Generation of thermo-sensitive allele of the TPR like protein Nup211by PCR mutagenesis

Motivation: TPR proteins are conserved large coiled-coil proteins that localize at the nucleoplasmic side of the nuclear pore complex and participate in multiple aspects of DNA metabolism. The protein Nup211, fission yeast homolog of Mlp1/Mlp2/Tpr, participate in the mRNA export and is essential for vegetative growth. The aim of this work is to create a collection of thermo-sensitive alleles of nup211.Methods: To create the collection, we have generated a new strain with the nup211 gen tagged with GFP at the amino terminal extreme and confirmed by fluorescent microscopy that the protein Nup211 localized in the nuclear envelop. Then, we have carried out a Taq PCR-based Random Mutagenesis with reduced concentration of dATP. The PCR products were transformed into a wild type strain to generate conditional mutants. The transformants obtained whose growth was impaired at 36ºC were preselected as thermo-sensitive mutants. To confirm the growth deficiency of these clones, a drop assay was performed and the best candidates were selected. These thermo-sensitive mutants were cultivated at 25ºC as well as 36ºC and both cultures were subjected to various experiments in order to study any changes in the localization of Nup211.Results: Up to now, we have demonstrated by fluorescent microscopy that the thermo-sensitive mutants show a modified nuclear distribution of Nup211 and different cellular phenotype, suggesting that the differents clones might represent differents nup211 thermo-sensitive alleles. These alleles are going to be subjected to various experiments to clarify the role of the protein in the mRNA export

The MicroBioDiverSar Project: Exploring the Microbial Biodiversity in Ex Situ Collections of Sardinia

In the last decades, biodiversity preservation has gained growing attention and many strategies, laws and regulations have been enacted by governments with this purpose. The Micro-BioDiverSar (MBDS) project, the first one regarding microbiological resources, funded by the Italian Minister of Agricultural, Food and Forestry Policies (Mipaaf) through the Law 194/2015, was aimed at surveying, cataloguing, and managing the microbial resources and the related information of three Sardinian collections (Agris BNSS, Uniss, and Unica). While microorganisms were reordered and inventoried, a federated database, accessible via the web, was designed by the bioinformatician of Ospedale Policlinico San Martino of Genova, according to both international standards and laboratory needs. The resulting MBDS collection boasts a great richness of microbial resources. Indeed, over 21,000 isolates, belonging to over 200 species of bacteria, yeasts, and filamentous fungi isolated from different matrices, mainly food, of animal and vegetable origin, collected in over 50 years, were included in the database. Currently, about 2000 isolates, belonging to 150 species, are available online for both the scientific community and agri-food producers. The huge work done allowed one to know the consistency and the composition of most of the patrimony of the Sardinian microbial collections. Furthermore, the MBDS database has been proposed as a model for other Italian collections that, as the MBDS partners, are part of the Joint Research UnitMIRRI-IT Italian collections network, with the aim of overcoming fragmentation, facing sustainability challenges, and improving the quality of the management of the collections

Do best-selected strains perform table olive fermentation better than undefined biodiverse starters? A comparative study

Twenty-seven Lactobacillus pentosus strains, and the undefined starter for table olives from which they were isolated, were characterised for their technological properties: tolerance to low temperature, high salt concentration, alkaline pH, and olive leaf extract; acidifying ability; oleuropein degradation; hydrogen peroxide and lactic acid production. Two strains with appropriate technological properties were selected. Then, table olive fermentation in vats, with the original starter, the selected strains, and without starter (spontaneous fermentation) were compared. Starters affected some texture profile parameters. The undefined culture resulted in the most effective Enterobacteriaceae reduction, acidification and olive debittering, while the selected strains batch showed the lowest antioxidant activity. Our results show that the best candidate strains cannot guarantee better fermentation performance than the undefined biodiverse mix from which they originate

Fear of falling and postural reactivity in patients with glaucoma

Purpose To investigate the relationship between postural metrics obtained by dynamic visual stimulation in a virtual reality environment and the presence of fear of falling in glaucoma patients. Methods This cross-sectional study included 35 glaucoma patients and 26 controls that underwent evaluation of postural balance by a force platform during presentation of static and dynamic visual stimuli with head-mounted goggles (Oculus Rift). In dynamic condition, a peripheral translational stimulus was used to induce vection and assess postural reactivity. Standard deviations of torque moments (SDTM) were calculated as indicative of postural stability. Fear of falling was assessed by a standardized questionnaire. The relationship between a summary score of fear of falling and postural metrics was investigated using linear regression models, adjusting for potentially confounding factors. Results Subjects with glaucoma reported greater fear of falling compared to controls (-0.21 vs. 0.27P = 0.039). In glaucoma patients, postural metrics during dynamic visual stimulus were more associated with fear of falling (R-2 = 18.8%P = 0.001) than static (R-2 = 3.0%P = 0.005) and dark field (R-2 = 5.7%P = 0.007) conditions. In the univariable model, fear of falling was not significantly associated with binocular standard perimetry mean sensitivity (P = 0.855). In the multivariable model, each 1 Nm larger SDTM in anteroposterior direction during dynamic stimulus was associated with a worsening of 0.42 units in the fear of falling questionnaire score (P = 0.001). Conclusion In glaucoma patients, postural reactivity to a dynamic visual stimulus using a virtual reality environment was more strongly associated with fear of falling than visual field testing and traditional balance assessment.National Institutes of Health/National Eye Institute [EY021818]Brazilian National Council for Scientific and Technological Development (CNPq) [233829/2014-8]Alcon Laboratories (Fort Worth, TX)Bausch & Lomb (Garden City, NY)Carl Zeiss Meditec (Jena, Germany)Heidelberg Engineering (Heidelberg, Germany)Merck (White House Station, NJ)Allergan (Irvine, CA)Sensimed (Lausanne, Switzerland)Topcon (Livermore, CA)Reichert (Dewey, NY)National Eye Institute (Bethesda, MD)Novartis (Basel, Switzerland)nGoggle (San Diego, CA)Duke Univ, Duke Eye Ctr, Durham, NC 27708 USADuke Univ, Dept Ophthalmol, Durham, NC 27708 USAUniv Fed Sao Paulo, Dept Ophthalmol & Vis Sci, Sao Paulo, BrazilUniv Calif San Diego, Dept Ophthalmol, La Jolla, CA 92093 USAUniv Fed Sao Paulo, Dept Ophthalmol & Vis Sci, Sao Paulo, BrazilCNPq [233829/2014-8]Web of Scienc

A Combined EM and Visual Tracking Probabilistic Model for Robust Mosaicking: Application to Fetoscopy

Twin-to-Twin Transfusion Syndrome (TTTS) is a progressive pregnancy complication in which inter-twin vascular connections in the shared placenta result in a blood flow imbalance between the twins. The most effective therapy is to sever these connections by laser photo-coagulation. However, the limited field of view of the fetoscope hinders their identification. A potential solution is to augment the surgeon’s view by creating a mosaic image of the placenta. State-of-the-art mosaicking methods use feature-based ap- proaches, which have three main limitations: (i) they are not robust against corrupt data e.g. blurred frames, (ii) tem- poral information is not used, (iii) the resulting mosaic suf- fers from drift. We introduce a probabilistic temporal model that incorporates electromagnetic and visual tracking data to achieve a robust mosaic with reduced drift. By assuming planarity of the imaged object, the nRT decomposition can be used to parametrize the state vector. Finally, we tackle the non-linear nature of the problem in a numerically stable manner by using the Square Root Unscented Kalman Filter. We show an improvement in performance in terms of robustness as well as a reduction of the drift in comparison to state-of-the-art methods in synthetic, phantom and ex vivo datasets

Dynamic constriction and fission of endoplasmic reticulum membranes by reticulon

The endoplasmic reticulum (ER) is a continuous cell-wide membrane network. Network formation has been associated with proteins producing membrane curvature and fusion, such as reticulons and atlastin. Regulated network fragmentation, occurring in different physiological contexts, is less understood. Here we find that the ER has an embedded fragmentation mechanism based upon the ability of reticulon to produce fission of elongating network branches. In Drosophila, Rtnl1-facilitated fission is counterbalanced by atlastin-driven fusion, with the prevalence of Rtnl1 leading to ER fragmentation. Ectopic expression of Drosophila reticulon in COS-7 cells reveals individual fission events in dynamic ER tubules. Consistently, in vitro analyses show that reticulon produces velocity-dependent constriction of lipid nanotubes leading to stochastic fission via a hemifission mechanism. Fission occurs at elongation rates and pulling force ranges intrinsic to the ER, thus suggesting a principle whereby the dynamic balance between fusion and fission controlling organelle morphology depends on membrane motility

Soil-transmitted helminth infections and nutritional indices among Filipino schoolchildren

BACKGROUND: Soil-transmitted helminth (STH) infections are still prevalent among schoolchildren in the Philippines. We evaluated the risk factors associated with STH and the relationship between STH and nutritional indices among schoolchildren aged 9-10 years in Laguna province, the Philippines. METHODS: We used the baseline data from 40 schools enrolled in a randomised controlled trial of the Magic Glasses Philippines health education package. Data on demographic and socio-economic variables, and STH related knowledge, attitudes and practices, were obtained through a questionnaire. Stool samples were collected and assessed for STH egg presence using the Kato-Katz technique. Haemoglobin levels and height and weight of study participants were also determined. The generalized estimating equations approach was used to construct logistic regression models to assess STH-associated risk factors, and the association between any STH infection and anaemia, child stunting, wasting and being underweight. The trial is registered with the Australian New Zealand Clinical Trials Registry (ACTRN12616000508471). FINDINGS: Among 1,689 schoolchildren, the prevalence of any STH was 23%. The prevalence of anaemia, stunting, being underweight and wasting was 13%, 20.2%, 19% and 9.5%, respectively. Age, socio-economic status, rural/urban classification of schools and knowledge of STH were significant risk factors for acquiring a STH infection. Moreover, infections with any STH were significantly associated with stunting (P = <0.001) and being underweight (P = <0.003), but not wasting (P = 0.375) or anaemia (P = 0.462) after controlling for confounding covariates. CONCLUSION: The study findings emphasise the need for sustainable deworming in tandem with other measures such as the provision of health education, improvements in sanitation and hygiene, and nutritional programs in order to control STH infections and improve morbidity outcomes in schoolchildren. TRIAL REGISTRATION: Australian New Zealand Clinical Trials Registry (ACTRN12616000508471)

Condensation in models with factorized and pair-factorized stationary states

Non-equilibrium real-space condensation is a phenomenon in which a finite fraction of some conserved quantity (mass, particles, etc.) becomes spatially localised. We review two popular stochastic models of hopping particles that lead to condensation and whose stationary states assume a factorized form: the zero-range process and the misanthrope process, and their various modifications. We also introduce a new model - a misanthrope process with parallel dynamics - that exhibits condensation and has a pair-factorized stationary state.Comment: 15 pages, 2 figures submitted to J. Stat. Mec