Survey and phylogenetic analysis of culturable microbes in the oral secretions of three bark beetle species

In a recent study, we reported a previously undescribed behavior in which a bark beetle exuded oral secretions containing bacteria that have antifungal properties, and hence defend their galleries against pervasive antagonistic Hyphomycete fungi. Actinobacteria, a group known for their antibiotic properties, were the most effective against fungi that invade the spruce beetle galleries. In the present study, we describe the isolation and identification of microorganisms from oral secretions of three bark beetles (Coleoptera: Curculionidae: Scolytinae): the spruce beetle, Dendroctonus rufipennis Kirby, the mountain pine beetle, Dendroctonus ponderosae Hopkins, and the pine engraver, Ips pini Say. Bacteria isolated from these three species span the major bacterial classes α-, β-, and γ-Proteobacteria, Firmicutes, Bacteroidetes, and Actinobacteria, except for D. ponderosae , which yielded no α-proteobacteria or Bacteroidetes isolates. Spruce beetles and pine engraver beetles had similar numbers of α-proteobacteria isolates, but pine engravers yielded twice as many Bacteroidetes isolates as spruce beetles. In contrast, mountain pine beetles yielded more isolates in the β- and γ-proteobacteria than spruce beetles and pine engravers. The highest percentage of Actinobacteria was obtained from spruce beetles, followed by pine engravers and mountain pine beetles. All of the fungal isolates obtained from the three beetle species were Ascomycetes. The greatest fungal diversity was obtained in spruce beetles, which had nine species, followed by pine engravers with five, and mountain pine beetles with one.     

Hetero-oligomerization of reggie-1/flotillin-2 and reggie-2/flotillin-1 is required for their endocytosis

Reggie-1/flotillin-2 and reggie-2/flotillin-1 are membrane raft associated proteins which have been implicated in growth factor signaling, phagocytosis, regulation of actin cytoskeleton and membrane trafficking. Membrane and raft association of reggies is mediated by myristoylation, palmitoylation and oligomerization. We have shown that upon EGF stimulation of cells, reggie-1 is tyrosine phosphorylated by Src kinase and endocytosed into late endosomes. Here we have analyzed the mechanism of the EGF-stimulated endocytosis of reggies in more detail and show that the Src-mediated phosphorylation of reggie-1 is not the driving force for endocytosis. However, hetero-oligomerization with reggie-2 is necessary for the translocation of reggie-1, which does not take place in the absence of reggie-2. In addition, the Y163F mutant of reggie-1, which is not capable of undergoing endocytosis, oligomerizes poorly with reggie-2. EGF stimulation results in changes in the size but not in the stoichiometry of the reggie hetero-oligomers, and reggie-1 oligomer size is decreased by knockdown of reggie-2. Based on our findings, we propose a model according to which reggie hetero-oligomers are dynamic, and changes in the size of the hetero-oligomers result in endocytosis of the complex from the plasma membrane.     

Genotypes of Brassica rapa respond differently to plant-induced variation in air CO2 concentration in growth chambers with standard and enhanced venting

Growth chambers allow measurement of phenotypic differences among genotypes under controlled environment conditions. However, unintended variation in growth chamber air CO₂ concentration ([CO₂]) may affect the expression of diverse phenotypic traits, and genotypes may differ in their response to variation in [CO₂]. We monitored [CO₂] and quantified phenotypic responses of 22 Brassica rapa genotypes in growth chambers with either standard or enhanced venting. [CO₂] in chambers with standard venting dropped to 280 μmol mol^?1 during the period of maximum canopy development, ~80 μmol mol^?1 lower than in chambers with enhanced venting. The stable carbon isotope ratio of CO₂ in chamber air (δ¹³C_air) was negatively correlated with [CO₂], suggesting that photosynthesis caused observed [CO₂] decreases. Significant genotype × chamber-venting interactions were detected for 12 of 20 traits, likely due to differences in the extent to which [CO₂] changed in relation to genotypes’ phenology or differential sensitivity of genotypes to low [CO₂]. One trait, ¹³C discrimination (δ¹³C), was particularly influenced by unaccounted-for fluctuations in δ¹³C_air and [CO₂]. Observed responses to [CO₂] suggest that genetic variance components estimated in poorly vented growth chambers may be influenced by the expression of genes involved in CO₂ stress responses; genotypic values estimated in these chambers may likewise be misleading such that some mapped quantitative trait loci may regulate responses to CO₂ stress rather than a response to the environmental factor of interest. These results underscore the importance of monitoring, and where possible, controlling [CO₂].     

The role of p38α mitogen-activated protein kinase gene in the HELLP syndrome

Mitogen-activated protein kinase (MAPK) p38α was shown to be implicated in the organogenesis of the placenta, and such placental alteration is crucial for the development of hemolysis, elevated liver enzymes, and low platelets (HELLP) syndrome. We aimed to analyze for the first time human placental expression of MAPK p38α in pregnancies complicated by HELLP. The placental expression of MAPK p38α was investigated by semiquantitative polymerase chain reaction using cDNA extracted from placental tissue of 15 pregnancies with HELLP syndrome and 15 gestational age-matched controls. Seven patients with HELLP also had intrauterine fetal growth restriction (IUGR). In placenta from pregnancy complicated by HELLP, the expression of MAPK p38α is significantly decreased compared to the group with normal pregnancy (p < 0.001), while no difference was found between the HELLP and HELLP with IUGR subpopulations. Our study shows for the first time that MAPK p38α is expressed in the human placenta. Pregnancies with placental dysfunction and hypertensive complications are characterized by a significantly decreased expression of MAPK p38α. Our observations suggest that p38 MAPK signaling may be essential in placental angiogenesis and functioning.     

Evaluation of some biological parameters of Opuntia ficus indica. 2. Influence of seed supplemented diet on rats

The present research was undertaken to evaluate some biological parameters in rats fed with a supplemented diet with Opuntia ficus indica powder seeds. Feed intake and body weight of rats were measured every two days during nine weeks of treatment. Digestibility, feed conversion efficiency and protein efficiency ratio were determined. No difference in digestibility was noticed between the different diets. The results indicated a significant decrease in body weight of rats receiving a diet partially substituted with O. ficus indica powder seeds, probably due to a significant decrease in serum-free thyroxin (FT(4)) compared to the control group. In the treated group, a decrease of glucose concentration in blood and an increase of glycogen in liver and skeletal muscle were noticed. A significant increase in HDL-cholesterol was noted in the group receiving the supplemented diet with O. ficus indica powder seeds. These results suggest that O. ficus indica seeds can be used as a healthy food.     

Vascular endothelial growth factor-D activates VEGFR-3 expressed in osteoblasts inducing their differentiation

Vascular endothelial growth factor (VEGF)-D is a member of the VEGF family of angiogenic growth factors that recognizes and activates the vascular endothelial growth factor receptor (VEGFR)-2 and VEGFR-3 on blood and/or lymphatic vessels. We show that in the long bones of newborn mice, VEGF-D and VEGFR-3 are expressed in the osteoblasts of the growing plate. The treatment of primary human osteoblasts with recombinant VEGF-D induces the expression of osteocalcin and the formation of mineralized nodules in a dose-dependent manner. A monoclonal neutralizing antibody, anti-VEGF-D, or silencing of VEGFR-3 by lentiviral-mediated expression of VEGFR-3 small hairpin RNA affects VEGF-D-dependent osteocalcin expression and nodule formation. Moreover, in primary human osteoblasts, VEGF-D expression is under the control of VEGF, and inhibition of VEGF-D/VEGFR-3 signaling, by monoclonal antibodies or VEGFR-3 silencing, affects VEGF-dependent osteoblast differentiation. These experiments establish that VEGF-D/VEGFR-3 signaling plays a critical role in osteoblast maturation and suggest that VEGF-D is a downstream effector of VEGF in osteogenesis.     

How can a life cycle inventory parametric model streamline life cycle assessment in the wooden pallet sector?

Purpose

This study discusses the use of parameterization within the life cycle inventory (LCI) in the wooden pallet sector, in order to test the effectiveness of LCI parametric models to calculate the environmental impacts of similar products. Starting from a single case study, the objectives of this paper are (1) to develop a LCI parametric model adaptable to a range of wooden pallets, (2) to test this model with a reference product (non-reversible pallet with four-way blocks) and (3) to determine numerical correlations between the environmental impacts and the most significant LCI parameters; these correlations can be used to improve the design of new wooden pallets.

Methods

The conceptual scheme for defining the model is based on ISO14040-44 standards. First of all, the product system was defined identifying the life cycle of a generic wood pallet, as well as its life cycle stages. A list of independent and dependent parameters was used to describe the LCI flows of a generic wooden pallet. The LCI parametric model was applied to calculate the environmental impacts of the reference product, with regard to a selection of impact categories at midpoint level (climate change, human toxicity, particulate matter formation, agricultural land occupation, fossil depletion). The model was then applied to further 11 wooden pallets belonging to the same category.

Results and discussion

The definition of a LCI parametric model based on 31 independent parameters and 21 dependent parameters streamlined the data collection process, as the information required for fulfilling the LCI are standard information about the features of the wooden pallet and its manufacturing process. The contribution analysis on the reference product revealed that the most contributing life cycle stages are wood and nails extraction and manufacturing (positive value of environmental impact) and end-of-life (avoided impact). This result is driven by two parameters: mass of wood and average distance for transport of wood. Based on the results of the application of the LCI parametric model to the identified products, one parameter-based regression and one multiple non-linear regression allowed to define a correlation between the life cycle impact assessment (LCIA) category indicators considered and the most influencing parameters.

Conclusions

The definition of LCI parametric model in the wooden pallet sector can effectively be used for calculating the environmental impacts of products with different designs, as well as for obtaining a preliminary estimation of the life cycle environmental impacts of new products.     

Snow Surface Microbiome on the High Antarctic Plateau (DOME C)

The cryosphere is an integral part of the global climate system and one of the major habitable ecosystems of Earth''s biosphere. These permanently frozen environments harbor diverse, viable and metabolically active microbial populations that represent almost all the major phylogenetic groups. In this study, we investigated the microbial diversity in the surface snow surrounding the Concordia Research Station on the High Antarctic Plateau through a polyphasic approach, including direct prokaryotic quantification by flow cytometry and catalyzed reporter deposition fluorescence in situ hybridization (CARD-FISH), and phylogenetic identification by 16S RNA gene clone library sequencing and 454 16S amplicon pyrosequencing. Although the microbial abundance was low (<10³ cells/ml of snowmelt), concordant results were obtained with the different techniques. The microbial community was mainly composed of members of the Alpha-proteobacteria class (e.g. Kiloniellaceae and Rhodobacteraceae), which is one of the most well-represented bacterial groups in marine habitats, Bacteroidetes (e.g. Cryomorphaceae and Flavobacteriaceae) and Cyanobacteria. Based on our results, polar microorganisms could not only be considered as deposited airborne particles, but as an active component of the snowpack ecology of the High Antarctic Plateau.     

Separate and Combined Effects of DNMT and HDAC Inhibitors in Treating Human Multi-Drug Resistant Osteosarcoma HosDXR150 Cell Line

Understanding the molecular mechanisms underlying multi-drug resistance (MDR) is one of the major challenges in current cancer research. A phenomenon which is common to both intrinsic and acquired resistance, is the aberrant alteration of gene expression in drug-resistant cancers. Although such dysregulation depends on many possible causes, an epigenetic characterization is considered a main driver. Recent studies have suggested a direct role for epigenetic inactivation of genes in determining tumor chemo-sensitivity. We investigated the effects of the inhibition of DNA methyltransferase (DNMT) and hystone deacethylase (HDAC), considered to reverse the epigenetic aberrations and lead to the re-expression of de novo methylated genes in MDR osteosarcoma (OS) cells. Based on our analysis of the HosDXR150 cell line, we found that in order to reduce cell proliferation, co-treatment of MDR OS cells with DNMT (5-Aza-dC, DAC) and HDAC (Trichostatin A, TSA) inhibitors is more effective than relying on each treatment alone. In re-expressing epigenetically silenced genes induced by treatments, a very specific regulation takes place which suggests that methylation and de-acetylation have occurred either separately or simultaneously to determine MDR OS phenotype. In particular, functional relationships have been reported after measuring differential gene expression, indicating that MDR OS cells acquired growth and survival advantage by simultaneous epigenetic inactivation of both multiple p53-independent apoptotic signals and osteoblast differentiation pathways. Furthermore, co-treatment results more efficient in inducing the re-expression of some main pathways according to the computed enrichment, thus emphasizing its potential towards representing an effective therapeutic option for MDR OS.     

Construction of new stable strain over-expressing the glucose isomerase of the Streptomyces sp. SK strain

In order to over express the xylA gene of Streptomyces sp. SK strain, it was cloned under the control of the constitutive ermE-up promoter. This construct was integrated through site-specific recombination process into the chromosome of a Streptomyces violaceoniger glucose isomerase deficient strain using the non-replicative vector pTS55. The resulting CBS4 strain shows a perfect stability in the absence of selection pressure. Its glucose isomerase activity was about four and nine-fold greater, than that obtained from Streptomyces sp. SK, respectively fully induced or not by xylose.