Prevalence and impact of multidrug resistance in a cohort of patients admitted to emergency department for urinary tract-infections: The UTILY study, a prospective multicentre study.

Background And Aim: The aim of the present study was to evaluate the prevalence and to identify the independent predictors of multi-drug resistance among a cohort of patients admitted to emergency department for urinary tract infections (UTI), and to assess the impact of antimicrobial resistance on the clinical outcomes.

Methods: We conducted a prospective multicentre study enrolling all adult patients admitted to one of the eight emergency departments participating in the study with a microbiologically confirmed diagnosis of UTI from February 2023 to July 2024. The primary outcome evaluated was 30-day mortality; secondary outcomes included 7-day mortality and clinical response.

Facile Conversion and Optimization of Structured Illumination Image Reconstruction Code into the GPU Environment.

Superresolution, structured illumination microscopy (SIM) is an ideal modality for imaging live cells due to its relatively high speed and low photon-induced damage to the cells. The rate-limiting step in observing a superresolution image in SIM is often the reconstruction speed of the algorithm used to form a single image from as many as nine raw images. Reconstruction algorithms impose a significant computing burden due to an intricate workflow and a large number of often complex calculations to produce the final image.

Investigation on Flavescence Dorée in North-Western Italy Identifies Map-M54 (16SrV-D/Map-FD2) as the Only Phytoplasma Genotype in L. and Reveals the Presence of New Putative Reservoir Plants.

Flavescence dorée (FD) is the most important phytoplasma-associated disease of the grapevine yellows complex in Europe. Recent studies highlighted a great genetic diversity within FD phytoplasma (FDp) strains and demonstrated that their diffusion is not related exclusively to the pathosystem including L. and but involves additional vectors and reservoir plants.

Signaling Cross-Talk between Salicylic and Gentisic Acid in the ' Phytoplasma Solani' Interaction with Sangiovese Vines.

"Bois noir" disease associated with ' Phytoplasma solani' seriously compromises the production and survival of grapevines ( L.) in Europe. Understanding the plant response to phytoplasmas should help to improve disease control strategies.

Zero-order free holographic optical tweezers.

Holographic optical tweezers (HOTs) use spatial light modulators (SLM) to modulate light beams, thereby enabling the dynamic control of optical trap arrays with complex intensity and phase distributions. This has provided exciting new opportunities for cell sorting, microstructure machining, and studying single molecules. However, the pixelated structure of the SLM will inevitably bring up the unmodulated zero-order diffraction possessing an unacceptably large fraction of the incident light beam power.

Rapid, artifact-reduced, image reconstruction for super-resolution structured illumination microscopy.

Super-resolution structured illumination microscopy (SR-SIM) is finding increasing application in biomedical research due to its superior ability to visualize subcellular dynamics in living cells. However, during image reconstruction artifacts can be introduced and when coupled with time-consuming postprocessing procedures, limits this technique from becoming a routine imaging tool for biologists. To address these issues, an accelerated, artifact-reduced reconstruction algorithm termed joint space frequency reconstruction-based artifact reduction algorithm (JSFR-AR-SIM) was developed by integrating a high-speed reconstruction framework with a high-fidelity optimization approach designed to suppress the sidelobe artifact.

Disease Forecasting for the Rational Management of Grapevine Mildews in the Chianti Bio-District (Tuscany).

Downy and powdery mildews are major grapevine diseases. In organic viticulture, a few fungicides with protectant activities (copper and sulphur in particular) can be used, and their preventative application frequently leads to unneeded spraying. The adoption of an epidemiological disease forecasting model could optimise the timing of treatments and achieve a good level of disease protection.

Influence of Nitrogen on Grapevine Susceptibility to Downy Mildew.

Downy mildew, caused by the obligate parasite , is one of the most important threats to viticulture. The exploitation of resistant and susceptibility traits of grapevine is one of the most promising ways to increase the sustainability of disease management. Nitrogen (N) fertilization is known for influencing disease severity in the open field, but no information is available on its effect on plant-pathogen interaction.

Use of Dual Optical Tweezers and Microfluidics for Single-Molecule Studies.

Visual biochemistry is a powerful technique for observing the stochastic properties of single enzymes or enzyme complexes that are obscured in the averaging that takes place in bulk-phase studies. To achieve visualization, dual optical tweezers, where one trap is fixed and the other is mobile, are focused into one channel of a multi-stream microfluidic chamber positioned on the stage of an inverted fluorescence microscope. The optical tweezers trap single molecules of fluorescently labeled DNA and fluid flow through the chamber and past the trapped beads, stretches the DNA to B-form (under minimal force, i.

Characterization of Italian Plasmopara viticola populations for resistance to oxathiapiprolin.

Background: Oxathiapiprolin is a novel fungicide and the first of the piperidinyl-thiazole-isoxazoline class to be discovered. This fungicide has been reported to have high activity against Plasmopara viticola, the grapevine downy mildew agent, and other plant-pathogenic oomycetes. In this study, the baseline sensitivity of Italian P.

The Biochemical Mechanism of Fork Regression in Prokaryotes and Eukaryotes-A Single Molecule Comparison.

The rescue of stalled DNA replication forks is essential for cell viability. Impeded but still intact forks can be rescued by atypical DNA helicases in a reaction known as fork regression. This reaction has been studied at the single-molecule level using the DNA helicase RecG and, separately, using the eukaryotic SMARCAL1 enzyme.

Biocontrol Potential of Endophytic Plant-Growth-Promoting Bacteria against Phytopathogenic Viruses: Molecular Interaction with the Host Plant and Comparison with Chitosan.

Endophytic plant-growth-promoting bacteria (ePGPB) are interesting tools for pest management strategies. However, the molecular interactions underlying specific biocontrol effects, particularly against phytopathogenic viruses, remain unexplored. Herein, we investigated the antiviral effects and triggers of induced systemic resistance mediated by four ePGPB ( strain R8, strain R16, strain 255-7, and strain 260-02) against four viruses (Cymbidium Ring Spot Virus-CymRSV; Cucumber Mosaic Virus-CMV; Potato Virus X-PVX; and Potato Virus Y-PVY) on plants under controlled conditions and compared them with a chitosan-based resistance inducer product.

OB-fold Families of Genome Guardians: A Universal Theme Constructed From the Small β-barrel Building Block.

The maintenance of genome stability requires the coordinated actions of multiple proteins and protein complexes, that are collectively known as genome guardians. Within this broadly defined family is a subset of proteins that contain oligonucleotide/oligosaccharide-binding folds (OB-fold). While OB-folds are widely associated with binding to single-stranded DNA this view is no longer an accurate depiction of how these domains are utilized.

Real-Time On-Site Diagnosis of Quarantine Pathogens in Plant Tissues by Nanopore-Based Sequencing.

Rapid and sensitive assays for the identification of plant pathogens are necessary for the effective management of crop diseases. The main limitation of current diagnostic testing is the inability to combine broad and sensitive pathogen detection with the identification of key strains, pathovars, and subspecies. Such discrimination is necessary for quarantine pathogens, whose management is strictly dependent on genotype identification.

Insight into the biochemical mechanism of DNA helicases provided by bulk-phase and single-molecule assays.

There are multiple assays available that can provide insight into the biochemical mechanism of DNA helicases. For the first 22 years since their discovery, bulk-phase assays were used. These include gel-based, spectrophotometric, and spectrofluorometric assays that revealed many facets of these enzymes.

SSB Facilitates Fork-Substrate Discrimination by the PriA DNA Helicase.

Primosomal protein A (PriA) is a member of helicase SuperFamily 2. Its role is to reload the primosome onto resurrected replication forks resulting in the restart of the previously stalled DNA replication process. Single-stranded DNA-binding protein (SSB) plays a key role in mediating activities at replication forks and interacts both physically and functionally with PriA.

Multi-color structured illumination microscopy for live cell imaging based on the enhanced image recombination transform algorithm.

Structured illumination microscopy (SIM) has attracted considerable interest in super-resolution, live-cell imaging because of its low light dose and high imaging speed. Obtaining a high-quality reconstruction image in SIM depends on the precise determination of the parameters of the fringe illumination pattern. The image recombination transform (IRT) algorithm is superior to other algorithms in obtaining the precise initial phase without any approximation, which is promising to provide a considerable solution to address the difficulty of initial phase estimation at low-modulation-depth conditions.

RNAi of a Putative Grapevine Susceptibility Gene as a Possible Downy Mildew Control Strategy.

Downy mildew, caused by the oomycete , is one of the diseases causing the most severe economic losses to grapevine () production. To date, the application of fungicides is the most efficient method to control the pathogen and the implementation of novel and sustainable disease control methods is a major challenge. RNA interference (RNAi) represents a novel biotechnological tool with a great potential for controlling fungal pathogens.

The mechanism of action of the SSB interactome reveals it is the first OB-fold family of genome guardians in prokaryotes.

The single-stranded DNA binding protein (SSB) is essential to all aspects of DNA metabolism in bacteria. This protein performs two distinct, but closely intertwined and indispensable functions in the cell. SSB binds to single-stranded DNA (ssDNA) and at least 20 partner proteins resulting in their regulation.

High-yield purification of exceptional-quality, single-molecule DNA substrates.

Single-molecule studies involving DNA or RNA, require homogeneous preparations of nucleic acid substrates of exceptional quality. Over the past several years, a variety of methods have been published describing different purification methods but these are frustratingly inconsistent with variable yields even in the hands of experienced bench scientists. To address these issues, we present an optimized and straightforward, column-based approach that is reproducible and produces high yields of substrates or substrate components of exceptional quality.

Dynamics of the PriA Helicase at Stalled DNA Replication Forks.

The DNA helicase PriA is a key protein for restarting stalled DNA replication forks in bacteria. With 3' to 5' helicase activity, PriA is important in primosome assembly. We used atomic force microscopy (AFM) and specifically employed time-lapse AFM to visualize the interaction of PriA with two DNA substrates.

Rapid Image Reconstruction of Structured Illumination Microscopy Directly in the Spatial Domain.

Super-resolution structured illumination microscopy (SIM) routinely performs image reconstruction in the frequency domain using an approach termed frequency-domain reconstruction (FDR). Due to multiple Fourier transforms between the spatial and frequency domains, SIM suffers from low reconstruction speed, constraining its applications in real-time, dynamic imaging. To overcome this limitation, we developed a new method for SIM image reconstruction, termed spatial domain reconstruction (SDR).

In Vivo Binding of Single-Stranded DNA-Binding Protein to Stalled Replication Fork Helicases.

Understanding protein-protein interactions is key to unraveling protein function in vivo. Here we describe a dual/triple-plasmid system that enables co-expression of two, or three, recombinant proteins harboring different affinity tags in the same Escherichia coli cell. This novel protein expression system provides a platform to understand protein-protein interactions and enables researchers to study protein complex formation and in vivo localization.

Characterize the Interaction of the DNA Helicase PriA with the Stalled DNA Replication Fork Using Atomic Force Microscopy.

In bacteria, the restart of stalled DNA replication forks requires the DNA helicase PriA. PriA can recognize and remodel abandoned DNA replication forks, unwind DNA in the 3'-to-5' direction, and facilitate the loading of the helicase DnaB onto the DNA to restart replication. ssDNA-binding protein (SSB) is typically present at the abandoned forks, protecting the ssDNA from nucleases.