker palisade mesophyll layer and lower number of stomata at abaxial epidermis. The relatively low N concentration in leaves in the polluted area was associated with the low height and diameter at breast height of trees.     

Effects of β-Cyclodextrin on the Structure of Sphingomyelin/Cholesterol Model Membranes

The interaction of β-cyclodextrin (β-CD) with mixed bilayers composed of sphingomylein and cholesterol (Chol) above and below the accepted stable complexation ratio (67:33) was investigated. Membranes with the same (symmetric) and different (asymmetric) compositions in their inner and outer leaflets were deposited at surface pressures of 20, 30, and 40 mN/m at the solid-liquid interface. Using neutron reflectometry, membranes of various global molar ratios (defined as the sum of the molar ratios of the inner and outer leaflets), were characterized before and after β-CD was added to the subphase. The structure of bilayers with global molar ratios at or above the stable complexation ratio was unchanged by β-CD, indicating that β-CD is unable to remove sphingomyelin or complexed Chol. However, β-CD removed all uncomplexed Chol from bilayers composed of global molar ratios below the stable complexation ratio. The removal of Chol by β-CD was independent of the initial structure of the membranes as deposited, suggesting that asymmetric membranes homogenize by the exchange of molecules between leaflets. The interaction of β-CD with the aforementioned membranes was independent of the deposition surface pressure except for a symmetric 50:50 membrane deposited at 40 mN/m. The scattering from 50:50 bilayers with higher packing densities (deposited at 40 mN/m) was unaffected by β-CD, suggesting that the removal of Chol can depend on both the composition and packing density of the membrane.     

Molecular dynamics simulations suggest that RNA three-way junctions can act as flexible RNA structural elements in the ribosome

We present extensive explicit solvent molecular dynamics analysis of three RNA three-way junctions (3WJs) from the large ribosomal subunit: the 3WJ formed by Helices 90–92 (H90–H92) of 23S rRNA; the 3WJ formed by H42–H44 organizing the GTPase associated center (GAC) of 23S rRNA; and the 3WJ of 5S rRNA. H92 near the peptidyl transferase center binds the 3′-CCA end of amino-acylated tRNA. The GAC binds protein factors and stimulates GTP hydrolysis driving protein synthesis. The 5S rRNA binds the central protuberance and A-site finger (ASF) involved in bridges with the 30S subunit. The simulations reveal that all three 3WJs possess significant anisotropic hinge-like flexibility between their stacked stems and dynamics within the compact regions of their adjacent stems. The A-site 3WJ dynamics may facilitate accommodation of tRNA, while the 5S 3WJ flexibility appears to be essential for coordinated movements of ASF and 5S rRNA. The GAC 3WJ may support large-scale dynamics of the L7/L12-stalk region. The simulations reveal that H42–H44 rRNA segments are not fully relaxed and in the X-ray structures they are bent towards the large subunit. The bending may be related to L10 binding and is distributed between the 3WJ and the H42–H97 contact.     

Harnessing the lysosome-dependent antitumor activity of phenothiazines in human small cell lung cancer

Phenothiazines are a family of heterocyclic compounds whose clinical utility includes treatment of psychiatric disorders as well as chemotherapy-induced emesis. Various studies have demonstrated that these compounds possess cytotoxic activities in tumor cell lines of different origin. However, there is considerable confusion regarding the molecular basis of phenothiazine-induced cell death. Lung cancer (LC) remains one of the most prevalent and deadly malignancies worldwide despite considerable efforts in the development of treatment strategies, especially new targeted therapies. In this work, we evaluated the potential utility of phenothiazines in human LC. We show that phenothiazines as single treatment decreased cell viability and induced cell death preferentially in small cell lung carcinoma (SCLC) over non small cell lung carcinoma (NSCLC) cell lines. Sensitivity to phenothiazines was not correlated with induction of apoptosis but due to phenothiazine-induced lysosomal dysfunction. Interestingly, the higher susceptibility of SCLC cells to phenothiazine-induced cell death correlated with an intrinsically lower buffer capacity in response to disruption of lysosomal homeostasis. Importantly, this effect in SCLC occurred despite mutation in p53 and was not influenced by intrinsic sensitivity/resistance toward conventional chemotherapeutic agents. Our data thus uncovered a novel context-dependent activity of phenothiazines in SCLC and suggest that phenothiazines could be considered as a treatment regimen of this disease, however, extended cell line analyses as well as in vivo studies are needed to make such conclusion.