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Abstract X-ray crystallography is one of the leading tools to analyze the 3-D structure, and therefore, function of proteins and other biological macromolecules. Traditional methods of mounting individual crystals for X-ray diffraction analysis can be tedious and result in damage to fragile protein crystals. Furthermore, the advent of multi-crystal and serial crystallography methods explicitly require the mounting of larger numbers of crystals. To address this need, we have developed a device that facilitates the straightforward mounting of protein crystals for diffraction analysis, and that can be easily manufactured at scale. Inspired by grid-style devices that have been reported in the literature, we have developed an X-ray compatible microfluidic device that can be used to trap protein crystals in an array configuration, while also providing excellent optical transparency, a low X-ray background, and compatibility with the robotic sample handling and environmental controls used at synchrotron macromolecular crystallography beamlines. At the Stanford Synchrotron Radiation Lightsource (SSRL), these capabilities allow for fully remote-access data collection at controlled humidity conditions. Furthermore, we have demonstrated continuous manufacturing of these devices via roll-to-roll fabrication to enable cost-effective and efficient large-scale production.

Mayo Clinic Ambulance Service is testing a novel combination of technologies to enhance the ability to provide prehospital telemedicine connecting physicians with paramedics. Mayo Clinic Ambulance Service partnered with start-up company OPTAC-X to field test a novel head-mounted video camera connected with a satellite communications terminal to bring medical control emergency medicine physicians to the patient and paramedic by video. The authors believe this is the first report of a physician providing medical guidance to paramedics resuscitating an out-of-hospital cardiac arrest using these technologies.

Over the past two decades, serial X-ray crystallography has enabled the structure determination of a wide range of proteins. With the advent of X-ray free-electron lasers (XFELs), ever-smaller crystals have yielded high-resolution diffraction and structure determination. A crucial need to continue advancement is the efficient delivery of fragile and micrometre-sized crystals to the X-ray beam intersection. This paper presents an improved design of an all-polymer microfluidic `chip for room-temperature fixed-target serial crystallography that can be tailored to broadly meet the needs of users at either synchrotron or XFEL light sources. The chips are designed to be customized around different types of crystals and offer users a friendly, quick, convenient, ultra-low-cost and robust sample-delivery platform. Compared with the previous iteration of the chip [Gilbile et al. (2021), Lab Chip, 21, 4831-4845], the new design eliminates cleanroom fabrication. It has a larger imaging area to volume, while maintaining crystal hydration stability for both in situ crystallization or direct crystal slurry loading. Crystals of two model proteins, lysozyme and thaumatin, were used to validate the effectiveness of the design at both synchrotron (lysozyme and thaumatin) and XFEL (lysozyme only) facilities, yielding complete data sets with resolutions of 1.42, 1.48 and 1.70 Å, respectively. Overall, the improved chip design, ease of fabrication and high modifiability create a powerful, all-around sample-delivery tool that structural biologists can quickly adopt, especially in cases of limited sample volume and small, fragile crystals.

ABSTRACT: The success of a crop begins with the use of high-quality seeds, especially with regard to the physiological potential for germination and vigor, as well as genetic and sanitary purity, and is also fundamental for the transfer of technological innovations and genetic gains resulting from breeding works, whether traditional or the result of genetic engineering, to the field. New techniques have been studied in order to mitigate possible damage that may affect seeds until planting in the field. Thus, the objective of this study was to evaluate the effect of salicylic acid on the physiological and sanitary quality of high- and low-vigor maize seeds. High- and low-vigor seeds of the maize hybrid B2315PWU were subjected to soaking at salicylic acid concentrations of 0, 500, 1000, 1500, 2000, 2500, 3000, 3500 and 4000 uM (Micromolar). Evaluations of first count, germination, cold vigor, seedling length and mass, and sanity test were carried out. Salicylic acid doses up to 2000 uM improve germination potential in maize seeds. Treatment of maize seeds with salicylic acid may be a promising alternative for the control of Fusarium spp.

The physicochemical properties and oxidative stability of encapsulated fish oil in a matrix formed by the reaction of crude de-oiled soy lecithin and dextrose following a Maillard reaction path were investigated. The effects of sugar content in formula (4, 8 %) and temperature (25 °C, 80 °C, and 100 °C) on the moisture content, water activity, encapsulation efficiency (EE), and oxidative stability were studied. Oxidative stability was measured over a 90-day testing period at 25 °C. Samples achieved an encapsulation efficiency in the range 50–70 %, low moisture content (

PurposeDominant variants in the retinoic acid receptor beta (RARB) gene underlie a syndromic form of microphthalmia, known as MCOPS12, which is associated with other birth anomalies and global developmental delay with spasticity and/or dystonia. Here, we report 25 affected individuals with 17 novel pathogenic or likely pathogenic variants in RARB. This study aims to characterize the functional impact of these variants and describe the clinical spectrum of MCOPS12.MethodsWe used in vitro transcriptional assays and in silico structural analysis to assess the functional relevance of RARB variants in affecting the normal response to retinoids.ResultsWe found that all RARB variants tested in our assays exhibited either a gain-of-function or a loss-of-function activity. Loss-of-function variants disrupted RARB function through a dominant-negative effect, possibly by disrupting ligand binding and/or coactivators' recruitment. By reviewing clinical data from 52 affected individuals, we found that disruption of RARB is associated with a more variable phenotype than initially suspected, with the absence in some individuals of cardinal features of MCOPS12, such as developmental eye anomaly or motor impairment.ConclusionOur study indicates that pathogenic variants in RARB are functionally heterogeneous and associated with extensive clinical heterogeneity.

IntroductionGastric cancer (GC) is an aggressive and heterogeneous malignancy marked by cellular and molecular diversity. In GC, cancer cells invade locally in the stomach at stage I and can progress to metastasis in distant organs by stage IV, where it often becomes fatal.MethodsWe analyzed gene expression profiles from 719 stage I and stage IV GC patients across seven public datasets, conducting functional enrichment analysis to identify a gene signature linked to disease progression. Additionally, we developed an in vitro model of a simplified extracellular matrix (ECM) for cell-based assays.ResultsOur analysis identified a progression-associated gene signature (APOD, COL1A2, FSTL1, GEM, LUM, and SPARC) that characterizes stage IV GC. This signature is associated with ECM organization and epithelial-to-mesenchymal transition (EMT), both of which influence the tumor microenvironment by promoting cell invasion and triggering EMT.DiscussionThis gene signature may help identify stage I GC patients at higher risk, offering potential utility in early-stage patient management. Furthermore, our experimental ECM model may serve as a platform for investigating molecular mechanisms underlying metastatic spread in gastric cancer.

Although various tools provide proteomic information, each tool has limitations related to execution platforms, libraries, versions, and data output format. Integrating data generated from different software is a laborious process that can prolong analysis time. Here, we present FastProtein, a protein analysis pipeline that is user-friendly, easily installable, and outputs important information about subcellular location, transmembrane domains, signal peptide, molecular weight, isoelectric point, hydropathy, aromaticity, gene ontology, endoplasmic reticulum retention domains, and N-glycosylation domains. It also helps determine the presence of glycosylphosphatidylinositol and obtain functional information from InterProScan, PANTHER, Pfam, and alignment-based annotation searches. FastProtein provides the scientific community with an easy-to-use computational tool for proteomic data analysis. It is applicable to both small datasets and proteome-wide studies. It can be used through the command line interface mode or a web interface installed on a local server. FastProtein significantly enhances proteomics analysis workflows by producing multiple results in a single-step process, thereby streamlining and accelerating the overall analysis. The software is open-source and freely available. Installation and execution instructions, as well as the source code and test files generated for tool validation, are available at https://github.com/bioinformatics-ufsc/FastProtein.

Saffron (Crocus sativus L.) is one of the most expensive spices in the world due to its strong market demand combined with its labor-intensive production process, which needs a lot of labor and has significant costs. New cultivation methods and traceability systems are required to improve and valorize local Italian saffron production. In this study, we conducted a three-year trial in Umbria (Central Italy), looking for a soilless cultivation method based on wooden bins posted at a suitable height from the ground to ease the sowing of corms and harvesting of flowers. Moreover, the spice traceability could be based on investigating the genetic variability of Italian saffron populations using SNP markers. The proposed novel cultivation method showed significantly higher stigma and corm production than the traditional one. At the same time, the genetic analysis revealed a total of 55 thousand SNPs, 53 of which were specific to the Italian saffron populations suitable to start a food traceability and spice certification.

Fusarium spp., the causal agent of Fusarium wilt, cause substantial economic losses in viticulture, mainly in tropical regions. This study aimed to assess the biocontrol potential of Bacillus sp. F62 against Fusarium spp., both in vitro and in rootstock cuttings of the SO4 variety. To this end, the in vitro antagonism was evaluated through diffusible and volatile compounds synthesized by Bacillus sp. F62 on three Fusarium spp. isolates. Subsequently, the isolate FusA06-18 was selected for a rootstock cutting experiment. The vine cuttings underwent the following treatments: control, pathogen inoculation (Fus), bacterial inoculation (Bac), and bacterial followed by pathogen inoculation (Bac + Fus). Our findings revealed an average reduction of 39.1% in the mycelial growth of the pathogen through dual culture assay and a decrease of 11.6% in the Fusarium spp. radial growth due to the effects of volatile compounds. In the experiment with vine cuttings, applying Bacillus sp. F62 reduced the pathogen re-isolation frequency from 81.7% (Fus) to 63.3% (Bac + Fus). Therefore, Bacillus sp. F62 effectively suppressed the mycelial growth of Fusarium spp. and reduced the Fusarium wilt incidence in vine cuttings of the rootstock ‘SO4’.

La medición del índice de resistencia renal (IR) mediante ecografía Doppler es una técnica común para detectar problemas en el sistema vascular renal. A pesar de su aplicación previa en felinos sanos y con enfermedad renal, su utilidad clínica es incierta. Este estudio buscó correlacionar el IR con marcadores renales (creatinina, BUN/urea y densidad urinaria) en gatos adultos sanos. Se utilizaron 18 gatos mayores de 5 años de Bogotá y Zipaquirá, Colombia, en un estudio descriptivo. Se tomaron muestras de sangre periférica y se midieron BUN, urea y creatinina sérica. Se utilizó el análisis de color espectral Doppler para medir el flujo arterial renal. Los datos se analizaron con el software R® (nivel de confianza del 95%). Los valores promedio obtenidos fueron: creatinina 1,14 ± 0,3 mg/dl, BUN 32,60 ± 6,9 mg/dl, urea 64,10 ± 17,64 mg/dl, densidad urinaria 1052 ± 74,98 e IR 0,69 ± 0,071. Hubo una correlación positiva y estadísticamente significativa (P

Staphylococcus aureus is the main etiological agent of bovine mastitis worldwide and knowledge about its diversity and virulence factors is vital in controlling infections caused by this pathogen. The present study aimed to perform molecular characterization of a population of S. aureus (n=153 strains isolated from 1994 to 2014 in seven Brazilian states) by pulsed-field gel electrophoresis (PFGE) and evaluate their virulence profiles via polymerase chain reaction (PCR). PFGE identified 93 pulsotypes, with the isolates organized into 26 clusters and 20 unique pulsotypes. Predominant pulsotypes were observed, with variations according to the years of isolation and geographic origin of the isolates. Based on the PCR results for the genes encoding agglutination factors (ClfA and ClfB), binding proteins (fibronectin binding protein - FnBPA, elastin binding protein - Ebps, collagen binding protein - Cna), and toxins (Hla, Hlb and Luk-ED), 40 virulence profiles were detected. The frequency of virulence genes ranged from 58 to 98% (clfA:84.3%; clfB and hlb both 81%; hla:71.2%; fnBA:82.3%; Can:94.7%; ebps:58%; and lukED:98%). The existence of prevalent genotypes in some of the Brazilian states and the time period studied suggests that these genotypes are better adapted, with favorable characteristics in host/pathogen relationships. Genes of proven importance for S. aureus pathogenesis in bovine mastitis were widely distributed in genetically divergent populations, suggesting that most of these genes may be interesting candidates in the development of vaccines to control bovine mastitis in Brazil.

The present case report is aimed to highlight the difficulty and the reason for the delayed diagnosis of phosphaturic mesenchymal tumors, emphasizing the need of standardized protocols for diagnosis, surgery and follow-up in high-volume hospitals. The clinical signs and symptoms, diagnostic and therapeutic procedures, immunohistological features were analyzed. Delayed diagnosis of phosphaturic mesenchymal tumor was primarily due to non-specific clinical symptoms such as fatigue, muscular and bone pain, and multiple fractures. This cryptic clinical picture made the diagnosis tricky that led to treatment of patient for non-specific pain and stress fractures before to consider the tumor-induced osteomalacia syndrome. Some well-documented studies were found in the literature in which the history of trauma is a critical trigger of glomus tumors. Extra-subungual tumors most frequently occur in the knee and ankle regions, particularly among young adults, and the diagnosis is typically made approximately 7.2 years after initial symptom onset. The difficult tumor localization represented an additional obstacle to the prompt treatment, leading to delayed curative surgery.

The nonstructural protein 3 (NSP3) macrodomain of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) (Mac1) removes adenosine diphosphate (ADP) ribosylation posttranslational modifications, playing a key role in the immune evasion capabilities of the virus responsible for the coronavirus disease 2019 pandemic. Here, we determined neutron and x-ray crystal structures of the SARS-CoV-2 NSP3 macrodomain using multiple crystal forms, temperatures, and pHs, across the apo and ADP-ribose-bound states. We characterize extensive solvation in the Mac1 active site and visualize how water networks reorganize upon binding of ADP-ribose and non-native ligands, inspiring strategies for displacing waters to increase the potency of Mac1 inhibitors. Determining the precise orientations of active site water molecules and the protonation states of key catalytic site residues by neutron crystallography suggests a catalytic mechanism for coronavirus macrodomains distinct from the substrate-assisted mechanism proposed for human MacroD2. These data provoke a reevaluation of macrodomain catalytic mechanisms and will guide the optimization of Mac1 inhibitors.

This work is aimed at the experimental characterisation of air quality and thermal profile within an electric vehicle cabin, measuring at the same time the HVAC system energy consumption. Pollutant concentrations in the vehicle cabin are measured by means of a low-cost system of sensors. The effects of the HVAC system configuration, such as fresh-air and recirculation mode, on cabin air quality, are discussed. It is shown that the PM concentrations observed in recirculation mode are lower than those in fresh-air mode, while VOC concentrations are generally higher in recirculation than in fresh-air mode. The energy consumption is compared in different configurations of the HVAC system. The novelty of this work is the combined measurement of important comfort parameters such as air temperature distribution and air quality within the vehicle, together with the real time energy consumption of the HVAC system. A wider concept of comfort is enabled, based on the use of low-cost sensors in the automotive field.

The practice of serial X-ray crystallography (SX) depends on efficient, continuous delivery of hydrated protein crystals while minimizing background scattering. Of the two major types of sample delivery devices, fixed-target devices offer several advantages over widely adopted jet injectors, including: lower sample consumption, clog-free delivery, and the ability to control on-chip crystal density to improve hit rates. Here we present our development of versatile, inexpensive, and robust polymer microfluidic chips for routine and reliable room temperature serial measurements at both synchrotrons and X-ray free electron lasers (XFELs). Our design includes highly X-ray-transparent enclosing thin film layers tuned to minimize scatter background, adaptable sample flow layers tuned to match crystal size, and a large sample area compatible with both raster scanning and rotation based serial data collection. The optically transparent chips can be used both for in situ protein crystallization (to eliminate crystal handling) or crystal slurry loading, with prepared samples stable for weeks in a humidified environment and for several hours in ambient conditions. Serial oscillation crystallography, using a multi-crystal rotational data collection approach, at a microfocus synchrotron beamline (SSRL, beamline 12-1) was used to benchmark the performance of the chips. High-resolution structures (1.3-2.7 Å) were collected from five different proteins - hen egg white lysozyme, thaumatin, bovine liver catalase, concanavalin-A (type VI), and SARS-CoV-2 nonstructural protein NSP5. Overall, our modular fabrication approach enables precise control over the cross-section of materials in the X-ray beam path and facilitates chip adaption to different sample and beamline requirements for user-friendly, straightforward diffraction measurements at room temperature.