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Journal of Biomedical Science Jun 2024Enterovirus 71 (EV-A71) causes Hand, Foot and Mouth Disease (HFMD) in children and has been associated with neurological complications. The molecular mechanisms involved...
BACKGROUND
Enterovirus 71 (EV-A71) causes Hand, Foot and Mouth Disease (HFMD) in children and has been associated with neurological complications. The molecular mechanisms involved in EV-A71 pathogenesis have remained elusive.
METHODS
A siRNA screen in EV-A71 infected-motor neurons was performed targeting 112 genes involved in intracellular membrane trafficking, followed by validation of the top four hits using deconvoluted siRNA. Downstream approaches including viral entry by-pass, intracellular viral genome quantification by qPCR, Western blot analyses, and Luciferase reporter assays allowed determine the stage of the infection cycle the top candidate, RAB11A was involved in. Proximity ligation assay, co-immunoprecipitation and multiplex confocal imaging were employed to study interactions between viral components and RAB11A. Dominant negative and constitutively active RAB11A constructs were used to determine the importance of the protein's GTPase activity during EV-A71 infection. Mass spectrometry and protein interaction analyses were employed for the identification of RAB11A's host interacting partners during infection.
RESULTS
Small GTPase RAB11A was identified as a novel pro-viral host factor during EV-A71 infection. RAB11A and RAB11B isoforms were interchangeably exploited by strains from major EV-A71 genogroups and by Coxsackievirus A16, another major causative agent of HFMD. We showed that RAB11A was not involved in viral entry, IRES-mediated protein translation, viral genome replication, and virus exit. RAB11A co-localized with replication organelles where it interacted with structural and non-structural viral components. Over-expression of dominant negative (S25N; GDP-bound) and constitutively active (Q70L; GTP-bound) RAB11A mutants had no effect on EV-A71 infection outcome, ruling out RAB11A's involvement in intracellular trafficking of viral or host components. Instead, decreased ratio of intracellular mature viral particles to viral RNA copies and increased VP0:VP2 ratio in siRAB11-treated cells supported a role in provirion maturation hallmarked by VP0 cleavage into VP2 and VP4. Finally, chaperones, not trafficking and transporter proteins, were found to be RAB11A's top interacting partners during EV-A71 infection. Among which, CCT8 subunit from the chaperone complex TRiC/CCT was further validated and shown to interact with viral structural proteins specifically, representing yet another novel pro-viral host factor during EV-A71 infection.
CONCLUSIONS
This study describes a novel, unconventional role for RAB11A during viral infection where it participates in the complex process of virus morphogenesis by recruiting essential chaperone proteins.
Topics: rab GTP-Binding Proteins; Enterovirus A, Human; Humans; Molecular Chaperones; Virus Replication
PubMed: 38943128
DOI: 10.1186/s12929-024-01053-2 -
BMC Infectious Diseases Jun 2024Intravenous fosfomycin (IVFOF) is gaining interest in severe infections. Its use may be limited by adverse events (AEs). Little experience exists on IVFOF therapeutic...
BACKGROUND
Intravenous fosfomycin (IVFOF) is gaining interest in severe infections. Its use may be limited by adverse events (AEs). Little experience exists on IVFOF therapeutic drug monitoring (TDM) in real-life setting.
PATIENTS AND METHODS
Retrospective study of patients receiving IVFOF for > 48 h at Policlinico Hospital (Milan, Italy) from 01/01/2019 to 01/01/2023. AEs associated to IVFOF graded CTCAE ≥ II were considered. Demographic and clinical risk factors for IVFOF-related AEs were analysed with simple and multivariable regression models. The determination of IVFOF TDM was made by a rapid ultraperformance liquid chromatography mass spectrometry method (LC-MS/MS) on plasma samples. The performance of TDM (trough levels (Cmin) in intermittent infusion, steady state levels (Css) in continuous infusion) in predicting AEs ≤ 5 days after its assessment was evaluated.
RESULTS
Two hundred and twenty-four patients were included. At IVFOF initiation, 81/224 (36.2%) patients were in ICU and 35/224 (15.7%) had septic shock. The most frequent infection site was the low respiratory tract (124/224, 55.4%). Ninety-five patients (42.4%) experienced ≥ 1AEs, with median time of 4.0 (2.0-7.0) days from IVFOF initiation. Hypernatremia was the most frequent AE (53/224, 23.7%). Therapy discontinuation due to AEs occurred in 38/224 (17.0%). ICU setting, low respiratory tract infections and septic shock resulted associated with AEs (RR 1.59 (95%CI:1.09-2.31), 1.46 (95%CI:1.03-2.07) and 1.73 (95%CI:1.27-2.37), respectively), while IVFOF daily dose did not. Of the 68 patients undergone IVFOF TDM, TDM values predicted overall AEs and hypernatremia with AUROC of 0.65 (95%CI:0.44-0.86) and 0.91 (95%CI:0.79-1.0) respectively for Cmin, 0.67 (95%CI:0.39-0.95) and 0.76 (95%CI:0.52-1.0) respectively for Css.
CONCLUSIONS
We provided real world data on the use of IVFOF-based regimens and associated AEs. IVFOF TDM deserves further research as it may represent a valid tool to predict AEs.
KEY POINTS
Real world data on intravenous fosfomycin for severe bacterial infections. AEs occurred in over 40% (therapy discontinuation in 17%) and were related to baseline clinical severity but not to fosfomycin dose. TDM showed promising results in predicting AEs.
Topics: Humans; Fosfomycin; Female; Male; Retrospective Studies; Anti-Bacterial Agents; Middle Aged; Drug Monitoring; Risk Factors; Aged; Administration, Intravenous; Italy; Adult; Tandem Mass Spectrometry
PubMed: 38943088
DOI: 10.1186/s12879-024-09541-4 -
BMC Medical Informatics and Decision... Jun 2024Clinical medicine offers a promising arena for applying Machine Learning (ML) models. However, despite numerous studies employing ML in medical data analysis, only a... (Review)
Review
BACKGROUND
Clinical medicine offers a promising arena for applying Machine Learning (ML) models. However, despite numerous studies employing ML in medical data analysis, only a fraction have impacted clinical care. This article underscores the importance of utilising ML in medical data analysis, recognising that ML alone may not adequately capture the full complexity of clinical data, thereby advocating for the integration of medical domain knowledge in ML.
METHODS
The study conducts a comprehensive review of prior efforts in integrating medical knowledge into ML and maps these integration strategies onto the phases of the ML pipeline, encompassing data pre-processing, feature engineering, model training, and output evaluation. The study further explores the significance and impact of such integration through a case study on diabetes prediction. Here, clinical knowledge, encompassing rules, causal networks, intervals, and formulas, is integrated at each stage of the ML pipeline, resulting in a spectrum of integrated models.
RESULTS
The findings highlight the benefits of integration in terms of accuracy, interpretability, data efficiency, and adherence to clinical guidelines. In several cases, integrated models outperformed purely data-driven approaches, underscoring the potential for domain knowledge to enhance ML models through improved generalisation. In other cases, the integration was instrumental in enhancing model interpretability and ensuring conformity with established clinical guidelines. Notably, knowledge integration also proved effective in maintaining performance under limited data scenarios.
CONCLUSIONS
By illustrating various integration strategies through a clinical case study, this work provides guidance to inspire and facilitate future integration efforts. Furthermore, the study identifies the need to refine domain knowledge representation and fine-tune its contribution to the ML model as the two main challenges to integration and aims to stimulate further research in this direction.
Topics: Machine Learning; Humans; Decision Support Systems, Clinical
PubMed: 38943085
DOI: 10.1186/s12911-024-02582-4 -
BMC Neurology Jun 2024Neuropsychological symptoms in the Cognitive, Energetic, Behavioural, and Affective (CEBA) domains are common in people with multiple sclerosis (PwMS) and can negatively... (Observational Study)
Observational Study
Protocol for the MS-CEBA study: an observational, prospective cohort study identifying Cognitive, Energetic, Behavioural and Affective (CEBA) profiles in Multiple Sclerosis to guide neuropsychological treatment choice.
BACKGROUND
Neuropsychological symptoms in the Cognitive, Energetic, Behavioural, and Affective (CEBA) domains are common in people with multiple sclerosis (PwMS) and can negatively affect societal participation. The current study aims to investigate whether there are combinations of symptoms in the different CEBA domains that consistently occur together, that is, if there are CEBA profiles that can be identified. If so, this study aims to develop a screening instrument identifying CEBA profiles in PwMS to select the most suitable neuropsychological rehabilitation treatment for a given CEBA profile and consequently improve the societal participation of PwMS.
METHODS
This study is an observational, prospective cohort study consisting of 3 phases. Phase 1 focuses on the identification of CEBA profiles in a large sample of PwMS (n = 300). Phase 2 focuses on validating these CEBA profiles through replication of results in a new sample (n = 100) and on the development of the screening instrument. Phase 3 focuses on qualitatively evaluating in a small group of PwMS whether the selected treatment is suitable for the given CEBA profile or whether existing neuropsychological treatments should be adapted to meet the needs of PwMS suffering from symptoms in multiple CEBA domains simultaneously. Primary outcome is the CEBA profile, which will be derived from performance on neuropsychological assessment consisting of tests and questionnaires regarding the CEBA domains using a latent profile analysis. Inclusion criteria include MS diagnosis, sufficient ability in the Dutch language, and an age between 18 and 70 years.
DISCUSSION
The results of the current study will contribute to a more comprehensive understanding of the entire spectrum of neuropsychological symptoms in PwMS. Identification of possible CEBA profiles, and accordingly, the development of a screening instrument determining the CEBA profile of PwMS in clinical practice, contributes to the timely referral of PwMS to the most suitable neuropsychological rehabilitation treatment. If necessary, adjustments to existing treatments will be suggested in order to sufficiently meet the needs of PwMS. All of this with the ultimate aim to improve societal participation, and thereby quality of life, of PwMS.
TRIAL REGISTRATION
Dutch Central Committee on Research Involving Human Subjects (CCMO) NL83954.042.23; ClinicalTrials.gov NCT06016309.
Topics: Humans; Multiple Sclerosis; Prospective Studies; Neuropsychological Tests; Male; Female; Adult; Middle Aged; Cohort Studies
PubMed: 38943063
DOI: 10.1186/s12883-024-03737-6 -
Communications Chemistry Jun 2024Glycosylated derivatives of natural product polyphenols display a spectrum of biological activities, rendering them critical for both nutritional and pharmacological...
Glycosylated derivatives of natural product polyphenols display a spectrum of biological activities, rendering them critical for both nutritional and pharmacological applications. Their enzymatic synthesis by glycosyltransferases is frequently constrained by the limited repertoire of characterized enzyme-catalyzed transformations. Here, we explore the glycosylation capabilities and substrate preferences of newly identified plant uridine diphosphate (UDP)-dependent glycosyltransferases (UGTs) within the UGT72 and UGT84 families, with particular focus on natural polyphenol glycosylation from UDP-glucose. Four UGTs are classified according to their phylogenetic relationships and reaction products, identifying them as biocatalysts for either glucoside (UGT72 enzymes) or glucose ester (UGT84 members) formation from selected phenylpropanoid compounds. Detailed kinetic evaluations expose the unique attributes of these enzymes, including their specific activities and regio-selectivities towards diverse polyphenolic substrates, with product characterizations validating the capacity of UGT84 family members to perform di-O-glycosylation on flavones. Sequence analysis coupled with structural predictions through AlphaFold reveal an unexpected absence of a conserved threonine residue across all four enzymes, a trait previously linked to pentosyltransferases. This comparative analysis broadens the understood substrate specificity range for UGT72 and UGT84 enzymes, enhancing our understanding of their utility in the production of natural phenolic glycosides. The findings from this in-depth characterization provide valuable insights into the functional versatility of UGT-mediated reactions.
PubMed: 38942997
DOI: 10.1038/s42004-024-01231-1 -
Scientific Reports Jun 2024Understanding the cellular and molecular mechanisms of inflammation requires robust animal models. Sheep are commonly used in immune-related studies, yet the validity of... (Comparative Study)
Comparative Study
Understanding the cellular and molecular mechanisms of inflammation requires robust animal models. Sheep are commonly used in immune-related studies, yet the validity of sheep as animal models for immune and inflammatory diseases remains to be established. This cross-species comparative study analyzed the in vitro inflammatory response of ovine (oPBMCs) and human PBMCs (hPBMCs) using mass spectrometry, profiling the proteome of the secretome and whole cell lysate. Of the entire cell lysate proteome (oPBMCs: 4217, hPBMCs: 4574 proteins) 47.8% and in the secretome proteome (oPBMCs: 1913, hPBMCs: 1375 proteins) 32.8% were orthologous between species, among them 32 orthologous CD antigens, indicating the presence of six immune cell subsets. Following inflammatory stimulation, 71 proteins in oPBMCs and 176 in hPBMCs showed differential abundance, with only 7 overlapping. Network and Gene Ontology analyses identified 16 shared inflammatory-related terms and 17 canonical pathways with similar activation/inhibition patterns in both species, demonstrating significant conservation in specific immune and inflammatory responses. However, ovine PMBCs also contained a unique WC1γδ T-cell subset, not detected in hPBMCs. Furthermore, differences in the activation/inhibition trends of seven canonical pathways and the sets of DAPs between sheep and humans, emphasize the need to consider interspecies differences in translational studies and inflammation research.
Topics: Humans; Animals; Sheep; Leukocytes, Mononuclear; Proteomics; Inflammation; Proteome
PubMed: 38942936
DOI: 10.1038/s41598-024-66059-0 -
Scientific Reports Jun 2024Viscum album L. (VA) is a unique plant with regard to its biological content. It is rich in many different metabolites with high potential in various spheres of human...
Viscum album L. (VA) is a unique plant with regard to its biological content. It is rich in many different metabolites with high potential in various spheres of human activity. We conducted a pilot study with 5 VA aqueous extracts of different host-tree species for pre-sowing treatment of Cucurbita maxima 'Hokkaido orange' seeds. We set the following objectives consisting of hypotheses (1) H01 is based on different effects of tested VA extracts depending on host trees and time of pre-treatment; (2) H02 focuses on the allopathic properties of the tested extracts affecting the plant growth and development by dose-response relationship; (3) A01 considers highly biologically active compounds of VA extracts also containing allelochemicals that can be used to regulate plant growth processes and create eco-friendly and resilient cities. The analysis of the stimulatory allelopathy index for 7 parameters demonstrates the direct effect of VA extracts in 62.3% of cases. The variability of the broad spectrum of effects of VA extracts of different host trees on the ontogenesis of C. maxima plants shows the presence of potential allelochemicals, resulting from the vital products of the host-parasite relationship. These effects are not fully explained by total polyphenol content and antioxidant activity as in previous studies of other mistletoe species. The authors consider this work a pilot study that expands the areas of application of VA extracts and knowledge about potential sources of allelochemicals.
Topics: Cucurbita; Plant Extracts; Viscum album; Allelopathy; Pilot Projects; Pheromones; Seeds
PubMed: 38942921
DOI: 10.1038/s41598-024-65918-0 -
Scientific Reports Jun 2024The increasing water contamination by toxic heavy metals, particularly hexavalent chromium, has become a significant environmental concern. This study explores the...
The increasing water contamination by toxic heavy metals, particularly hexavalent chromium, has become a significant environmental concern. This study explores the pyrolysis of termite-processed biomass, specifically Pinus elliottii particleboard and its termite droppings (TDs), to produce biochar and its application for chromium (VI) adsorption. Termite droppings, rich in lignin, and particleboard, rich in cellulose, were pyrolyzed at various temperatures to assess the effect of biomass composition on biochar properties. The study found that lignin-rich termite droppings produced biochar with higher fixed carbon content and specific surface area than cellulose-rich particleboard biochar. FTIR and Raman spectroscopy revealed significant molecular structure changes during pyrolysis, which influenced the adsorption capabilities of the biochar. Adsorption experiments demonstrated that TD biochar exhibited significantly higher chromium (VI) adsorption capacity, attributed to its distinct chemical composition and enhanced surface properties due to higher lignin content. These findings underscore the crucial role of lignin in producing efficient biochar for heavy metal adsorption, highlighting the practical applicability of termite-processed biomass in water purification technologies.
Topics: Charcoal; Lignin; Chromium; Animals; Cellulose; Biomass; Adsorption; Isoptera; Water Pollutants, Chemical; Pyrolysis; Pinus; Water Purification; Spectroscopy, Fourier Transform Infrared
PubMed: 38942919
DOI: 10.1038/s41598-024-65959-5 -
Scientific Reports Jun 2024Particles containing alpha (α) nuclides were identified from sediment in stagnant water in the Unit 3 reactor building of the Fukushima Daiichi Nuclear Power Station...
Particles containing alpha (α) nuclides were identified from sediment in stagnant water in the Unit 3 reactor building of the Fukushima Daiichi Nuclear Power Station (FDiNPS). We analyzed different concentrations of α-nuclide samples collected at two sampling sites, the torus room and the main steam isolation valve (MSIV) room. The solids in the stagnant water samples were classified, and the uranium (U) and total alpha concentrations of each fraction were measured by dissolution followed by inductively coupled plasma mass spectrometry and α-spectrometry. Most of the α-nuclides in the stagnant water samples from the torus and MSIV rooms were in particle fractions larger than 10 μm. We detected uranium-bearing particles ranging from sub-µm to 10 µm in size by scanning electron microscopy-energy-dispersive X-ray (SEM-EDX) observations. The chemical forms of U particles were determined in U-Zr oxides, oxidized UO, and UO with micro-Raman spectroscopy. Other short-lived α-nuclides (plutonium [Pu], americium [Am], and curium [Cm]) were detected by alpha track detection, and the particles with α-nuclides was characterized by SEM-EDX analysis. α-nuclide-containing particles with several tens to several 100 µm in size mainly comprised iron (Fe) oxyhydroxides. In addition, we detected adsorbed U onto Fe oxyhydroxide particles in the MSIV room sample, which indicated nuclear fuel dissolution and secondary U accumulation. This study clarifies the major characteristics of U and other α-nuclides in sediment in stagnant water in the FDiNPS Unit 3 reactor building, which significantly contribute to the consideration of removal methods for particles containing α-nuclides in the stagnant water.
PubMed: 38942918
DOI: 10.1038/s41598-024-65522-2 -
Scientific Reports Jun 2024The tobacco alkaloid nicotine is known for its activation of neuronal nicotinic acetylcholine receptors. Nicotine is consumed in different ways such as through...
The tobacco alkaloid nicotine is known for its activation of neuronal nicotinic acetylcholine receptors. Nicotine is consumed in different ways such as through conventional smoking, e-cigarettes, snuff or nicotine pouches. The use of snuff has been associated with several adverse health effects, such as inflammatory reactions of the oral mucosa and oral cavity cancer. We performed a metabolomic analysis of nicotine-exposed THP-1 human monocytes. Cells were exposed to 5 mM of the alkaloid for up to 4 h, and cell extracts and medium subjected to untargeted liquid chromatography high-resolution mass spectrometry. Raw data processing revealed 17 nicotine biotransformation products. Among these, cotinine and nornicotine were identified as the two major cellular biotransformation products. The application of multi- and univariate statistical analyses resulted in the annotation, up to a certain level of identification, of 12 compounds in the cell extracts and 13 compounds in the medium that were altered by nicotine exposure. Of these, four were verified as methylthioadenosine, cytosine, uric acid, and L-glutamate. Methylthioadenosine levels were affected in both cells and the medium, while cytosine, uric acid, and L-glutamate levels were affected in the medium only. The effects of smoking on the pathways involving these metabolites have been previously demonstrated in humans. Most of the other discriminating compounds, which were merely tentatively or not fully identified, were amino acids or amino acid derivatives. In conclusion, our preliminary data suggest that some of the potentially adverse effects related to smoking may also be expected when nicotine is consumed via snuff or nicotine pouches.
Topics: Humans; Nicotine; Metabolomics; Monocytes; Mass Spectrometry; THP-1 Cells; Cotinine; Chromatography, Liquid; Metabolome; Glutamic Acid
PubMed: 38942832
DOI: 10.1038/s41598-024-65733-7