Specific Response Research Articles

Ferritin nanoparticle-based Nipah virus glycoprotein vaccines elicit potent protective immune responses in mice and hamsters

Nipah virus (NiV) is a zoonotic paramyxovirus in the genus Henipavirus that is prevalent in Southeast Asia. NiV leads to severe respiratory disease and encephalitis in humans and animals, with a mortality rate of up to 75%. Despite the grave threat to public health and global biosecurity, no medical countermeasures are available for humans. Here, based on self-assembled ferritin nanoparticles (FeNPs), we successfully constructed two candidate FeNP vaccines by loading mammalian cells expressing NiV sG (residues 71–602, FeNP-sG) and Ghead (residues 182–602, FeNP-Ghead) onto E. coli-expressed FeNPs (FeNP-sG and FeNP-Ghead, respectively) through Spycatcher/Spytag technology. Compared with sG and Ghead alone, FeNP-sG and FeNP-Ghead elicited significant NiV specific neutralizing antibody levels and T-cell responses in mice, whereas the immune response in the FeNP-sG immunized group was greater than that in the FeNP-Ghead group. These results further demonstrate that sG possesses greater antigenicity than Ghead and that FeNPs can dramatically enhance immunogenicity. Furthermore, FeNP-sG provided 100% protection against NiV challenge in a hamster model when it was administered twice at a dose of 5 μg/per animal. Our study provides not only a promising candidate vaccine against NiV, but also a theoretical foundation for the design of a NiV immunogen for the development of novel strategies against NiV infection.

Duodenal transcriptomics demonstrates signatures of tissue inflammation and immune cell infiltration in children with environmental enteric dysfunction across global centers

BackgroundEnvironmental enteric dysfunction (EED) is an inflammatory condition of the small intestine that is prevalent in children residing in low- and middle-income countries. EED is accompanied by profound histopathologic changes in the small bowel, loss of absorptive capacity, increased intestinal permeability, increased microbial translocation, and nutrient loss. ObjectivesWe sought to identify dysregulated genes and pathways that might underlie pediatric EED. MethodsRNA-sequencing libraries were generated from endoscopically obtained duodenal tissue from undernourished children with EED from 3 prospective cohorts of children with EED. The EED transcriptome was defined in comparison to North American children without EED. Weighted gene coexpression network analysis (WGCNA) was tested for gene modules associated with EED and its histologic features. ResultsThe 1784 upregulated genes in EED were highly enriched for immune and inflammatory processes, including IL-17 and JAK-STAT signaling, and cytokine–cytokine receptor interactions. The 1388 downregulated genes included genes corresponding to xenobiotic metabolism, detoxification, and antioxidant capacities. A gene coexpression module enriched for antimicrobial responses and chemokine activity was significantly associated with villous blunting, goblet cell depletion, and overall histologic severity of EED. ConclusionsThe transcriptome signatures of EED include specific innate and adaptive immune responses that are consistently elevated across study centers, coupled with reduced detoxification and antioxidant capacities. These data may have implications for targeted interventions to improve EED outcomes.

Interventions to minimise hospital winter pressures related to discharge planning and integrated care: a rapid mapping review of UK evidence.

Winter pressures are a familiar phenomenon within the National Health Service and represent the most extreme of many regular demands placed on health and social care service provision. This review focuses on a part of the pathway that is particularly problematic: the discharge process from hospital to social care and the community. Although studies of discharge are plentiful, we identified a need to focus on identifying interventions and initiatives that are a specific response to 'winter pressures'. This mapping review focuses on interventions or initiatives in relation to hospital winter pressures in the United Kingdom with either discharge planning to increase smart discharge (both a reduction in patients waiting to be discharged and patients being discharged to the most appropriate place) and/or integrated care. We conducted a mapping review of United Kingdom evidence published 2018-22. Initially, we searched MEDLINE, Health Management Information Consortium, Social Care Online, Social Sciences Citation Index and the King's Fund Library to find relevant interventions in conjunction with winter pressures. From these interventions we created a taxonomy of intervention types and a draft map. A second broader stage of searching was then undertaken for named candidate interventions on Google Scholar (Google Inc., Mountain View, CA, USA). For each taxonomy heading, we produced a table with definitions, findings from research studies, local initiatives and systematic reviews and evidence gaps. The taxonomy developed was split into structural, changing staff behaviour, changing community provision, integrated care, targeting carers, modelling and workforce planning. The last two categories were excluded from the scope. Within the different taxonomy sections we generated a total of 41 headings. These headings were further organised into the different stages of the patient pathway: hospital avoidance, alternative delivery site, facilitated discharge and cross-cutting. The evidence for each heading was summarised in tables and evidence gaps were identified. Few initiatives identified were specifically identified as a response to winter pressures. Discharge to assess and hospital at home interventions are heavily used and well supported by the evidence but other responses, while also heavily used, were based on limited evidence. There is a lack of studies considering patient, family and provider needs when developing interventions aimed at improving delayed discharge. Additionally, there is a shortage of studies that measure the longer-term impact of interventions. Hospital avoidance and discharge planning are whole-system approaches. Considering the whole health and social care system is imperative to ensure that implementing an initiative in one setting does not just move the problem to another setting. Time limitations for completing the review constrained the period available for additional searches. This may carry implications for the completeness of the evidence base identified. Further research to consider a realist review that views approaches across the different sectors within a whole system evaluation frame. This award was funded by the National Institute for Health and Care Research (NIHR) Health and Social Care Delivery Research programme (NIHR award ref: NIHR130588) and is published in full in Health and Social Care Delivery Research; Vol. 12, No. 31. See the NIHR Funding and Awards website for further award information.

Tissue specific responses to oxidative fuel source preference during heat stress in lactating dairy cows

Prolonged exposure to high environmental temperatures results in an accumulated heat load that induces a heat stress (HS) response in dairy cattle. Heat stress compromises dairy farm profitability by reducing milk yield, altering milk composition, and hindering reproductive performance. The ability to alternate between carbohydrate and lipid sources for energy production is termed metabolic flexibility (Met Flex). The objective of this study was to evaluate the Met Flex of mammary, muscle, and liver tissue in lactating dairy cows under HS and thermoneutral (TN) conditions. Sixteen Holstein cows were assigned to 1 of 2 treatment groups: pair feeding in a TN environment (PFTN) or HS. All cows experienced a 4 d TN period with ad libitum intake followed by a 4 d treatment period. Heat stress cows were exposed to a temperature humidity index (THI) ranging from 76 to 80 and the PFTN cows were exposed to a THI of 64. Milk production and health data were recorded twice daily. Semitendinosus biopsies were obtained on d 4 of each period and post-mortem mammary and liver samples were obtained on d 4 of period 2. All tissue samples were assayed for Met Flex. Activity of mitochondrial (Mit) enzymes were assessed in skeletal muscle only. Four d of HS decreased milk yield, altered milk composition, and increased respiration rate and rectal temperatures. No differences in Met Flex were observed in mammary or liver tissue during period 2. However, HS, but not PFTN conditions, lowered Met Flex of skeletal muscle by 18.3% when compared with TN ad libitum feed intake conditions of period 1. No treatment differences were observed in skeletal muscle Mit enzyme activity indicating the decrease in Met Flex occurred independently of changes in Mit function. The reduction in Met Flex of skeletal muscle during HS may contribute to reduced milk yield and warrants further investigation.

Omicron and periodontal health; a clinical exploration of gingivitis and periodontitis

Introduction: The COVID-19 pandemic has profoundly affected global health, with variants such as Omicron posing new challenges due to their increased transmissibility and potential to evade immunity. Periodontal diseases, which are highly prevalent worldwide, have been linked to various systemic conditions, including respiratory infections. Objectives: To determine how periodontal problems affect Omicron infection susceptibility, symptom severity, and outcome in individuals infected with this variety and discover oral microbiota biomarkers that indicate severe Omicron patients. Materials and Methods: To gather data, a quantitative descriptive study was instituted, utilizing a well-structured questionnaire comprising multiple-choice and Likert scale questions. The sample population, dentists in Erbil, was reached through email, followed by statistical analyses involving mean scores, standard deviations, and practices degree to interpret the data. Results: Results displayed a varied range of perceptions, with significant discrepancies highlighted through high standard deviations in specific responses, pointing to the necessity for homogenized guidelines or educational outreach. The overall mean score categorized the sentiment as ‘Medium,’ indicating a moderate level of consensus on the relationship between the Omicron variant and periodontal issues. Conclusion: This exploration unveils not only a nuanced understanding among the dentist community but also showcases domains where unanimity is deficient, calling for intensified research and educational efforts to harmonize perspectives in this pressing matter, especially amid the prevalence of the Omicron variant. The findings signify an imperative to foster aligned understandings through enhanced educational interventions in the dental community in Erbil, Kurdistan, Iraq, potentially steering towards a more cohesive approach in handling cases with pre-existing periodontal conditions amid the Omicron surge.

Interactions at the Oviposition Scar: Molecular and Metabolic Insights into Elaeagnus angustifolia's Resistance Response to Anoplophora glabripennis.

The Russian olive (Elaeagnus angustifolia), which functions as a "dead-end trap tree" for the Asian long-horned beetle (Anoplophora glabripennis) in mixed plantations, can successfully attract Asian long-horned beetles for oviposition and subsequently kill the eggs by gum. This study aimed to investigate gum secretion differences by comparing molecular and metabolic features across three conditions-an oviposition scar, a mechanical scar, and a healthy branch-using high-performance liquid chromatography and high-throughput RNA sequencing methods. Our findings indicated that the gum mass secreted by an oviposition scar was 1.65 times greater than that secreted by a mechanical scar. Significant differences in gene expression and metabolism were observed among the three comparison groups. A Kyoto Encyclopedia of Genes and Genomes annotation and enrichment analysis showed that an oviposition scar significantly affected starch and sucrose metabolism, leading to the discovery of 52 differentially expressed genes and 7 differentially accumulated metabolites. A network interaction analysis of differentially expressed metabolites and genes showed that EaSUS1, EaYfcE1, and EaPGM1 regulate sucrose, uridine diphosphate glucose, α-D-glucose-1P, and D-glucose-6P. Although the polysaccharide content in the OSs was 2.22 times higher than that in the MSs, the sucrose content was lower. The results indicated that the Asian long-horned beetle causes Russian olive sucrose degradation and D-glucose-6P formation. Therefore, we hypothesized that damage caused by the Asian long-horned beetle could enhance tree gum secretions through hydrolyzed sucrose and stimulate the Russian olive's specific immune response. Our study focused on the first pair of a dead-end trap tree and an invasive borer pest in forestry, potentially offering valuable insights into the ecological self-regulation of Asian long-horned beetle outbreaks.

Vaccinated Atlantic salmon (Salmo salar L.) maintain a specific antibody response throughout the seasonal fluctuations of a full commercial production cycle in sea: a case study

Vaccination is a critical prophylactic measure to prevent disease outbreaks in Atlantic salmon (Salmo salar L.) and multivalent oil-adjuvanted vaccines administered intraperitoneally (IP) are widely used for this purpose. The Atlantic salmon in this observational study were sampled during commercial production in a salmon farm in Northern Norway. Fish were sampled in the hatchery prior to vaccination, and further every 6 weeks after sea transfer up until slaughter. The key findings revealed that Atlantic salmon vaccinated IP with a commercial multivalent oil-based vaccine developed specific and long-lasting antibody responses against A. salmonicida, V. anguillarum, Y. ruckeri, and IPNV for up to 20 months post-vaccination. However, there was variation in the measured specific antibody response to the different antigens. Notably, the antibody responses against A. salmonicida and V. anguillarum antigens were higher and more prolonged compared to the antibody responses against Y. ruckeri and IPNV antigens. Histological examination showed presence of vaccine-induced granulomas in the pancreas at the injection site for up to 50 weeks post-vaccination and V. anguillarum antigens were found in the granulomas up to 44 weeks post-vaccination. There were also individual IgM-positive cells present in these vaccine-induced granulomas, and qPCR confirmed increased gene expression of membrane-bound and secretory IgM up to 37 weeks post-vaccination. Altogether the qPCR data of membrane and secretory IgM suggests local production of IgM in the vaccine-induced granulomas, while the head kidney might be responsible for the long-term secretion of IgM. Neither the low temperature the fish experienced for two winter seasons (below 5 °C) nor the seasonal changes in photoperiod affected the vaccine-induced specific antibody responses.

TgVax452, an epitope-based candidate vaccine targeting Toxoplasma gondii tachyzoite-specific SAG1-related sequence (SRS) proteins: immunoinformatics, structural simulations and experimental evidence-based approaches

BackgroundThe highly expressed surface antigen 1 (SAG1)-related sequence (SRS) proteins of T. gondii tachyzoites, as a widespread zoonotic parasite, are critical for host cell invasion and represent promising vaccine targets. In this study, we employed a computer-aided multi-method approach for in silico design and evaluation of TgVax452, an epitope-based candidate vaccine against T. gondii tachyzoite-specific SRS proteins.MethodsUsing immunoinformatics web-based tools, structural modeling, and static/dynamic molecular simulations, we identified and screened B- and T-cell immunodominant epitopes and predicted TgVax452’s antigenicity, stability, safety, adjuvanticity, and physico-chemical properties.ResultsThe designed protein possessed 452 residues, a MW of 44.07 kDa, an alkaline pI (6.7), good stability (33.20), solubility (0.498), and antigenicity (0.9639) with no allergenicity. Comprehensive molecular dynamic (MD) simulation analyses confirmed the stable interaction (average potential energy: 3.3799 × 106 KJ/mol) between the TLR4 agonist residues (RS09 peptide) of the TgVax452 in interaction with human TLR4, potentially activating innate immune responses. Also, a dramatic increase was observed in specific antibodies (IgM and IgG), cytokines (IFN-γ), and lymphocyte responses, based on C-ImmSim outputs. Finally, we optimized TgVax452’s codon adaptation and mRNA secondary structure for efficient expression in E. coli BL21 expression machinery.ConclusionOur findings suggest that TgVax452 is a promising candidate vaccine against T. gondii tachyzoite-specific SRS proteins and requires further experimental studies for its potential use in preclinical trials.

High-throughput transposon mutagenesis in the family Enterobacteriaceae reveals core essential genes and rapid turnover of essentiality.

The Enterobacteriaceae are a scientifically and medically important clade of bacteria, containing the model organism Escherichia coli, as well as major human pathogens including Salmonella enterica and Klebsiella pneumoniae. Essential gene sets have been determined for several members of the Enterobacteriaceae, with the Keio E. coli single-gene deletion library often regarded as a gold standard. However, it remains unclear how gene essentiality varies between related strains and species. To investigate this, we have assembled a collection of 13 sequenced high-density transposon mutant libraries from five genera within the Enterobacteriaceae. We first assess several gene essentiality prediction approaches, investigate the effects of transposon density on essentiality prediction, and identify biases in transposon insertion sequencing data. Based on these investigations, we develop a new classifier for gene essentiality. Using this new classifier, we define a core essential genome in the Enterobacteriaceae of 201 universally essential genes. Despite the presence of a large cohort of variably essential genes, we find an absence of evidence for genus-specific essential genes. A clear example of this sporadic essentiality is given by the set of genes regulating the σE extracytoplasmic stress response, which appears to have independently acquired essentiality multiple times in the Enterobacteriaceae. Finally, we compare our essential gene sets to the natural experiment of gene loss in obligate insect endosymbionts that have emerged from within the Enterobacteriaceae. This isolates a remarkably small set of genes absolutely required for survival and identifies several instances of essential stress responses masked by redundancy in free-living bacteria.IMPORTANCEThe essential genome, that is the set of genes absolutely required to sustain life, is a core concept in genetics. Essential genes in bacteria serve as drug targets, put constraints on the engineering of biological chassis for technological or industrial purposes, and are key to constructing synthetic life. Despite decades of study, relatively little is known about how gene essentiality varies across related bacteria. In this study, we have collected gene essentiality data for 13 bacteria related to the model organism Escherichia coli, including several human pathogens, and investigated the conservation of essentiality. We find that approximately a third of the genes essential in any particular strain are non-essential in another related strain. Surprisingly, we do not find evidence for essential genes unique to specific genera; rather it appears a substantial fraction of the essential genome rapidly gains or loses essentiality during evolution. This suggests that essentiality is not an immutable characteristic but depends crucially on the genomic context. We illustrate this through a comparison of our essential genes in free-living bacteria to genes conserved in 34 insect endosymbionts with naturally reduced genomes, finding several cases where genes generally regarded as being important for specific stress responses appear to have become essential in endosymbionts due to a loss of functional redundancy in the genome.

Structural and systems characterization of phosphorylation on metabolic enzymes identifies sex-specific metabolic reprogramming in obesity.

Coordination of adaptive metabolism through cellular signaling networks and metabolic response is essential for balanced flow of energy and homeostasis. Post-translational modifications such as phosphorylation offer a rapid, efficient, and dynamic mechanism to regulate metabolic networks. Although numerous phosphorylation sites have been identified on metabolic enzymes, much remains unknown about their contribution to enzyme function and systemic metabolism. In this study, we stratify phosphorylation sites on metabolic enzymes based on their location with respect to functional and dimerization domains. Our analysis reveals that the majority of published phosphosites are on oxidoreductases, with particular enrichment of phosphotyrosine (pY) sites in proximity to binding domains for substrates, cofactors, active sites, or dimer interfaces. We identify phosphosites altered in obesity using a high fat diet (HFD) induced obesity model coupled to multiomics, and interrogate the functional impact of pY on hepatic metabolism. HFD induced dysregulation of redox homeostasis and reductive metabolism at the phosphoproteome and metabolome level in a sex-specific manner, which was reversed by supplementing with the antioxidant butylated hydroxyanisole (BHA). Partial least squares regression (PLSR) analysis identified pY sites that predict HFD or BHA induced changes of redox metabolites. We characterize predictive pY sites on glutathione S-transferase pi 1 (GSTP1), isocitrate dehydrogenase 1 (IDH1), and uridine monophosphate synthase (UMPS) using CRISPRi-rescue and stable isotope tracing. Our analysis revealed that sites on GSTP1 and UMPS inhibit enzyme activity while the pY site on IDH1 induces activity to promote reductive carboxylation. Overall, our approach provides insight into the convergence points where cellular signaling fine-tunes metabolism.

Lipid Nanoparticles for Brain Tumor Theranostics: Challenges and Status.

Lipid nanoparticles have been recognized as a powerful weapon for delivering various imaging and therapeutic agents to the localized solid tumors, especially brain tumors individually or in combination. Promisingly, lipid-based nanosystems have been considered as safe delivery systems which are even approved by the Food and Drug Administration (FDA) and the European Medicines Agency (EMA). One recent spotlight of lipid nanoparticles as COVID-19 mRNA vaccines where lipid nanoparticles play an important role in effectively protecting and delivering mRNA to the desired cells. As of now, successive progress in lipid-based nanocarriers, viz., nanoliposomes, solid lipid nanoparticles, ionizable lipid nanostructures, etc., with better biochemical and biophysical stabilities, has been noticed and reported. Moreover, lipid nanostructures have been considered as versatile therapeutics platforms for a variety of diseases due to their biocompatibility, ability to protect and deliver therapeutics to the localized site, and better reproducibility and reliability. However, lipid nanoparticles still face morphological and biochemical changes upon their in vivo administration. These changes alter the specific biological and pathological response of lipid nanoparticles during their personalized brain tumor theranostics. Second, lipid nanomedicine still faces major challenges of zero premature leakage of loaded cargo, long-term colloidal stability, and off targeting. Herein, various lipid-based nanomedicines for brain tumor imaging and therapeutics "theranostics" have been reviewed and summarized considering major aspects of preclinical and clinical studies. On the other hand, engineering and biological challenges of lipid theranostics systems with relevant advantages and guidelines for clinical practice for different brain tumors have also been discussed. This review provides in-depth knowledge of lipid nanoparticle-based theranostics agents for brain tumor imaging and therapeutics.

Enhanced Particle Classification in Water Cherenkov Detectors Using Machine Learning: Modeling and Validation with Monte Carlo Simulation Datasets

The Latin American Giant Observatory (LAGO) is a ground-based extended cosmic rays observatory designed to study transient astrophysical events, the role of the atmosphere on the formation of secondary particles, and space-weather-related phenomena. With the use of a network of Water Cherenkov Detectors (WCDs), LAGO measures the secondary particle flux, a consequence of the interaction of astroparticles impinging on the atmosphere of Earth. This flux can be grouped into three distinct basic constituents: electromagnetic, muonic, and hadronic components. When a particle enters a WCD, it generates a measurable signal characterized by unique features correlating to the particle’s type and the detector’s specific response. The resulting charge histograms from these signals provide valuable insights into the flux of primary astroparticles and their key characteristics. However, these data are insufficient to effectively distinguish between the contributions of different secondary particles. In this work, we extend our previous research by using detailed simulations of the expected atmospheric response to the primary flux and the corresponding response of our WCDs to atmospheric radiation. This dataset, which was created through the combination of the outputs of the ARTI and Meiga simulation frameworks, simulated the expected WCD signals produced by the flux of secondary particles during one day at the LAGO site in Bariloche, Argentina, situated at 865 m above sea level. This was achieved by analyzing the real-time magnetospheric and local atmospheric conditions for February and March of 2012, where the resultant atmospheric secondary-particle flux was integrated into a specific Meiga application featuring a comprehensive Geant4 model of the WCD at this LAGO location. The final output was modified for effective integration into our machine-learning pipeline. With an implementation of Ordering Points to Identify the Clustering Structure (OPTICS), a density-based clustering algorithm used to identify patterns in data collected by a single WCD, we have further refined our approach to implement a method that categorizes particle groups using advanced unsupervised machine learning techniques. This allowed for the differentiation among particle types and utilized the detector’s nuanced response to each, thus pinpointing the principal contributors within each group. Our analysis has demonstrated that applying our enhanced methodology can accurately identify the originating particles with a high degree of confidence on a single-pulse basis, highlighting its precision and reliability. These promising results suggest the feasibility of future implementations of machine-leaning-based models throughout LAGO’s distributed detection network and other astroparticle observatories for semi-automated, onboard and real-time data analysis.

Cooperative thalamocortical circuit mechanism for sensory prediction errors

The brain functions as a prediction machine, utilizing an internal model of the world to anticipate sensations and the outcomes of our actions. Discrepancies between expected and actual events, referred to as prediction errors, are leveraged to update the internal model and guide our attention towards unexpected events1–10. Despite the importance of prediction-error signals for various neural computations across the brain, surprisingly little is known about the neural circuit mechanisms responsible for their implementation. Here we describe a thalamocortical disinhibitory circuit that is required for generating sensory prediction-error signals in mouse primary visual cortex (V1). We show that violating animals’ predictions by an unexpected visual stimulus preferentially boosts responses of the layer 2/3 V1 neurons that are most selective for that stimulus. Prediction errors specifically amplify the unexpected visual input, rather than representing non-specific surprise or difference signals about how the visual input deviates from the animal’s predictions. This selective amplification is implemented by a cooperative mechanism requiring thalamic input from the pulvinar and cortical vasoactive-intestinal-peptide-expressing (VIP) inhibitory interneurons. In response to prediction errors, VIP neurons inhibit a specific subpopulation of somatostatin-expressing inhibitory interneurons that gate excitatory pulvinar input to V1, resulting in specific pulvinar-driven response amplification of the most stimulus-selective neurons in V1. Therefore, the brain prioritizes unpredicted sensory information by selectively increasing the salience of unpredicted sensory features through the synergistic interaction of thalamic input and neocortical disinhibitory circuits.

Insights into Asparaginase Allergic Responses: Exploring Pharmacogenetic Influences.

Acute lymphoblastic leukemia represents the most prevalent childhood cancer. Modern chemotherapy has significantly improved outcomes, achieving EFS rates of 80% and OS rates nearing 90% in developed nations, while in developing regions, rates remain below 50%, highlighting disparities, and this difference is due to several factors. Genetic variability plays a role in these drug response disparities, presenting single-nucleotide variations (SNVs). Pharmacogenetic research aims to pinpoint these SNVs early in treatment to predict specific drug responses effectively. This review aims to explore advancements in pharmacogenetics associated with asparaginase (ASNase). ASNase plays a crucial role in the treatment of ALL and is available in three formulations: E. coli, Erwinia, and PEG ASNase. ASNase therapy presents challenges due to adverse effects, like hypersensitivity reactions. Identifying predictive markers for hypersensitivity development beforehand is crucial for optimizing treatments. Several pharmacogenetic studies have investigated the association between SNVs and the risk of hypersensitivity. Key genes include GRIA1, NFATC2, CNTO3, ARHGAP28, MYBBP1A, and HLA. Studies have highlighted associations between SNVs within these genes and hypersensitivity reactions. Notably, most pharmacogenetic investigations of hypersensitivity have focused on patients treated with E. coli, emphasizing the need for broader exploration across different formulations. Future research investigating these variants holds promise for advancing our understanding of ASNase's pharmacogenetics.

Specific Response of Bacterial Cells to β-Ionone

The biological activity and mechanisms of action of the unsaturated ketone β-ionone, a volatile compound of significantinterest for biotechnology, medicine, and agriculture, were studied. Using specific lux biosensors basedon Escherichia coli MG1655, we found that β-ionone causes oxidative stress in E. coli cells by inducing expression from the PkatG and Pdps promoters, but not from the PsoxS promoter. The effects of β-ionone on the heat shock induction (expression from the PibpA and PgrpE promoters) and on DNA damage (expression from the PcolD and PdinI promoters, SOS response) in E. coli cells were significantly weaker. β-Ionone did not cause oxidative stress in the cells of the gram-positive bacterium Bacillus subtilis.

Specific responses in soil metabolite alteration and fungal community decline to the long-term monocropping of lisianthus

Continuous cropping obstacle (CCO) is becoming a serious threat to the quality production of lisianthus (Eustoma grandiflorum). However, investigations of the causal mechanism of CCO in lisianthus are still lacking. To this end, soil biotic dynamics and metabolites alterations were illuminated using metagenomic sequencing and metabolomic profiling along a chronosequence that has cultivated lisianthus from 2 weeks to 6 years. Soil EC and nutrient contents increased significantly as the cultivation duration prolonged. Fungal community has undergone significant decline in their diversity and biomass. Nevertheless, no apparent accumulation of pathogenicity-related microbes was observed along the investigated chronosequence. The alterations of soil metabolites along the chronosequence reflect the decline of microbial activity, the accumulation of phytotoxic compounds and the loss of metabolite involved in beneficial plant-microbe interactions. As a result, lisianthus plants in the longer continuous cropping soil experienced stressful condition as evidenced by the increasing trend of stress responsive metabolites and the overall soil function deterioration as indicated by soil enzymes. Hierarchical partitioning analysis showed that the investigated factors can explain a total of 92.25 % of the variations in the soil function degradation. Of which, soil metabolite alteration was the most important factor in explaining the CCO development, covering 24.55 % of the explainable variations. Spearman's correlation results further implied that soil metabolite alteration is the main driver in the development of CCO in lisianthus, especially by mediating the plant-microbes interactions. Our findings revealed a general pattern of soil biotic and abiotic response to the continuous monocropping of lisianthus and suggest that the overcome of the CCO in lisianthus requires further studies focusing on the crucial metabolites and their interactions with microbes.

Effects of plant health protecting product applications on the root-associated microbiota of apple saplings and strawberries

The root-associated microbiome has been of keen research interest due to its large potential for increasing plant performance. In agricultural systems, the use of plant health protecting products (PHPP) is essential for obtaining high yields and quality. Knowledge about the impact of PHPP applications on the root-associated microbiome remains limited, especially when applied above ground. We hypothesized that PHPP application leads to specific plant-mediated responses in the rhizosphere microbiota. We applied different PHPPs on greenhouse-grown apple and strawberry plants, cultivated in different soils. The effects of above ground PHPP applications on the root-associated microbiota were analyzed by 16S rRNA gene amplicon sequencing. No consistent effects were observed across three experimental trials, though alpha diversity tended to be decreased after PHPP application. In some cases, shifts in community composition became evident (PERMANOVA R2 = 0.192, p = 0.001) or increases in dispersion (PERMDISP Padj = 0.040). Movento® treatments caused in part deterministic shifts in community assembly, whereas Serenade®ASO treatments caused rather stochastic shifts. Effects were concluded to be probably transient and to disappear without continuous product application. These rather heterogenous temporary responses are in line with the concept of the ‘Anna Karenina Principle’, under which terms PHPP applications can act as mild stress factors and slightly and transiently alter the below ground bacterial community. These findings indicate that above ground PHPP applications can have product specific implications for the root-associated microbiome, which should be considered in the context of microbiome management strategies.

Developing and validating a discrete-event simulation model of multiple myeloma disease outcomes and treatment pathways using a national clinical registry.

Multiple myeloma is a haematological malignancy typically characterised by neoplastic plasma cell infiltration of the bone marrow. Treatment for multiple myeloma consists of multi-line chemotherapy with or without autologous stem cell transplantation and has been rapidly evolving in recent years. However, clinical trials are unable to provide patients and clinicians with long-term prognostic information nor policymakers with the full body of evidence needed to perform economic evaluation of new therapies or make reimbursement decisions. To address these limitations of the available evidence, this study aimed to develop and validate the EpiMAP Myeloma model, a discrete-event simulation model of multiple myeloma disease outcomes and treatment pathways. Risk equations were estimated using the Australian and New Zealand Myeloma & Related Diseases Registry after multiple imputation of missing data. Risk equation coefficients were combined with multiple myeloma patients at diagnosis from the Registry to perform the simulation. The model was validated with 100 bootstraps of an out-of-sample prediction analysis using a 70/30 split of the 4,121 registry patients diagnosed between 2009 and 2023, resulting in 2,884 and 1,237 patients in the training and validation cohorts, respectively. For 90% of the 120 months in the 10-year post-diagnosis period, there was no significant difference in overall survival between the validation and simulated cohorts. These results highlight that the EpiMAP Myeloma model is robust at predicting multiple myeloma disease outcomes and treatment pathways in Australia & New Zealand. In the future, clinicians will be able to use the EpiMAP Myeloma model to provide personalised estimates of life expectancy to patients based on their specific characteristics, disease stage, and response to treatment. Policymakers will also be able to use the model to perform economic evaluation, to forecast the number of patients receiving treatment at different stages, and to determine the downstream impact of listing new, effective therapies.

Lung megakaryocytes engulf inhaled airborne particles to promote intrapulmonary inflammation and extrapulmonary distribution

Many lung immune cells are known to respond to inhaled particulate matter. However, current known responses cannot explain how particles induce thrombosis in the lung and how they translocate to distant organs. Here, we demonstrate that lung megakaryocytes (MKs) in the alveolar and interstitial regions display location-determined characteristics and act as crucial responders to inhaled particles. They move rapidly to engulf particles and become activated with upregulation in inflammatory responses and thrombopoiesis. Comprehensive in vivo, in vitro and ex vivo results unraveled that MKs were involved in particle-induced lung damages and shed particle-containing platelets into blood circulation. Moreover, MK-derived platelets exhibited faster clotting, stronger adhesion than normal resting platelets, and inherited the engulfed particles from parent MKs to assist in extrapulmonary particle transportation. Our findings collectively highlight that the specific responses of MKs towards inhaled particles and their roles in facilitating the translocation of particles from the lungs to extrapulmonary organs for clearance.

Characterization of the Monkeypox Virus [MPX]-Specific Immune Response in MPX-Cured Individuals Using Whole Blood to Monitor Memory Response.

Monkeypox (Mpox) is a zoonotic disease caused by monkeypox virus (MPXV), an Orthopoxvirus (OPXV). Since we are observing the first MPXV outbreak outside the African continent, the general population probably does not have a pre-existing memory response for MPXV but may have immunity against the previous smallpox vaccine based on a live replicating Vaccinia strain (VACV). Using a whole blood platform, we aim to study the MPXV- T-cell-specific response in Mpox-cured subjects. We enrolled 16 subjects diagnosed with Mpox in the previous 3-7 months and 15 healthy donors (HD) with no recent vaccination history. Whole blood was stimulated overnight with MPXV and VACV peptides to elicit CD4 and CD8 T-cell-specific responses, which were evaluated by ELISA and multiplex assay. Mpox-cured subjects showed a significant IFN-γ T-cell response to MPXV and VACV. Besides IFN-γ, IL-6, IP-10, IL-8, IL-2, G-CSF, MCP-1, MIP1-α, MIP-1β, IL-1Rα, and IL-5 were significantly induced after specific stimulation compared to the unstimulated control. The specific response was mainly induced by the CD4 peptides MPX-CD4-E and VACV-CD4. We showed that MPXV-specific responses have a mixed Th1- and Th2-response in a whole blood platform assay, which may be useful for monitoring the specific immunity induced by vaccination or infection.