Significant Induction Research Articles

P0075 Evaluation of the modulatory potential of polyphenolic compounds on intestinal inflammation at the transcriptomic level

Abstract Background Inflammatory bowel disease (IBD) is a chronic multifactorial gastrointestinal condition influenced by genetic, environmental, and immune system factors. Gut microbiota disruption plays a key role in disease onset and progression, leading to inflammation, impaired healing, and increased intestinal permeability1,2. Diet, particularly polyphenols from fruits, vegetables, and tea, can impact microbiota composition and offer anti-inflammatory benefits. This study explores the effects of these compounds on inflammation of the intestinal epithelium at the transcriptomic level. Methods A panel of polyphenolic compounds (n=8) was tested on Caco-2 cells, which were pre-treated with the metabolites before inflammation was induced for 24 h using TNF-α and IFN-γ. The effects were evaluated through targeted gene expression analysis using RT-qPCR. Prioritized compounds – Urolithin A (UroA) and Gallic Acid (GA) – were further tested on inflamed 3D colonic epithelial organoids derived from ulcerative colitis patient (UC) (n=1) and non-IBD individual (n=1), with mRNA sequencing performed on the NextSeq 550 (Illumina). Results Initial metabolite screening in Caco-2 cells was performed by analyzing the expression changes of pro-inflammatory cytokines-coding genes TNF-α, MCP-1, IL-8. Results demonstrated significant induction of cellular inflammation (p < 0.05) after exogenous stimullation with TNF-α and IFN-γ, which was most efficiently and consistenly reduced by GA and UroA. Therefore, GA and UroA were further selected for deeper evaluation on inflamed colonic epithelial organoids. RNA-seq revealed more pronounced cell reactivity to all treatments in UC-organoids compared to non-IBD-organoids and clear sample clustering based on treatment condition. DEA and GSEA of significantly dysregulated genes revealed interesting functional insights into potential mechanisms of action of UroA and GA. Importanly, in inflammed UC-organoids, pre-treatment with GA showed significant (FDR < 0.05) upregulation of pathways involved in cell junction assembly, and significant (FDR < 0.05) downregulation of pathways involved in metabolic processes. Together, these results suggest that GA may lead to colonic epithelial cell metabolic reprogramming in UC from energy-intensive and nucleotide-demanding processess and contribute to the resolution of inflammation and restoration of gut epithelial integrity. Conclusion In conclusion, during colonic inflammation, dietary polyphenol metabolites regulate inflammatory responses, epithelial barrier function, and energy production pathways at the transcriptomic level. Study was funded by the European Union and the State Secretariat for Education, Research and Innovation (SERI) (project: miGut-Health, grant no. 101095470). References 1 Jostins, L., et al. (2012). Host–microbe interactions have shaped the genetic architecture of inflammatory bowel disease. Nature, 491(7422), 119-124. https://doi.org/10.1038/nature11582 2 Cleynen, I., et al. (2016). Inherited determinants of Crohn’s disease and ulcerative colitis phenotypes: a genetic association study. The Lancet, 387(10014), 156-167. https://doi.org/10.1016/S0140-6736(15)00465-1

Aspergillus fumigatus is responsible for inflammation in a murine model of chronic obstructive pulmonary disease exacerbation

BackgroundIn patients with chronic obstructive pulmonary disease (COPD), a sensitization to A. fumigatus has been related to a decline in lung function, but the role of fungal agents in the disease pathogenesis remains unclear. The main purpose of the present study was to investigate whether cell inflammation could worsen after exposure to A. fumigatus spores in vitro and then, in mice, following chronic exposure to cigarette smoke mimicking COPD.MethodsThe inflammatory response to cigarette smoke alone or with A. fumigatus was investigated in cell culture models of murine macrophages and alveolar epithelial cells. In an animal model, mice were exposed daily to two cigarettes smoke over 14 weeks, and two intranasal instillations of 105 spores at weeks 7 and 14. Then, their lungs were recovered to perform inflammatory and histopathological analyses.ResultsIn co-cultures of macrophages and epithelial cells treated with both cigarette smoke extracts (CSE) and A. fumigatus compared to CSE alone there were significant inductions in TNF-α (6.2-fold) and CXCL-2 (21.5-fold) gene expression, confirmed by significant increases in the corresponding protein secretion. In the murine model, histological analyses of the lung after chronic smoke exposure showed an increase in airspace enlargement. Moreover, a Bio-Plex approach on bronchoalveolar lavage of cigarette smoke and A. fumigatus-treated mice showed significant increases in multiple inflammatory proteins secreted in the lung.ConclusionsThere was a stronger inflammatory response after cigarette smoke exposure with A. fumigatus compared to cigarette smoke alone. These findings were correlated with histopathological changes in the mouse lung in vivo.

Research on RTD Fluxgate Induction Signal Denoising Method Based on Particle Swarm Optimization Wavelet Neural Network.

Aeromagnetic surveying technology detects minute variations in Earth's magnetic field and is essential for geological studies, environmental monitoring, and resource exploration. Compared to conventional methods, residence time difference (RTD) fluxgate sensors deployed on unmanned aerial vehicles (UAVs) offer increased flexibility in complex terrains. However, measurement accuracy and reliability are adversely affected by environmental and sensor noise, including Barkhausen noise. Therefore, we proposed a novel denoising method that integrates Particle Swarm Optimization (PSO) with Wavelet Neural Networks, enhanced by a dynamic compression factor and an adaptive adjustment strategy. This approach leverages PSO to fine-tune the Wavelet Neural Network parameters in real time, significantly improving denoising performance and computational efficiency. Experimental results indicate that, compared to conventional wavelet transform methods, this approach reduces time difference fluctuation by 23.26%, enhances the signal-to-noise ratio (SNR) by 0.46%, and improves sensor precision and stability. This novel approach to processing RTD fluxgate sensor signals not only strengthens noise suppression and measurement accuracy but also holds significant potential for improving UAV-based geological surveying and environmental monitoring in challenging terrains.

Organocatalytic Activation of Alkynes Enabled Remote Control of Atroposelectivity via Vinylidene para-Quinone Methides.

Axially chiral o-VQMs have been extensively investigated as key intermediates to approach miscellaneous chiral structures. By sharp contrast, their structural isomers p-VQMs have not been previously documented. The major reason, which results in the significant delay, may ascribe to the inherent challenges in the enantioselective activation of alkynes in a remote manner. Herein, we demonstrate that the remote activation mechanism of para-hydroxyl-substituted arylacetylenes enables significant stereochemical induction, resulting in axially chiral aryl-alkenes with excellent enantiopurities. A series of control experiments are performed to elucidate the insights of this asymmetric transformation and to verify the involvement of multimolecular CPAs in the reaction process. These findings are expected to unlock a new feature for VQM chemistry and inspire investigation into the organocatalytic remote control of stereoselectivity.

Liver oval cells in response to HDAC1 inhibitor trichostatin A: immunohistochemical characterization using OV-6 hepatic expression.

Liver regeneration is intricate, involves many cells, and necessitates extended research. This study aimed to investigate the response of liver oval cells (bipotent liver progenitors) to the epigenetic modifier trichostatin A (TSA), an HDAC1 inhibitor, and to develop a scoring system for assessing the response of these cells. Three groups of equally divided rats (n=24) were selected: control (A, dimethyl sulfoxide treated); oval cell induction (B, acetylaminofluorene [2-AAF] to block hepatocyes/carbon tetrachloride [CCL4] to induce oval cell response); and epigenetic modulation (C, TSA post 2-AAF/CCL4 injury). The oval cell response was quantified using immunoreactivity to the OV-6 antibody, and the ductular response was measured by calculating the bile duct (BD) to portal vein (PV) ratio and the percentage of individual oval cells in liver sections. The expression level of HDAC1 was also analyzed. The administration of TSA significantly enhanced oval cell proliferation and the ductular response (6.13±0.28). The control group exhibited limited immunoreactivity to OV-6, while group B showed significant induction of ductular response with distinct morphology (4.13±0.28). The expression levels of HDAC1 were elevated in both the oval cell induction group and the epigenetic modulation group compared to the control group. This study developed a precise method for quantifying liver oval cells and analyzing their response to TSA. TSA administration enhanced oval cell proliferation, suggesting its significance in regulating hepatic progenitor cell dynamics. The findings support the use of epigenetic modifiers in liver regeneration and propose a scoring system for assessing the response of liver oval cells.

Canonical Wnt signalling from the area opaca induces and maintains the marginal zone in pre-primitive-streak stage chick embryos.

In chick embryos before primitive streak formation, the outermost extra-embryonic region, known as the area opaca (AO), was generally thought to act only by providing nutrients and mechanical support to the embryo. Immediately internal to the AO is a ring of epiblast called the marginal zone (MZ), separating the former from the inner area pellucida (AP) epiblast. The MZ does not contribute cells to any part of the embryo but is involved in determining the position of primitive streak formation from the adjacent AP epiblast. Recently, it was discovered that the AO can induce an MZ from AP epiblast. Here, we explore the nature of this inductive signal. We find that WNT8C is highly expressed in the AO, whereas canonical Wnt pathway targets are enriched in the MZ, along with strong nuclear β-catenin localization. Using isolation and recombination experiments combined with gain- and loss-of-function by exogenous chemical modulators of the pathway, we reveal that Wnt signalling is essential for induction and maintenance of the MZ, as well as sufficient to induce MZ properties in AP epiblast. We propose that canonical Wnt signalling is responsible for induction of the MZ by the area opaca.

Replisomal coupling between the α-Pol III core and the τ-subunit of the clamp loader complex (CLC) are essential for genomic integrity in E. coli.

Coupling interactions between the alpha (α) subunit of the polymerase III core (α-Pol III core) and the tau (τ) subunit of the clamp loader complex (τ-CLC) are vital for efficient and rapid DNA replication in Escherichia coli (E. coli). Specific and targeted mutations in the C-terminal τ-interaction region of the Pol III α-subunit disrupted efficient coupled rolling circle DNA synthesis in vitro and caused significant genomic defects in CRISPR-Cas9 dnaE edited cell strains. These α-Pol III mutations eliminated the interaction with τ-CLC but retained wild-type polymerase and exonuclease activities. The most severely affected mutant strains, dnaE:Y1119A and dnaE:L1097/8S, had significantly reduced doubling times, reduced fitness, and increased cellular length phenotypes as a result of this targeted decoupling of the replisome and the generation of replication stress. Those strains also showed significant SOS induction from unwound but unreplicated regions within the genome. In support, significant single-stranded DNA (ssDNA) gaps were detected by fluorescence microscopy and quantified by fluorescence activated cytometry using an in situ PLUG assay for those dnaE:mut strains. By comparing the biochemical and genomic consequences of disrupting the τ-CLC - α-Pol III coupling contacts, we have unveiled a more cohesive picture and mechanistic understanding of replisome dynamics and the essential interactions required to maintain overall fitness through a stable genome.

A fluidic device for continuous on-line inductive sensing of proteolytic cleavages.

Proteases, an important class of enzymes that cleave proteins and peptides, carry a wealth of potentially useful information. Devices to enable routine and cost effective measurement of their activity could find frequent use in clinical settings for medical diagnostics, as well as some industrial contexts such as detecting on-line biological contamination. In particular, devices that make use of readouts involving magnetic particles may offer distinct advantages for continuous sensing because material they release can be magnetically captured downstream and their readout is insensitive to optical properties of the sample. Bioassays based on giant magnetoresistance sensors that detect the binding or release of magnetic materials have been widely explored for these reasons, but they typically require expensive consumables. Here, we develop a simpler protease sensor based on inductive detection of particle release with pulsed magnetic fields, leveraging a design that incorporates both the pulse coil and gradiometer coils into a printed circuit board. Our fluidic chips are formed from casts of 3D printed molds, such that both the sensor and the consumable components could be relatively easy to mass produce. Using pulses ranging up to 10 s of mT, we show that our device has a limit of detection below 1 μg of iron and that its duty cycle can be varied to control temperature through Joule heating. By chemically functionalizing the glass surface of our fluidic chips with zwitterionic polymer and incorporating a PEG block co-polymer into the PDMS component, we are able to suppress the nonspecific binding of albumin by 7.8 times inside the chips. We demonstrate a layer-by-layer approach for covalently linking magnetic nanoparticles to the chips via cleavable peptide substrates. Finally, we observe the release of the magnetic particles from the chips under conditions of proteolytic cleavage and measure resulting changes in inductive signals, demonstrating a detection sensitivity for chymotrypsin in the hundreds of nM. The methods we establish here have the potential to aid progress toward sensors comprised of disposable fluidic chips measured by inexpensive detection devices that may one day facilitate ubiquitous protease activity monitoring.

Design and Detection Performance of Metal Contaminant Sensor with Triple Coil Structure

In marine engineering, the daily management of mechanical equipment needs to ensure that the oil is normal. Oil plays the role of sealing, cooling, lubrication, and other functions in the equipment, and can also be used as hydraulic fluid to transfer energy. By analyzing the state of the oil, it is possible to obtain information about the operation of the equipment, such as judging the wear or failure of the equipment by detecting impurities in the oil. This paper proposes and designs a wireless triple-coil structure oil detection sensor for detecting metal particles in the oil circuit. The sensor consists of three coils placed concentrically with the same parameters. When the sensor detects metal particles in the oil, the ferromagnetic and non-ferromagnetic particles flowing through the sensor produce magnetization and eddy current effects, resulting in variable inductive signals that complete the detection of metal particles. This paper firstly explains the sensing principle of this triple coil sensor detection by formula derivation. Secondly, the simulation model of the sensor was established by using COMSOL 6.0 simulation software according to the scale of 1:1, and the magnetic field strength distribution law inside the coil of the triple-coil sensor was simulated. The experimental results showed that the sensor was able to detect iron particles at 73 µm and copper particles at 220 µm, moreover the obtained signal characteristics are obvious, with high detection sensitivity. The sensor is wireless and performs contactless detection of metal particles. This is important for the detection of metal particle contaminants in oil.

Overexpression and purification of YidRv gene from the hypervirulent Klebsiella pneumoniae, and the ability of the gene product in inducing a humoral response

The yidRv gene, isolated from the hypervirulent Klebsiella pneumoniae (hvKP), is a novel gene with an unknown function; however, it has exhibited high homology to the yidR, a gene recognized as potential vaccine candidate. The aim of this study was to clone the yidRv gene from the Indonesian hvKP and to investigate its overexpression in Escherichia coli. In the experiment, yidRv was cloned into pET21 to construct pYik23. Recombinant protein YidRv was produced by growing E. coli BL21 (DE3)/pYik23 in LB medium with ampicillin at 29 °C, inducing protein synthesis with 0.5 mM IPTG for 20 hours. Purification was performed using Ni‐NTA resin, and the purified protein (50 µg) was administered to BALB/c mice to test for the production of IgG, IgM and IgA on 2 days before and day 19th and 37th after the first vaccination. The results show a significant induction of IgG and IgM, but not of IgA antibodies. In conclusion, the yidRv gene was overexpressed in E. coli BL21 (DE3) at high levels in soluble form, with the recombinant protein able to be purified to 90% homogeneity. The recombinant YidRv demonstrated the ability to stimulate the generation of both IgM and IgG antibodies.

Technical Code Analysis of Geomagnetic Flaw Detection of Suppression Rigging Defect Signal Based on Convolutional Neural Network

In this paper, technical code analysis and recognition of the defect signal of the suppression rigging based on a convolutional neural network are carried out given the difficulty and low recognition rate of the defect detection and recognition of the suppression rigging. Firstly, the magnetic induction signal of the suppression rigging defects is collected using CM-801 (Anshan, China), Kalman filtering is used to screen and pre-process the collected data, and the noise reduction data are presented in the form of a cloud image. The pressed rigging defect data set is constructed, and the region of broken wire defect and stress in the image is calibrated. The single-stage object detection algorithm YOLOv5 (You Only Look Once) based on convolutional neural network model calculation is used, the scale detection layer and positioning loss function of the YOLOv5 algorithm are improved and optimized, and the improved YOLOv5 algorithm is used for experiments. The experimental results show that the detection accuracy of the convolution neural network model can reach 97.1%, which can effectively identify the defect signal of the suppressed rigging.

Species-specific differences in acetaminophen hepatotoxicity depend on HSP70 expression level.

Acetaminophen (N-Acetyl-p-aminophenol: APAP) is one of the most commonly used analgesic/antipyretic drugs with proven safety at therapeutic doses, however, over-dosage causes dose-dependent liver damage, leading to acute liver failure in severe cases. The level of APAP-induced liver injury has been known to vary among animal species, and APAP concentrations that induce cell death have been investigated using primary cultured cells. We constructed in vitro model of APAP-induced hepatotoxicity using mouse, rat, and human hepatoma cell lines to investigate species differences in the APAP-induced cytotoxicity by monitoring cell death as a marker. The EC50 for each cell line was Hepa1-6 (mouse) < H-4-II-E (rat) < Hep3B (human), while the expression of heat shock protein 70 (HSP70), which was a typical molecular chaperone, positively correlated with the EC50 of each cell. Heat shock treatment, which caused activation of heat shock factor 1 (HSF1) followed by significant induction of HSP70, partially suppressed APAP-induced cell death in Hepa1-6 and H-4-II-E. Moreover, HSP70 or HSF1 siRNA treatment in Hep3B enhanced APAP-induced cell death. These results suggest that APAP-induced cell death in hepatoma cell lines may be partly mediated by protein denaturation and that the expression level of HSP70 has an inhibitory effect.

Research on the Detection Method for Feeding Metallic Foreign Objects in Coal Mine Crushers Based on Reflective Pulsed Eddy Current Testing

In response to the difficulties and poor timeliness in detecting feeding metallic foreign objects during high-yield continuous crushing operations in coal mines, this paper proposes a new method for detecting metallic foreign objects, combining pulsed eddy current testing with the Truncated Region Eigenfunction Expansion (TREE) method. This method is suitable for the harsh working conditions in coal mine crushing stations, which include high dust, strong vibration, strong electromagnetic interference, and low temperatures in winter. A model of the eddy current field of feeding metallic foreign objects in the truncated region is established using a coaxial excitation and receiving coil with a Hall sensor. The full-cycle time-domain analytical solution for the induced voltage and magnetic induction intensity of the reflective field under practical square wave signals is obtained. Simulation and experimental results show that the effective time range, peak value, and time to peak of the received voltage and magnetic induction signals can be used to classify and identify the size, thickness, conductivity, and magnetic permeability of feeding metallic foreign objects. Experimental results meet the actual needs for removing feeding metallic foreign objects in coal mine sites. This provides core technical support for the establishment of a predictive fault diagnosis system for crushing equipment.

Kupffer cells, the limelight in the liver regeneration.

Kupffer cells are pivotal in initiating hepatocyte proliferation and establishing connections between different cell types during liver regeneration following partial hepatectomy. As resident macrophages within the liver, Kupffer cells collaborate with hepatocytes and non-parenchymal cells to release various inflammatory mediators that promote hepatocyte proliferation through induction signals like STAT3 phosphorylation. Additionally, the regeneration and replenishment of Kupffer cells themselves are integral components of liver regeneration. The supplementation of the Kupffer cell pool primarily occurs through two pathways: one involves local proliferation of Kupffer cells in their original location, while the other entails infiltration of circulating monocytes into the liver, followed by acquiring Kupffer cell phenotypes under the combined influence of multiple inducing factors. Extensive research has focused on intercellular crosstalk among various types of liver cells during liver regeneration, highlighting the crucial role played by Kupffer cells. This article aims to introduce Kupffer cells and their involvement in liver regeneration, as well as discuss the steady-state balance of Kupffer cell pools during this process.

Combination of chemotherapy and c-MET inhibitors has synergistic effects in c-MET overexpressing pancreatic cancer cells.

Pancreatic ductal adenocarcinoma (PDAC) remains as one of the most lethal malignancies. c-MET is an important oncogenic kinase involved in PDAC progression. We determined the anticancer effect of c-MET inhibitors, crizotinib and cabozantinib, combined with chemotherapeutic agents, doxorubicin, oxaliplatin and gemcitabine, against different PDAC and a lung adenocarcinoma cell line expressing different levels of c-MET. MTT assay was performed to assess cell growth inhibition. Synergistic combinations were evaluated in spheroid cultures, while apoptosis was determined through Hoechst33258 staining. The effect of drug combinations on cell cycle and apoptosis induction was examined by RNase/PI flow cytometric assay. We also evaluated reactive oxygen species (ROS) levels using 2',7'-dichlorofluorescein-diacetate (DCFH-DA) assay to explore the possible mechanisms contributing to synergism. Combination of crizotinib or cabozantinib with doxorubicin exhibited synergistic effects in c-MET overexpressing cells. Conversely, combinations of c-MET inhibitors with other agents were additive or even antagonistic. Combination index (CI) values calculated with Calcusyn software were 0.631-0.730 for crizotinib and 0.542-0.746 for cabozantinib co-administrations. Synergistic combinations showed significant spheroid growth inhibition and apoptosis induction in Suit-2, c-MET dependent PDAC cells. These combinations also significantly increased the number of cells in both apoptotic sub-G1 phase and the G2/M phase compared to single-drug treatment. Increased ROS production seemed to be a possible mechanism underlying synergism. In conclusion, c-MET inhibitors synergize with DNA damaging agent, doxorubicin, in cancer cells with c-MET overexpression, indicating that these combination therapies may be a promising cancer therapeutic strategy.

A Force-Thermal-Magnetic Trimodal Flexible Sensor for Ultrafine Recognition of Metallic Materials.

Humans possess the remarkable ability to perceive the intricate world by integrating multiple senses. However, the challenge of enabling humanoid robots to achieve multimodal sensing and fine recognition of metallic materials persists. In this study, we propose a flexible tactile sensor that mimics the sensory patterns of human skin, which is assembled by a flexible electromagnetic coil that is engraved on the surface of a polyimide substrate and porous MXene/CNT aerogel. This sensor is capable of detecting pressure, temperature, and inductive signals with minimal interference via three disparate response mechanisms of the piezoresistive sensing, the thermoelectric principle, and the electromagnetic induction effect, allowing the device with the abilities of sensing grasp forces and selectively identifying ferromagnetic and nonferromagnetic metals, which has a high accuracy rate of 99.2% in distinguishing mixed metals with varying ratios based on the fusion algorithm of multimodal sensory data. Further, the sensor was integrated on a humanoid robotic hand to demonstrate its recognition capacity of objects used in a kitchen setting and a simulated scenario of mineral exploration, achieving a remarkable ultrafine accuracy of 100% in distinguishing 16 common metal products. These findings will pave the way for humanoid robots to attain heightened levels of perception and recognition.

Fibroblast Growth Factor 8 enhances the chondrogenesis of trunk neural crest cells: a possible gene regulatory network.

The neural crest (NC) is an embryonic cell population with high migratory capacity. It contributes to forming several organs and tissues, such as the craniofacial skeleton and the peripheral nervous system of vertebrates. Both pre-migratory and post-migratory NC cells are plastic, adopting multiple differentiation paths by responding to different inductive environmental signals. Cephalic neural crest cells (CNCCs) give rise to most of the cartilage and bone tissues in the head. On the other hand, the mesenchymal potential of trunk neural crest cells (TNCCs) is sparsely detected in some animal groups. The mesenchymal potential of TNCCs can be unveiled through specific environmental conditions of NC cultures. In this study, we present evidence that FGF8 treatment can foster increased chondrogenic differentiation of TNCCs, particularly during treatment at the migratory stage. Additionally, we conducted a transcriptomic analysis of TNCCs in the post-migratory stage, noting that exogenous FGF8 signaling can sustain multipotent status and, possibly, at the same time, a pro-cartilage regulatory gene network. Our results provide a more comprehensive understanding of the mechanisms underlying chondrogenic differentiation from TNCCs.

Sperm-Borne Mitochondrial Activity Influenced by Season and Age of Holstein Bulls.

Sperm mitochondria are vital organelles for energy production and pre- and post-fertilization sperm functions. The potential influence of the age of the bull and season on the sperm-borne mitochondrial copy number and the transcription activity has not yet been investigated. Therefore, the expression patterns of all protein-coding mitochondrial genes were identified throughout the year along with mitochondrial copy numbers in young and old bulls' spermatozoa. For that, high-quality semen samples (n = 32) with more than 80% quality for the morphological parameters, from young (n = 4, aged 18-24 months old) and old (n = 4, aged 40-54 months old) Holstein bulls, were collected during the four seasons (n = 4 samples each animal/season). The DNA and RNA were isolated from sperm cells and subjected to the DNA copy number and expression analyses using qPCR. Furthermore, an in silico analysis using gene ontology online tools for the abundantly expressed genes was utilized. The data were statistically analyzed using Prism10 software. There was a significant reduction in the mitochondria copy number of young bulls' spermatozoa compared to their old counterparts during the summer (29 ± 3 vs. 51 ± 6, p < 0.001) and winter (27 ± 3 vs. 43 ± 7, p < 0.01) seasons. However, sperm-borne mitochondrial protein-coding genes were transcriptionally higher in young bulls throughout the year. Within the same group of bulls, unlike the old bulls, there was a significant (p < 0.05) induction in the transcription activity accompanied by a significant (p < 0.05) reduction in the mitochondrial copy numbers in the summer (29 ± 3) and winter (27 ± 3) compared to the spring (42 ± 9) and autumn (36 ± 5) seasons in young bulls. Additionally, the pathway enrichment of the top six expressed genes differed between age groups and seasons. In conclusion, under the same quality of semen, the early stages of age are associated with mitochondrial biogenesis and transcription activity dysregulation in a season-dependent manner.

The Sdp-SH3b2 domain contained in Lactobacillus johnsonii N6.2-derived extracellular vesicles inhibit murine norovirus replication.

The internalization of Lactobacillus johnsonii N6.2 extracellular vesicles (EVs) by cells results in a significant induction of the 2',5'-oligoadenylate synthetase (OAS) pathway. It also induces expression of IFI44L, MX1, MX2 and DDX60. In this work, we evaluated whether the antiviral response induced by L. johnsonii N6.2-derived EVs, has an inhibitory effect on an RNA viral insult using murine norovirus (MNV-1) as the viral infection model. We found that RAW 264.7 Macrophages treated with EVs significantly decreased the levels of MNV-1 genome. These results were consistent with an increase in expression of Oas1b, Oas2, Oasl, Mx1, Mx2 and Ifi44l (6 hours post infection). Out of six proteins enriched in EVs, we found that SH3b2 domain of Sdp was the only protein effector molecule able to recapitulate the activation of the OAS pathway. In C57BL6 mice, the administration of live L. johnsonii N6.2, EVs, and Sdp-SH3b2/liposomes significantly decreased MNV-1 titers in the distal ileum, in contrast to the controls with PBS and liposomes alone that did not affect MNV-1. These results establish that the SH3b2 domain of Sdp, which is enriched in L. johnsonii derived EVs, is an effector molecule in EVs that can orchestrate the control of viral infections in vivo.

Poly(allylamine)/tripolyphosphate nanocomplex coacervate as a NLRP3-dependent systemic adjuvant for vaccine development.

Nanotechnology plays a crucial role in vaccine development. It allows the design of functional nanoparticles (NPs) that can act both as antigen carriers and as adjuvants to enhance the immune response. The present study aims to evaluate complex coacervate-like NPs composed of poly(allylamine hydrochloride) (PAH) and tripolyphosphate (TPP) as a safe vehicle and adjuvant for systemic vaccines. We investigated the activation of different antigen-presenting cells (APCs) with NPs and their adjuvanticity in Balbc/c and different KO mice that were intraperitoneally immunized with NP-OVA. We found that NPs increased the expression of CD86 and MHCII and promoted the production and secretion of interleukin-1β (IL-1β) and IL-18 through the inflammasome NLRP3 when macrophages and dendritic cells were co-incubated with LPS and NPs. We evidenced an unconventional IL-1β release through the autophagosome pathway. The inhibition of autophagy with 3-methyladenine reduced the LPS/NPs-induced IL-1β secretion. Additionally, our findings showed that the systemic administration of mice with NP-OVA triggered a significant induction of serum OVA-specific IgG and IgG2a, an increased secretion of IFN-γ by spleen cells, and high frequencies of LT CD4 + IFN-γ + and LT CD8 + IFN-γ + . Our findings show that NPs promoted the inflammasome activation of innate cells with Th1-dependent adjuvant properties, making them valuable for formulating novel preventive or therapeutic vaccines for infectious and non-infectious diseases.