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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.

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.

Abstract Exosomes mediate cell-to-cell crosstalk involving a variety of biomolecules through an intricate signaling network. In recent years, the pivotal role of exosomes and their non-coding RNAs cargo in the development and progression of several cancer types clearly emerged. In particular, tumor bulk and its microenvironment co-evolve through cellular communications where these nanosized extracellular vesicles are among the most relevant actors. Knowledge about the cellular, and molecular mechanisms involved in these communications will pave the way for novel exosome-based delivery of therapeutic RNAs as well as innovative prognostic/diagnostic tools. Despite the valuable therapeutic potential and clinical relevance of exosomes, their role on sarcoma has been vaguely reported because the rarity and high heterogeneity of this type of cancer. Here, we dissected the scientific literature to unravel the multifaceted role of exosomal non-coding RNAs as mediator of cell-to-cell communications in the sarcoma subtypes.

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.

Abstract “The Power of Museums” was the General Conference on the central theme of the International Council of Museums (ICOM) in 2022 (Prague). This article aims to discuss the ability of ethnographic collections to combat prejudice and promote freedom, cultural diversity, religious diversity, and democracy, among other aspects of human rights. Here, we propose a theoretical reflection on the power of museums, considering the context of the decolonization of ethnographic collections in Brazil. To illustrate the discussion, we briefly present two recent cases in the country that refer to the appreciation of indigenous, religious, and cultural diversity. The first is related to a photographic collection of indigenous peoples that is part of the Museu Paranaense (MUPA) collection in Curitiba (southern Brazil). The second case involves collections from a group of people who descend from enslaved people and members of Terreiros de Umbanda and Candomblé in the city of Rio de Janeiro (southeastern region), which is now preserved at the Museu da Republica. For this purpose, we present a brief context on the historical development of Brazil, its contemporary museum universe, and Associação Brasileira de Antropologia’s initiative to map ethnographic collections in Brazil. Both cases reveal experiences of decolonizing ethnographic collections with the direct involvement of the associated communities. These cases demonstrate how it is possible to deconstruct collections shaped within colonialist and prejudiced perspectives by presenting new narratives that appreciate cultures that were previously.

Abstract Among marine ecosystems globally, those in the Mediterranean Sea, are facing many threats. New technologies are crucial for enhancing our understanding of marine habitats and ecosystems, which can be complex and resource-intensive to analyse using traditional techniques. We tested, for the first time, an integrated multi-platform approach for mapping the coastal benthic habitat in the Civitavecchia (northern Latium, Italy) coastal area. This approach includes the use of an Unmanned Surface Vehicle (USV), a Remote Operated Vehicle (ROV), and in situ measurements of ecosystem functionality. The echosounder data allowed us to reconstruct the distribution of bottom types, as well as the canopy height and coverage of the seagrass Posidonia oceanica. Our study further involved assessing the respiration (Rd) and net primary production (NCP) rates of P. oceanica and its associated community through in situ benthic chamber incubation. By combining these findings with the results of USV surveys, we were able to develop a preliminary spatial distribution model for P. oceanica primary production (PP-SDM). The P. oceanica PP-SDM was applied between the depths of 8 and 10 m in the studied area and the obtained results showed similarities with other sites in the Mediterranean Sea. Though in the early stages, our results highlight the significance of multi-platform observation data for a thorough exploration of marine ecosystems, emphasizing their utility in forecasting biogeochemical processes in the marine environment.

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

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’.

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.

ABSTRACT Objective Patients with cancer often undergo multiple extended treatments that decrease their quality of life. However, the quality of life of women with breast cancer after they undergo treatment remains underexplored in Brazil. Therefore, this study determined sociodemographic, behavioral, and clinical factors related to the post-treatment quality of life of women with breast cancer. Methods This cross-sectional study involved 101 women diagnosed with breast cancer between 2014 and 2016 and treated at a Brazilian Oncology Reference Service. Data were collected from them using face-to-face surveys. Quality of life was evaluated using the European Organization for the Research and Treatment of Cancer Core Quality of Life questionnaire (EORTC QLQ-C30) and EORTC Breast Cancer-specific Quality of Life questionnaire (EORTC QLQ-BR23). The data collected were analyzed using Student’s t-test and Mann-Whitney U test. Results The median score on the global health, functional, and symptom scales of the EORTC QLQ-C30 was 75.00 (Interquartile range=33.33), 75.99 (Standard deviation [SD]=19.26), and 19.67 (SD=16.91), respectively. The mean score on the functional and symptom scales of the EORTC QLQ-BR23 was 61.89 (SD=17.21) and 20.12 (SD=16.94), respectively. Furthermore, higher post-treatment quality of life was found to be associated with being aged 50 or more, being Black, having eight or more years of education, having a partner, having a paying job, receiving treatment from the private healthcare system, having a higher income, living in the municipality where healthcare services are availed, engaging in physical activity, not smoking, being more religious, having more social support, not being overweight, having no comorbidities, and undergoing lumpectomy. Conclusion Sociodemographic, behavioral, and clinical factors significantly impact the quality of life of women who undergo breast cancer treatment. Implementing interventions that improve health and reducing inequalities in the access to healthcare services can improve the quality of life of these patients.

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 study aimed to evaluate the addition of liquid cellulose, produced by Humicola grisea, in 21-day-old broiler chickens’ diets. The treatments comprised control rations of corn and soybean meal and rations to which 500 mL/t and 1000 mL/t of cellulase were added. A total of 180 male broiler chickens were used, distributed in a completely randomized design, with three treatments and six replicates. Broiler chicken performance was monitored during the period from 1 to 21 days old. Significant effects were detected for digestibility only between four and seven days old, when a reduced dry matter nitrogen intake was recorded, and for nitrogen digestibility in the broilers fed cellulase-supplemented rations at a dose of 1000 m/L. Among the analyzed digestive organs, only the biometrics of the large intestine were affected significantly at seven days old. The absolute weights of the liver and pancreas and the activities of amylase, alkaline phosphatase, and transaminases were not affected significantly, indicating that cellulase did not affect the metabolism of these organs. No significant effect was detected in the serum for electrolytes, total protein, or alkaline phosphatase. So, the addition of liquid cellulase produced by Humicola grisea did not affect performance and metabolism in 21-day-old broiler chickens.

Abstract Earth’s atmosphere, whose ionization stability plays a fundamental role for the evolution and endurance of life, is exposed to the effect of cosmic explosions producing high energy Gamma-ray-bursts. Being able to abruptly increase the atmospheric ionization, they might deplete stratospheric ozone on a global scale. During the last decades, an average of more than one Gamma-ray-burst per day were recorded. Nevertheless, measurable effects on the ionosphere were rarely observed, in any case on its bottom-side (from about 60 km up to about 350 km of altitude). Here, we report evidence of an intense top-side (about 500 km) ionospheric perturbation induced by significant sudden ionospheric disturbance, and a large variation of the ionospheric electric field at 500 km, which are both correlated with the October 9, 2022 Gamma-ray-burst (GRB221009A).

Abstract We report the molecular basis of Aspergillus fumigatus oryzin, allergen Asp f 13, or alkaline proteinase ALP1, containing the sequence motif His–Asp–Ser of the subtilisin family, structure, and function at atomic detail. Given the resolution of the data (1.06 Å), we use fragment molecular replacement with ideal polyalanine α-helices to determine the first crystal structure of oryzin. We probe the catalytic serine through formation of an irreversible bond to a small molecule compound, specifically labeling it, describing the amino acid residues performing the catalytic function. Defined by a self-processed pro-peptide, the active site architecture shapes up pocket-like subsites that bind to and unveil the S1′–S4′ substrate binding preferences. We use molecular modeling to dock a model of the pro-peptide in the S1–S4 region and to dock collagen along the active site cleft. Opposite to the face harboring the catalytic serine, the enzyme binds to a calcium ion in a binding site created by backbone flipping. We use thermal unfolding to show that this metal ion provides structural stability. With no known host inhibitor identified thus far, this structure may hasten the progress of developing new therapeutic agents for diseases caused by pathogenic fungi.