The potential of mass rearing of Monoksa dorsiplana (Pteromalidae) a native gregarious ectoparasitoid of Pseudopachymeria spinipes (Bruchidae) in South America

In Chile and Uruguay, the gregarious Pteromalidae (Monoksa dorsiplana) has been discovered emerging from seeds of the persistent pods of Acacia caven attacked by the univoltin bruchid Pseudopachymeria spinipes. We investigated the potential for mass rearing of this gregarious ectoparasitoid on an alternative bruchid host, Callosobruchus maculatus, to use it against the bruchidae of native and cultured species of Leguminosea seeds in South America.The mass rearing of M. dorsiplana was carried out in a population cage where the density of egg-laying females per infested seed was increased from 1:1 on the first day to 5:1 on the last (fifth) day. Under these experimental conditions egg-clutch size per host increased, and at the same time the mortality of eggs laid also increased. The density of egg-laying females influenced the sex ratio which tended towards a balance of sons and daughters, in contrast to the sex ratio of a single egg-laying female per host (1 son to 7 daughters). The mean weight of adults emerging from a parasitized host was negatively correlated with the egg-clutch size, i.e. as egg-clutch size increased, adult weight decreased.All these results show that mass rearing of the gregarious ectoparasitoid M. dorsiplana was possible under laboratory conditions on an alternative bruchid host C. maculatus. As M. dorsiplana is a natural enemy of larval and pupal stages of bruchidae, the next step was to investigate whether the biological control of bruchid C. maculatus was possible in an experimental structure of stored beans.     

Improved agarose gel assay for quantification of growth factor-induced cell motility

Cell chemotaxis is frequently required in normal or pathological situations such as invasion, metastasis, and tumor angiogenesis and may involve many different cell types. At present, no device can simultaneously (i) make morphological observations, (ii) quantify cell migration, (iii) test multiple chemoattracting gradients, and (iv) analyze cell-cell interactions. We developed an agarose-based assay to address these questions. Two glass molds were designed, around which agarose gel could be poured to form specific well shapes. Using a vital nuclear stain (Hoechst 33258), we characterized the migration profile of adherent or suspension cells. Cells could be observed during the entire migration process. We were able to follow cells moving toward chemoattractants or being repulsed by other molecules, and we could estimate average migration speed. Using this inexpensive assay, we were able to obtain precise, reproducible results concerning the chemotactic behavior of different cell types. The resulting data differentiated between chemokinetic and chemotactic movement. Chemotactic potencies could be compared using different criteria, such as the number of attracted cells, induced speed, and morphological aspect. This improved agarose assay appears to be a reliable and inexpensive alternative to other available chemotaxis study tools.     

Mechanism of hcnA mRNA recognition in the Gac/Rsm signal transduction pathway of Pseudomonas fluorescens

In the plant-beneficial bacterium Pseudomonas fluorescens CHA0, the expression of antifungal exoproducts is controlled by the GacS/GacA two-component system. Two RNA binding proteins (RsmA, RsmE) ensure effective translational repression of exoproduct mRNAs. At high cell population densities, GacA induces three small RNAs (RsmX, RsmY, RsmZ) which sequester both RsmA and RsmE, thereby relieving translational repression. Here we systematically analyse the features that allow the RNA binding proteins to interact strongly with the 5' untranslated leader mRNA of the P. fluorescens hcnA gene (encoding hydrogen cyanide synthase subunit A). We obtained evidence for three major RsmA/RsmE recognition elements in the hcnA leader, based on directed mutagenesis, RsmE footprints and toeprints, and in vivo expression data. Two recognition elements were found in two stem-loop structures whose existence in the 5' leader region was confirmed by lead(II) cleavage analysis. The third recognition element, which overlapped the hcnA Shine-Dalgarno sequence, was postulated to adopt either an open conformation, which would favour ribosome binding, or a stem-loop structure, which may form upon interaction with RsmA/RsmE and would inhibit access of ribosomes. Effective control of hcnA expression by the Gac/Rsm system appears to result from the combination of the three appropriately spaced recognition elements.     

Circulating tumor cells: detection, molecular profiling and future prospects

Disseminated malignancy is responsible for the vast majority of cancer-related deaths. During this process, circulating tumor cells (CTC) are generated, spread from the primary tumor, colonize distant organs and lead to overt metastatic disease. CTC are essential for establishing metastasis; however, they are not sufficient as this process is highly inefficient and most will fail to grow in target sites. Several CTC die during migration while others remain dormant for several years and very few grow into macrometastases. CTC have been well documented in the bloodstream of cancer patients; however, the clinical relevance of this detection is still the subject of controversies and their biology is poorly understood. Indeed, available markers fail to distinguish between subgroups of CTC, and several current methods lack sensitivity, specificity or reproducibility in CTC characterization and detection. The advent of more precise technologies is renewing the interest in CTC biology. We will review herein recent findings on CTC biology, on the role of host-tumor interactions in CTC shedding and implantation, available methods of CTC detection and future perspectives for the molecular characterization of the CTC subset(s) responsible for the development of metastasis. Ultimately, understanding CTC biology and host-tumor 'complementarities' will help define metastasis-related biomarkers providing formidable and tailored novel therapeutic targets.     

Modification of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) film by chemical graft copolymerization

The graft copolymerization of 2-hydroxyethylmethacrylate (HEMA) onto poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBHV) films has been investigated. The graft copolymerization was conducted in aqueous media using benzoyl peroxide (BPO) as chemical initiator. PHBHV films were prepared by solvent casting. Different parameters affecting the graft yield were studied such as monomer concentration, initiator concentration, and reaction time. The extent of grafting has been modulated by the preparation conditions, in particular the concentration of HEMA. However, it is interesting to note that the initiator concentration had only a slight influence on the graft yield. Characterization of the grafted PHBHV films assumed that the graft copolymerization not only occurred on the film surface but also took place into the film bulk. Differential scanning calorimetry showed that crystallinity dramatically decreased with increasing graft yield, indicating that graft copolymerization hindered the crystallization process. Wettability has been obviously improved by grafting a hydrophilic monomer such as HEMA for high graft yield (>130%).     

The shifting influence of drought and heat stress for crops in northeast Australia

Characterization of drought environment types (ETs) has proven useful for breeding crops for drought‐prone regions. Here, we consider how changes in climate and atmospheric carbon dioxide (CO₂) concentrations will affect drought ET frequencies in sorghum and wheat systems of northeast Australia. We also modify APSIM (the Agricultural Production Systems Simulator) to incorporate extreme heat effects on grain number and weight, and then evaluate changes in the occurrence of heat‐induced yield losses of more than 10%, as well as the co‐occurrence of drought and heat. More than six million simulations spanning representative locations, soil types, management systems, and 33 climate projections led to three key findings. First, the projected frequency of drought decreased slightly for most climate projections for both sorghum and wheat, but for different reasons. In sorghum, warming exacerbated drought stresses by raising the atmospheric vapor pressure deficit and reducing transpiration efficiency (TE), but an increase in TE due to elevated CO₂ more than offset these effects. In wheat, warming reduced drought stress during spring by hastening development through winter and reducing exposure to terminal drought. Elevated CO₂ increased TE but also raised radiation‐use efficiency and overall growth rates and water use, thereby offsetting much of the drought reduction from warming. Second, adding explicit effects of heat on grain number and grain size often switched projected yield impacts from positive to negative. Finally, although average yield losses associated with drought will remain generally higher than that for heat stress for the next half century, the relative importance of heat is steadily growing. This trend, as well as the likely high degree of genetic variability in heat tolerance, suggests that more emphasis on heat tolerance is warranted in breeding programs. At the same time, work on drought tolerance should continue with an emphasis on drought that co‐occurs with extreme heat.