Development and field evaluation of liquid inoculants with native Bradyrhizobial strains for peanut production

A critical process in the leguminous crops cycles is biological nitrogen fixation (BNF). Application of inoculants with N fixing bacteria is economically and environmentally favourable. The aim of this work was to select competitive native peanut microsymbionts, evaluate their survival in inoculant support and assess their impact on peanut ( Arachis hypogaea L.) production under field conditions at C\uf3rdoba province in Argentina. The efficient N fixing Bradyrhizobium sp. J-81 and Bradyrhizobium sp. J-237, previously obtained from peanut nodules in the region of Cordoba, Argentina, were evaluated. In microcosm assays, plants inoculated with these isolates demonstrated better symbiotic parameters than those inoculated with reference strains. Different bacterial growth media and inoculant stabiliser solutions were evaluated. Balanced medium and arabic gum stabilising solution had optimal bacterial growth and the highest bacterial concentration and viability, respectively. Inoculation with either inoculants resulted in 44% greater peanut pod yield at Pizarro compared to the non-inoculated plants, although no significant differences were found with respect to commercial inoculants treatments.La fixation biologique de l\u2019Azote (FBA) est un processus important dans le cycle de vie des l\ue9gumineuses. L\u2019application d\u2019inoculum de bact\ue9ries fixatrices d\u2019azote est favorable au double plan \ue9conomique et environnemental. Le but de cette \ue9tude \ue9tait de s\ue9lectionner des bact\ue9ries symbiotiques de l\u2019arachide natives et comp\ue9titives, \ue9valuer leur temps de survie dans support d\u2019inoculum et \ue9valuer leur impact sur la production en plein champ de l\u2019arachide ( Arachis hypogaea L.) dans la province de C\uf3rdoba en Argentine. Les bact\ue9ries fixatrices d\u2019azote Bradyrhizobium sp. J-81 et Bradyrhizobium sp. J-237, extraites de nodules collect\ue9s sur des plants cultiv\ue9s dans la r\ue9gion de Cordoba en Argentina, ont \ue9t\ue9 \ue9valu\ue9es. Dans des essais de microcosme, des plants inocul\ue9s avec ces isolats ont exhib\ue9s de meilleurs param\ue8tres symbiotiques que les plants non inocul\ue9s. Diff\ue9rents m\ue9dia de culture bact\ue9rienne et supports inoculums ont \ue9t\ue9 test\ue9s. Medium mixte et solution stabilis\ue9e \ue0 la gomme arabique ont respectivement exhib\ue9s la croissance optimale des bact\ue9ries et la meilleure conservation et viabilit\ue9 des bact\ue9ries. L\u2019application de n\u2019importe quel inoculum produisit 44% plus de rendement en gousses d\u2019arachides \ue0 Pizarro par rapport aux plantes non-inocul\ue9es, et ceci bien qu\u2019aucune diff\ue9rence significative n\u2019a \ue9t\ue9 observ\ue9e en comparaison avec les traitements \ue0 l\u2019inoculum du commerce

Cell Cycle-Dependent Microtubule-Based Dynamic Transport of Cytoplasmic Dynein in Mammalian Cells

BACKGROUND:Cytoplasmic dynein complex is a large multi-subunit microtubule (MT)-associated molecular motor involved in various cellular functions including organelle positioning, vesicle transport and cell division. However, regulatory mechanism of the cell-cycle dependent distribution of dynein has not fully been understood. METHODOLOGY/PRINCIPAL FINDINGS:Here we report live-cell imaging of cytoplasmic dynein in HeLa cells, by expressing multifunctional green fluorescent protein (mfGFP)-tagged 74-kDa intermediate chain (IC74). IC74-mfGFP was successfully incorporated into functional dynein complex. In interphase, dynein moved bi-directionally along with MTs, which might carry cargos such as transport vesicles. A substantial fraction of dynein moved toward cell periphery together with EB1, a member of MT plus end-tracking proteins (+TIPs), suggesting +TIPs-mediated transport of dynein. In late-interphase and prophase, dynein was localized at the centrosomes and the radial MT array. In prometaphase and metaphase, dynein was localized at spindle MTs where it frequently moved from spindle poles toward chromosomes or cell cortex. +TIPs may be involved in the transport of spindle dyneins. Possible kinetochore and cortical dyneins were also observed. CONCLUSIONS AND SIGNIFICANCE:These findings suggest that cytoplasmic dynein is transported to the site of action in preparation for the following cellular events, primarily by the MT-based transport. The MT-based transport may have greater advantage than simple diffusion of soluble dynein in rapid and efficient transport of the limited concentration of the protein

Roles of Dynein and Dynactin in Early Endosome Dynamics Revealed Using Automated Tracking and Global Analysis

Microtubule-dependent movement is crucial for the spatial organization of endosomes in most eukaryotes, but as yet there has been no systematic analysis of how a particular microtubule motor contributes to early endosome dynamics. Here we tracked early endosomes labeled with GFP-Rab5 on the nanometer scale, and combined this with global, first passage probability (FPP) analysis to provide an unbiased description of how the minus-end microtubule motor, cytoplasmic dynein, supports endosome motility. Dynein contributes to short-range endosome movement, but in particular drives 85–98% of long, inward translocations. For these, it requires an intact dynactin complex to allow membrane-bound p150Glued to activate dynein, since p50 over-expression, which disrupts the dynactin complex, inhibits inward movement even though dynein and p150Glued remain membrane-bound. Long dynein-dependent movements occur via bursts at up to ∼8 µms−1 that are linked by changes in rate or pauses. These peak speeds during rapid inward endosome movement are still seen when cellular dynein levels are 50-fold reduced by RNAi knock-down of dynein heavy chain, while the number of movements is reduced 5-fold. Altogether, these findings identify how dynein helps define the dynamics of early endosomes

Quantitative Analysis of Lipid Droplet Fusion: Inefficient Steady State Fusion but Rapid Stimulation by Chemical Fusogens

Lipid droplets (LDs) are dynamic cytoplasmic organelles containing neutral lipids and bounded by a phospholipid monolayer. Previous studies have suggested that LDs can undergo constitutive homotypic fusion, a process linked to the inhibitory effects of fatty acids on glucose transporter trafficking. Using strict quantitative criteria for LD fusion together with refined light microscopic methods and real-time analysis, we now show that LDs in diverse cell types show low constitutive fusogenic activity under normal growth conditions. To investigate the possible modulation of LD fusion, we screened for agents that can trigger fusion. A number of pharmacological agents caused homotypic fusion of lipid droplets in a variety of cell types. This provided a novel cell system to study rapid regulated fusion between homotypic phospholipid monolayers. LD fusion involved an initial step in which the two adjacent membranes became continuous (<10 s), followed by the slower merging (100 s) of the neutral lipid cores to produce a single spherical LD. These fusion events were accompanied by changes to the LD surface organization. Measurements of LDs undergoing homotypic fusion showed that fused LDs maintained their initial volume, with a corresponding decrease in surface area suggesting rapid removal of membrane from the fused LD. This study provides estimates for the level of constitutive LD fusion in cells and questions the role of LD fusion in vivo. In addition, it highlights the extent of LD restructuring which occurs when homotypic LD fusion is triggered in a variety of cell types

Foregut caustic injuries: Results of the world society of emergency surgery consensus conference

Introduction: Lesions of the upper digestive tract due to ingestion of caustic agents still represent a major medical and surgical emergency worldwide. The work-up of these patients is poorly defined and no clear therapeutic guidelines are available. Purpose of the study: The aim of this study was to provide an evidence-based international consensus on primary and secondary prevention, diagnosis, staging, and treatment of this life-threatening and potentially disabling condition. Methods: An extensive literature search was performed by an international panel of experts under the auspices of the World Society of Emergency Surgery (WSES). The level of evidence of the screened publications was graded using the Oxford 2011 criteria. The level of evidence of the literature and the main topics regarding foregut caustic injuries were discussed during a dedicated meeting in Milan, Italy (April 2015), and during the 3rd Annual Congress of the World Society of Emergency Surgery in Jerusalem, Israel (July 2015). Results: One-hundred-forty-seven full papers which addressed the relevant clinical questions of the research were admitted to the consensus conference. There was an unanimous consensus on the fact that the current literature on foregut caustic injuries lacks homogeneous classification systems and prospective methodology. Moreover, the non-standardized definition of technical and clinical success precludes any accurate comparison of therapeutic modalities. Key recommendations and algorithms based on expert opinions, retrospective studies and literature reviews were proposed and approved during the final consensus conference. The clinical practice guidelines resulting from the consensus conference were approved by the WSES council. Conclusions: The recommendations emerging from this consensus conference, although based on a low level of evidence, have important clinical implications. A world registry of foregut caustic injuries could be useful to collect a homogeneous data-base for prospective clinical studies that may help improving the current clinical practice guidelines

Protein quality control: the who’s who, the where’s and therapeutic escapes

In cells the quality of newly synthesized proteins is monitored in regard to proper folding and correct assembly in the early secretory pathway, the cytosol and the nucleoplasm. Proteins recognized as non-native in the ER will be removed and degraded by a process termed ERAD. ERAD of aberrant proteins is accompanied by various changes of cellular organelles and results in protein folding diseases. This review focuses on how the immunocytochemical labeling and electron microscopic analyses have helped to disclose the in situ subcellular distribution pattern of some of the key machinery proteins of the cellular protein quality control, the organelle changes due to the presence of misfolded proteins, and the efficiency of synthetic chaperones to rescue disease-causing trafficking defects of aberrant proteins

The role of open abdomen in non-trauma patient : WSES Consensus Paper

The open abdomen (OA) is defined as intentional decision to leave the fascial edges of the abdomen un-approximated after laparotomy (laparostomy). The abdominal contents are potentially exposed and therefore must be protected with a temporary coverage, which is referred to as temporal abdominal closure (TAC). OA use remains widely debated with many specific details deserving detailed assessment and clarification. To date, in patients with intra-abdominal emergencies, the OA has not been formally endorsed for routine utilization; although, utilization is seemingly increasing. Therefore, the World Society of Emergency Surgery (WSES), Abdominal Compartment Society (WSACS) and the Donegal Research Academy united a worldwide group of experts in an international consensus conference to review and thereafter propose the basis for evidence-directed utilization of OA management in non-trauma emergency surgery and critically ill patients. In addition to utilization recommendations, questions with insufficient evidence urgently requiring future study were identified.Peer reviewe

Russell bodies as a model of ER storage diseases

Protein accumulation represents the consequence of an altered cellular homeostasis leading to an imbalance between synthesis and disposal. Cells adopt a number of strategies to cope with this imbalance: through the Unfolded Protein Response (UPR), intracellular chaperons are increased to facilitate protein folding, and the Endoplasmic Reticulum (ER) expands to accommodate increased concentration of ER resident proteins. The induction of autophagy is also a strategy adopted to face the synthesis of aberrant proteins. Our cellular model of protein accumulation regards mutant immunoglobulin: Ig-\ub5 chain that lacks the first constant domain (\ub5 06CH1 chain). These aberrant chains can neither exit from, nor are efficiently degraded in the ER. As a consequence they accumulate generating dilated cisternae known as Russell bodies. Russell bodies are frequently detected in lymphoproliferative diseases, especially in disorders of secretory B cells. Condensation of aberrant \ub5 06CH1 chains can occur in different sub-cellular locations: when Ig-L chains are produced detergent insoluble aggregates form in the rough ER; without L chains, aggregation occurs in ERGIC compartment. We are interested to define whether and which cellular mechanisms are active or are impaired by the synthesis of aberrant Ig-\ub5 chains assaying: i.e. ER stress, ER expansion, Autophagy modulation in our inducible cellular model (Hela-tet off) of Russell bodies formation. Our aim is to define if Russell bodies structures are a cell defence mechanism against proteotoxicit