Time Course Research Articles

Variation in plant cover and flower production of Carex rostrata and C. vesicaria within a freshwater littoral zone. A 14-year time course

Variation in plant cover and flower production of Carex rostrata and C. vesicaria within a freshwater littoral zone. A 14-year time course

Disparity in temporal and spatial relationships between resting-state electrophysiological and fMRI signals.

Resting-state brain networks (RSNs) have been widely applied in health and disease, but the interpretation of RSNs in terms of the underlying neural activity is unclear. To address this fundamental question, we conducted simultaneous recordings of whole-brain resting-state functional magnetic resonance imaging (rsfMRI) and electrophysiology signals in two separate brain regions of rats. Our data reveal that for both recording sites, spatial maps derived from band-specific local field potential (LFP) power can account for up to 90% of the spatial variability in RSNs derived from rsfMRI signals. Surprisingly, the time series of LFP band power can only explain to a maximum of 35% of the temporal variance of the local rsfMRI time course from the same site. In addition, regressing out time series of LFP power from rsfMRI signals has minimal impact on the spatial patterns of rsfMRI-based RSNs. This disparity in the spatial and temporal relationships between resting-state electrophysiology and rsfMRI signals suggests that electrophysiological activity alone does not fully explain the effects observed in the rsfMRI signal, implying the existence of an rsfMRI component contributed by "electrophysiology-invisible" signals. These findings offer a novel perspective on our understanding of RSN interpretation.

Emotional attention: Time course and effects of agonistic and antagonistic overlay of intrinsic and goal relevances.

Emotional attention can be explained within a goal-directed theory framework according to which attention is captured by the goal relevance of stimuli, that is, their conduciveness nature to a momentarily important goal. However, such an explanation does not consider the attentional impact of intrinsic relevance of stimuli, that is, their general pleasantness. This problem could be resolved by appraisal theories, suggesting that attention is captured by intrinsic relevance and goal relevance of stimuli, whether the relevance overlay is agonistic (e.g., pleasant and goal-conducive) or antagonistic (e.g., unpleasant and goal-conducive). Moreover, appraisal theories suggest that early and late attentional capture would be more impacted by intrinsic relevance and goal relevance, respectively. In the present study, we confronted the predictions of appraisal theories with that of goal-directed theory. To this end, 120 participants performed parallelly an induction task to induce different relevance values to three colored squares, and a dot-probe task with two different stimulus onset asynchrony, to measure early and late attentional captures. This paradigm allowed us to measure attentional capture between a neutral stimulus, a goal-relevant stimulus, and an overlay stimulus. The overlay stimulus was agonistic in one group, while it was antagonistic in the other group. Our results showed evidence in favor of appraisal theories. Namely, the overlay stimulus captured more attention than the goal-relevant and the neutral stimulus, regardless of whether the overlay was agonistic or antagonistic. However, our results were mixed regarding the effects of intrinsic relevance and goal relevance on attentional capture as a function of temporality. (PsycInfo Database Record (c) 2024 APA, all rights reserved).

An Explainable and Generalizable Recurrent Neural Network Approach for Differentiating Human Brain States on EEG Dataset.

Electroencephalogram (EEG) is one of the most widely used brain computer interface (BCI) approaches. Despite the success of existing EEG approaches in brain state recognition studies, it is still challenging to differentiate brain states via explainable and generalizable deep learning approaches. In other words, how to explore meaningful and distinguishing features and how to overcome the huge variability and overfitting problem still need to be further studied. To alleviate these challenges, in this work, a multiple random fragment search-based multilayer recurrent neural network (MRFS-MRNN) is proposed to improve the differentiating performance and explore meaningful patterns. Specifically, an explainable MRNN module is proposed to capture the temporal dependences preserved in EEG time series. Besides, a MRFS module is designed to cut multiple random fragments from the entire EEG signal time course to improve the effectiveness of brain state differentiating ability. MRFS-MRNN is concatenatedto effectively overcome the huge variabilities and overfitting problems. Experiment results demonstrate that the proposed MRFS-MRNN model not only has excellent differentiating performance, but also has good explanation and generalization ability. The classification accuracies reach as high as 95.18% for binary classification and 89.19% for four-category classification on the individual level. Similarly, 95.53% and 85.84% classification accuracies are obtained for the binary and four-category classification on the group level. What's more, 94.28% and 85.43% classification accuracies of binary and four-category classifications are achieved for predicting brand new subjects. The experiment results showed that the proposed method outperformed other state-of-the-art (SOTA) models on the same underlying data and improved the explanation and generalization ability.

Reversible assembly and disassembly of V-ATPase during the lysosome regeneration cycle.

Regulation of the luminal pH of late endocytic compartments in continuously fed mammalian cells is poorly understood. Using normal rat kidney fibroblasts, we investigated the reversible assembly/disassembly of the proton pumping V-ATPase when endolysosomes are formed by kissing and fusion of late endosomes with lysosomes and during the subsequent reformation of lysosomes. We took advantage of previous work showing that sucrosomes formed by the uptake of sucrose are swollen endolysosomes from which lysosomes are reformed after uptake of invertase. Using confocal microscopy and subcellular fractionation of NRK cells stably expressing fluorescently tagged proteins, we found net recruitment of the V1 subcomplex during sucrosome formation and loss during lysosome reformation, with a similar time course to RAB7a loss. Addition of invertase did not alter mTORC1 signalling, suggesting that the regulation of reversible V-ATPase assembly/disassembly in continuously fed cells differs from that in cells subject to amino acid depletion/refeeding. Using live cell microscopy, we demonstrated recruitment of a fluorescently tagged V1 subunit during endolysosome formation and a dynamic equilibrium and rapid exchange between the cytosolic and membrane bound pools of this subunit. We conclude that reversible V-ATPase assembly/disassembly plays a key role in regulating endolysosomal/lysosomal pH in continuously fed cells.

Application of a U-Net Neural Network to the Puccinia sorghi-Maize Pathosystem.

Computer vision approaches to analyze plant disease data can be both faster and more reliable than traditional, manual methods. However, the requirement of manually annotating training data for the majority of machine learning applications can present a challenge for pipeline development. Here, we describe a machine learning approach to quantify Puccinia sorghi incidence on maize leaves utilizing U-Net convolutional neural network models. We analyzed several U-Net models with increasing amounts of training image data, either randomly chosen from a large data pool or randomly chosen from a subset of disease time course data. As the training dataset size increases, the models perform better, but the rate of performance decreases. Additionally, the use of a diverse training dataset can improve model performance and reduce the amount of annotated training data required for satisfactory performance. Models with as few as 48 whole-leaf training images are able to replicate the ground truth results within our testing dataset. The final model utilizing our entire training dataset performs similarly to our ground truth data, with an intersection over union value of 0.5002 and an F1 score of 0.6669. This work illustrates the capacity of U-Nets to accurately answer real-world plant pathology questions related to quantification and estimation of plant disease symptoms. [Formula: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Accessibility of working memory representations in the focus of attention: Heightened or reduced?

Many researchers agree that information residing in the focus of attention in working memory benefits from a boost in memory strength and activation, as well as heightened accessibility. However, recent studies have questioned this heightened accessibility. More specifically, these recent studies found reduced accessibility for an item in the focus of attention compared to another item in working memory, which was referred to as an "inhibition-of-return-like" effect. Our study aimed to provide a detailed examination of the accessibility of information in the focus of attention. Across a series of experiments, varying task characteristics related to the time course of the effect (Experiments 2-3) and the potential role of response inhibition (Experiments 4a-4b), we repeatedly failed to find evidence for an inhibition-of-return-like effect. Instead, we mostly found heightened accessibility for an item in the focus of attention. Given that an inhibition-of-return-like effect seems to be limited to a very specific task condition, reduced accessibility of information in the focus of attention appears to be far from a general phenomenon. Therefore, based on our findings, we propose that information in the focus of attention generally benefits from heightened accessibility, although there may be factors, most likely unrelated to the functioning of the focus of attention, that could sometimes mask this and even result in data patterns that are consistent with reduced accessibility. The theoretical implications for the focus of attention and working memory are discussed. (PsycInfo Database Record (c) 2024 APA, all rights reserved).

تبلیغی جماعت کے ذرائع تبلیغ کا تحقیقی جائزہ

Preaching of Islam is the most important responsibility of Ummah. For the payment of this duty various missionary organizations were established in the Muslim world. Tablighi Jammat is also a Tablighi movement, which was established by Maulana Muhammad Ilyas. Because the Maio nation was in danger of apostasy, this requires a Tablighi Jammat occurred. Now this movement is active in many countries of the world. Tablighi Jammat has own ways and timing course for preaching. They use different ways to preach. They preach through speeches, letters. They also send Tablighi deputations to entire country and outside countries for preaching. Besides male parties’ female parties also send. On the other hand, they use modern resources for preaching such as TV, Internet, Laptop, videos and audios etc. These means easy and useful to conveying the message of Islam.

Dampak Perkembangan Teknologi Terhadap Mata Pencaharian Masyarakat

In our lives at this time of course we will not be separated by technology. Because it is an effort for humans to meet the needs of their lives both in terms of quality and quantity. Technology has many impacts on human life, both negative and positive impacts. This research was conducted in Dolopo Village, Dolopo District, Madiun regency, East Java province. The formulation of the problems that will be discussed in this study include: 1) the response of the Dolopo community to the entry of technology into people’s livelihoods. 2) on the impact of the use of technology on the socio-economic community in Dolopo Village. The type of research used is field research, while data collection techniques are carried out by observation, and interviews. The results of this study concluded that the impact of technological developments facilitate human activities but on the other hand also lead to loss of community livelihoods.

Numerical Simulation on Dispersion of Hydrogen Leaked in Particle Layers of Glass Beads and Soil

In order to understand the dispersion of hydrogen (H2) leaked in packed beds of nonporous/porous particles in a partially open space practically, the dispersion of H2 in the particle layers of glass beads and soil was analytically studied using a computational fluid dynamics code to be compared with the experiments and to elucidate the effects of the particle layer. The packed beds in the partially open space can be considered as a basic model for all processes of transfer, treatment, storage, and disposal of radioactive materials containing fuel debris in the decommissioning of nuclear facilities after a severe accident. The H2 flowing out from a single leak point in the particle layer of nonporous glass beads was affected by buoyancy around the leak point and diffused directly above the leak point in an elliptical shape faster than in the horizontal direction. After that, when it reached the air layer in the head space above the particle layer, the H2 spread horizontally, formed a large concentration gradient near the boundary between the particle layer and the air layer, and further diffused in the air layer until the H2 concentration became about 1/3 or less of the concentration near the surface of the particle layer. When the particle diameters were 512 μm and 1200 μm, this tendency was more pronounced when the porosity was the same and the particle diameter was smaller. The calculations largely reproduced the experimental concentration distributions. When the particle layer was porous decomposed granite soil, the diffusion behavior of H2 in the particle layer proceeded in the same manner as in the case of the glass beads. However, a large concentration gradient was formed near the boundary between the particle layer and the air layer, and then H2 diffused in the air layer until the H2 concentration became below the lower combustion limit. It was suggested through sensitivity analysis that the air permeability coefficient had a large effect on the time course of the H2 concentration distribution. Based on this information, we further simulated H2 behavior in the vessel containing the H2 leaked particle layer. By inserting multiple vent pipes without considering H2 generation distribution and particle properties in the particle layer, H2 accumulated from one pipe was discharged by buoyancy without depending on the H2 generation distribution and particle properties in the particle layer and air flowed in from the other pipe. It was suggested that such a natural ventilation process would reduce the H2 concentration in the container.

Power output at the moderate-to-heavy intensity transition decreases in a non-linear fashion during prolonged exercise.

The aims of this study were to: (i) describe the time course of the decrease in power output at the moderate-to-heavy intensity transition during prolonged exercise; (ii) investigate the association between durability of the moderate-to-heavy intensity transition and exercise capacity; and (iii) explore physiological correlates of durability of the moderate-to-heavy intensity transition. Twelve trained cyclists (age: 40 ± 8y, O2peak: 52.3 ± 5.2mL·min-1·kg-1) performed an exhaustive cycling protocol involving alternating incremental exercise tests to determine power output at the moderate-to-heavy intensity transition via the first ventilatory threshold (VT1), and 30-min bouts at 90% of the power output at the previously estimated VT1 in the rested state. The individual time course of VT1 was modelled using linear and second-order polynomial functions, and time to a 5% decrease in VT1 (Δ5%VT1) was estimated using the best-fitting model. Power output at VT1 decreased according to a second-order polynomial function in 11 of 12 participants. Time-to-task failure (234 ± 66 min) was correlated with Δ5%VT1 (139 ± 78 min, rs = 0.676, p = 0.016), and these were strongly correlated with absolute and relative rates of fat oxidation at specific exercise intensities measured during the incremental test performed in the rested state. These data: (i) identify a non-linear time course of decreases in the moderate-to-heavy intensity transition during prolonged exercise; (ii) support the importance of durability of the moderate-to-heavy intensity transition in prolonged exercise capacity; and (iii) suggest durability of the moderate-to-heavy intensity transition is related to fat oxidation rates.

Large-scale bioreactor production of extracellular vesicles from mesenchymal stromal cells for treatment of acute radiation syndrome

BackgroundHematopoietic acute radiation syndrome (H-ARS) occurring after exposure to ionizing radiation damages bone marrow causing cytopenias, increasing susceptibility to infections and death. We and others have shown that cellular therapies like human mesenchymal stromal cells (MSCs), or monocytes/macrophages educated ex-vivo with extracellular vesicles (EVs) from MSCs were effective in a lethal H-ARS mouse model. However, given the complexity of generating cellular therapies and the potential risks of using allogeneic products, development of an “off-the-shelf” cell-free alternative like EVs may have utility in conditions like H-ARS that require rapid deployment of available therapeutics. The purpose of this study was to determine the feasibility of producing MSC-derived EVs at large scale using a bioreactor and assess critical quality control attributes like identity, sterility, and potency in educating monocytes and promoting survival in a lethal H-ARS mouse model.MethodsEVs were isolated by ultracentrifugation from unprimed and lipopolysaccharide (LPS)-primed MSCs grown at large scale using a hollow fiber bioreactor and compared to a small scale system using flasks. The physical identity of EVs included a time course assessment of particle diameter, yield, protein content and surface marker profile by flow-cytometry. Comparison of the RNA cargo in EVs was determined by RNA-seq. Capacity of EVs to generate exosome educated monocytes (EEMos) was determined by qPCR and flow cytometry, and potency was assessed in vivo using a lethal ARS model with NSG mice.ResultsPhysical identity of EVs at both scales were similar but yields by volume were up to 38-fold more using a large-scale bioreactor system. RNA-seq indicated that flask EVs showed upregulated let-7 family and miR-143 micro-RNAs. EEMos educated with LPS-EVs at each scale were similar, showing increased gene expression of IL-6, IDO, FGF-2, IL-7, IL-10, and IL-15 and immunophenotyping consistent with a PD-L1 high, CD16 low, and CD86 low cell surface expression. Treatment with LPS-EVs manufactured at both scales were effective in the ARS model, improving survival and clinical scores through improved hematopoietic recovery. EVs from unprimed MSCs were less effective than LPS-EVs, with flask EVs providing some improved survival while bioreactor EVs provide no survival benefit.ConclusionsLPS-EVs as an effective treatment for H-ARS can be produced using a scale-up development manufacturing process, representing an attractive off-the-shelf, cell-free therapy.

Bending creep behaviour of various polymer films analysed by surface strain measurement.

Understanding the temporal bending deformation of polymer films is key to designing mechanically durable flexible electronic devices. However, such creep behaviour under persistent bending remains unclear due to a lack of precise and accurate bending strain analysis methods. Herein, we quantitatively analysed the bending creep behaviour of various polymeric films using our developed strain measurement method that can precisely measure surface strain from optical diffraction. The surface strain measurement reveals that bending creep deformation differs depending on the polymer structure. The four-element Burgers model was employed to model the temporal strain increase on the bending surface successfully. By fitting the four-element model to the time course of the measured surface strain, we found that each polymer film has a different threshold surface strain for the appearance of bending creep deformation. Such disparity in the bending creep behaviour can be explained by the difference in strain energy density between the polymer films and their elastic model; polymer films with a small strain energy density difference show small bending creep deformation. The results obtained in this study contribute to the elucidation of the bending creep behaviour of polymer films and the development of flexible electronic devices operated under persistent bending.

Maturation of cortical input to dorsal raphe nucleus increases behavioral persistence in mice.

The ability to persist towards a desired objective is a fundamental aspect of behavioral control whose impairment is implicated in several behavioral disorders. One of the prominent features of behavioral persistence is that its maturation occurs relatively late in development. This is presumed to echo the developmental time course of a corresponding circuit within late-maturing parts of the brain, such as the prefrontal cortex, but the specific identity of the responsible circuits is unknown. Here, we used a genetic approach to describe the maturation of the projection from layer 5 neurons of the neocortex to the dorsal raphe nucleus in mice. Using optogenetic assisted circuit mapping, we show that this projection undergoes a dramatic increase in synaptic potency between postnatal weeks 3 and 8, corresponding to the transition from juvenile to adult. We then show that this period corresponds to an increase in the behavioral persistence that mice exhibit in a foraging task. Finally, we used a genetic targeting strategy that primarily affected neurons in the medial prefrontal cortex (mPFC), to selectively ablate this pathway in adulthood and show that mice revert to a behavioral phenotype similar to juveniles. These results suggest that frontal cortical to dorsal raphe input is a critical anatomical and functional substrate of the development and manifestation of behavioral persistence.

Management of inorganic elements by overwintering physiology of cold hardy larvae of European corn borer (Ostrinia nubilalis, Hbn.).

The European corn borer (Ostrinia nubilalis, Hbn.), enters diapause, a strategy characterized by arrest of development and reproduction, reduction of metabolic rate and the emergence of increased resistance to challenging seasonal conditions as low sub-zero winter temperatures. The aim of this study was to investigate the potential role of inorganic elements in the ecophysiology of O. nubilalis, analysing their content in the whole body, hemolymph and fat body, both metabolically active, non-diapausing and overwintering diapausing larvae by ICP-OES spectrometer following the US EPA method 200.7:2001. O nubilalis as many phytophagous lepidopteran species maintain a very low extracellular sodium concentration and has potassium as dominant cation in hemolymph of their larvae. Changes in hemolymph and the whole body sodium content occur already at the onset of diapause (when the mean environmental temperatures are still high above 0ºC) and remain stable during the time course of diapause when larvae of this species cope with sub-zero temperatures, it seems that sodium content regulation is rather a part of diapausing program than the direct effect of exposure to low temperatures. Compared to non-diapausing O. nubilalis larvae, potassium levels are much higher in the whole body and fat body of diapausing larvae and substantially increase approaching the end of diapause. The concentration of Ca, Mg, P and S differed in the whole body, hemolymph and fat body between non-diapausing and diapausing larvae without a unique trend during diapause, except an increase in their contents at the end of diapause.

Using Irradiation-Acquisition Interleaved Time-Integrated Imaging to Assess Delayed Luminescence and the Noise-Laden Residual Spontaneous Photon Emission of Yeast

Delayed luminescence from organisms informs oxidative stress that may be modulable by external stimulations. In the absence of external stress causing delayed luminescence, organisms may produce spontaneous ultraweak photon emission due to the residual oxygen demand. To better understand the oxidative state of an organism, it is desirable to acquire the delayed luminescence to reach the phase wherein the ultraweak photon emission resides. This, however, is challenging due to the significant difference in the order of magnitude of the photon counts between the two types of photon emission. Conventional time-gated measurement requires a high dynamic range to assess the noise-level photon emission, whereas simple long exposure can miss the kinetics of luminescence. There may be a compromise to be made between robustly acquiring the decay kinetics of the delayed luminescence and reliably acquiring the noise-laden spontaneous photon emission. We demonstrate an irradiation-acquisition interleaved time-integrated imaging approach that may enable the reliable acquisition of slow-decay delayed luminescence down to the level of ultraweak photon emission. Repetitive irradiation was interleaved with a gradually increased time of acquisition to assess the integrated time course of the post-irradiation luminescence. Such instrument configuration performed on yeast facilitated the use of time differentiation to assess the delayed luminescence down to the noise-level ultraweak photon emission at the expense of the total time of acquisition.

Sarcomeric tropomyosin expression during human iPSC differentiation into cardiomyocytes.

Tropomyosin (TPM) is an essential sarcomeric component, stabilizing the thin filament and facilitating actin's interaction with myosin. In mammals, including humans, there are four TPM genes (TPM1, TPM2, TPM3, and TPM4) each of which generates a multitude of TPM isoforms via alternative splicing and using different promoters. In this study, we have examined the expression of transcripts as well as proteins of various sarcomeric TPM isoforms during human inducible pluripotent stem cell differentiation into cardiomyocytes. During the differentiation time course, we harvested cells on Days 0, 5, 10, 15, and 20 to analyze for various sarcomeric TPM transcripts by qRT-PCR and for sarcomeric TPM proteins using two-dimensional Western blot with sarcomeric TPM-specific CH1 monoclonal antibody followed by mass spectra analyses. Our results show increasing levels of total TPM transcripts and proteins during the period of differentiation, but varying levels of specific TPM isoforms during the same period. By Day 20, the rank order of TPM transcripts was TPM1α > TPM1κ > TPM2α > TPM1μ > TPM3α > TPM4α. TPM1α was the dominant protein produced with some TPM2 and much less TPM1κ and μ. Interestingly, small amounts of two lower molecular weight TPM3 isoforms were detected on Day 15. To the best of our knowledge this is the first demonstration of TPM1μ non-muscle isoform protein expression before and during cardiac differentiation.

Model-based simulation of glycaemic effect and body weight loss when switching from semaglutide or dulaglutide to once weekly tirzepatide

Objective To evaluate the efficacy endpoints of HbA1c and body weight loss after switching from the GLP-1 receptor agonists, semaglutide or dulaglutide, to treatment with the GIP/GLP-1 receptor agonist (RA) tirzepatide. Methods Models were developed and validated to describe the HbA1c and weight loss time course for semaglutide (SUSTAIN 1-10), dulaglutide (AWARD-11) and tirzepatide (SURPASS 1-5, phase 3 global T2D program). The impact of switching from once weekly GLP-1 RAs to tirzepatide was described by simulating the efficacy time course. Semaglutide and dulaglutide doses were escalated in accordance with their respective labels. Results Model-predicted mean decreases from baseline in HbA1c and body weight for semaglutide 0.5 mg, 1 mg, and 2 mg were 1.22 to 1.79% and 3.62 to 6.87 kg respectively, at Week 26. Model-predicted mean decreases from baseline in HbA1c and body weight for dulaglutide 1.5 mg, 3 mg and 4.5 mg were 1.53 to 1.84% and 2.55 to 3.71 kg respectively, at Week 26. After switching to tirzepatide 5, 10 and 15 mg HbA1c reductions were predicted to range between 1.95 to 2.46% and body weight reductions between 6.50 to 12.1 kg by Week 66. Conclusion In this model-based simulation, switching from approved maintenance doses of semaglutide or dulaglutide to tirzepatide, even at the lowest approved maintenance dose of 5 mg, showed the potential to further improve HbA1c and body weight reductions.

Persistence of Contact Lens-Induced Corneal Parainflammation Following Lens Removal.

Contact lens wear induces corneal parainflammation involving increased immune cell numbers after 24hours' (CD11c+, Lyz2+, γδ-T cells) and six days' (Ly6G+ cells) wear. We investigated the time course of onset and resolution of these responses. LysMcre or C57BL/6J mice were fitted with a contact lens (four to 48hours). Contralateral eyes did not wear lenses. After lens removal, Lyz2+, MHC-II+ or Ly6G+ cells were examined by quantitative imaging. RT-qPCR determined cytokine gene expression. Lens wear for 24hours increased corneal Lyz2+ cells versus contralateral eyes approximately two-fold. Corneas remained free of visible pathology. The Lyz2+ response was not observed after four or 12 hours' wear, nor after 12 hours' wear plus 12 hours' no wear. Lens removal after 24hours' wear further increased Lyz2+ cells (∼48% after one day), which persisted for four days, returning to baseline by seven days. Lyz2+ cells in contralateral eyes remained at baseline. MHC-II+ cells showed a similar response but without increasing after lens removal. Lens wear for 48 hours showed reduced Lyz2+ cells versus 24hours' wear with one day discontinuation, correlating with reduced IL-1β and IL-18 gene expression. Lens wear for 24hours did not induce Ly6G+ responses six days after removal. Lens-induced corneal parainflammation involving Lyz2+ cells requires 24hours' wear but persists after lens discontinuation, requiring seven days for reversal. Lens wear for 48 hours may suppress initial Lyz2+ cell and cytokine responses. The significance of parainflammation during and after lens wear remains to be determined.

Acetaminophen overdose causes a breach of the blood-bile barrier in mice but not in rats.

Acetaminophen (APAP) is known to cause a breach of the blood-bile barrier in mice that, via a mechanism called futile bile acid (BA) cycling, increases BA concentrations in hepatocytes above cytotoxic thresholds. Here, we compared this mechanism in mice and rats, because both species differ massively in their susceptibility to APAP and compared the results to available human data. Dose and time-dependent APAP experiments were performed in male C57BL6/N mice and Wistar rats. The time course of BA concentrations in liver tissue and in blood was analyzed by MALDI-MSI and LC-MS/MS. APAP and its derivatives were measured in the blood by LC-MS. APAP-induced liver damage was analyzed by histopathology, immunohistochemistry, and by clinical chemistry. In mice, a transient increase of BA in blood and in peri-central hepatocytes preceded hepatocyte death. The BA increase coincided with oxidative stress in liver tissue and a compromised morphology of bile canaliculi and immunohistochemically visualized tight junction proteins. Rats showed a reduced metabolic activation of APAP compared to mice. However, even at very high doses that caused cell death of hepatocytes, no increase of BA concentrations was observed neither in liver tissue nor in the blood. Correspondingly, no oxidative stress was detectable, and the morphology of bile canaliculi and tight junction proteins remained unaltered. In conclusion, different mechanisms cause cell death in rats and mice, whereby oxidative stress and a breach of the blood-bile barrier are seen only in mice. Since transient cholestasis also occurs in human patients with APAP overdose, mice are a clinically relevant species to study APAP hepatotoxicity but not rats.