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Trimethyltin chloride (TMT), an organotin compound with potent neurotoxicity, is widely used as a heat stabilizer for plastics. However, the precise pathogenic mechanism of TMT remains incompletely elucidated, and there persists a dearth of sensitive detection methodologies for early diagnosis of TMT. In this study, Sprague-Dawley rats were treated with 10 mg/kg TMT to simulate acute exposure in humans. Micro-positron emission tomography/computed tomography (Micro-PET/CT) and molecular imaging quantitative analysis tools were employed to calculate the uptake rate of 18 F-2-fluoro-D-deoxy-glucose in each functional region of brain tissue. At the same time, the neurobehavioral test and neuropathological results of the experimental rats were compared, aiming to assess the feasibility of PET/CT in the detection and localization of TMT nervous system damage from many aspects. The results showed that TMT decreased glucose uptake in a wide range of brain tissues in rats, and impaired the memory, muscle strength, coordination ability and emotion of rats. Moreover, TMT induced neuronal damage within the cerebral cortex, hippocampal CA1, CA3 and DG regions as well as the cerebellum while also promoting gliosis surrounding the hippocampus. PET/CT imaging results are highly consistent with behavioral and pathological results. In conclusion, TMT induces a widespread reduction in energy metabolism across various brain regions, and PET/CT can serve as a sensitive detection method for TMT-induced encephalopathy., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Extracellular vesicles (EVs) are small membrane-bound structures that play important roles in intercellular communication and the transfer of biomolecules between cells. EVs have become a topic of interest for research in translational proteomics for disease biomarker discovery due to their ability to reflect changes in the cellular proteome, including diseases affecting the brain. Utilizing the proteome analysis of EVs to its fullest potential requires proper isolation and purity. In this chapter, we describe a detailed method for the isolation and identification of brain tissue EVs for translational proteomics using our in-house chemical affinity magnetic bead-based (non-antibody) method, the EVtrap. We also discuss various methods for quantification, characterization, and functional analysis of isolated brain tissue EVs, including western blotting, and proteomic profiling of post-translational modifications (PTMs) involved in neurodegenerative diseases, such as protein N-terminal acetylation. This protocol provides a valuable resource for studies conducted on brain tissue EVs and their potential as biomarkers and therapeutic targets for neurological diseases., (© 2025. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

A surface-enhanced Raman scattering (SERS) biosensor with minimal invasiveness and high spatial resolution has been developed as a nanoendoscope to detect changes in protein concentrations at specific sites in biological tissues. While generally applicable to various tissues or proteins, the SERS nanoendoscope is demonstrated for the quantitative detection of S100β, an astrocytic protein whose plasmatic levels are known to vary in several neuropathologies such as Alzheimer's disease, schizophrenia, Down syndrome, Parkinson's disease and epilepsy, but for which intratissular levels have not been locally monitored, demonstrating key attributes of the SERS nanoendoscope. The SERS nanoendoscope is fabricated with densely and well-dispersed deposited gold nanoparticles modified with anti-S100β primary antibody on pulled optical fibers with a tip diameter of 700 nm, conducive to noninvasive and regiospecific detection of the S100β protein in different regions of mouse brain slices under different physiological stimuli with micrometer resolution. Quantification was performed ex vivo using SERS-active nanotags with secondary antibodies with detection limits of 5 and 7 nM in phosphate-buffered saline solution and mouse brain slice, respectively. Various physiological stimuli were then applied ex vivo to wild-type and S100β-knockout mouse brain slices to demonstrate the SERS nanoendoscope under physiological conditions. The average concentration of S100β was increased to 27, 45, and 48 nM upon N -methyl-d-aspartate, electrical, and optogenetic stimulation, respectively, statistically higher than all controls, demonstrating the ability of the SERS nanoendoscope to quantify protein release in biological tissues.

Background: Oleuropein (OLE) has the potential to reduce oxidative stress and inflammation. So, in the present investigation, we explored the protective effect of OLE on brain aging induced by d-galactose (D-Gal) in a rat model., Methods and Results: 40 Wister male adult rats were categorized into 5 groups. Group 1 received normal saline; group 2 was given 100 mg/kg of D-Gal intraperitoneally (IP). The rats in groups 3 to 5 were given D-Gal (100 mg/kg, IP) along with different doses of OLE (20, 40, and 80 mg/kg, respectively) orally. All administrations were performed daily for 8 weeks. 24 h after last treatment motor activity and memory impairment were evaluated. Then, the rats were euthanized and brain samples were collected for evaluating the levels of malondialdehyde (MDA), Brain-Derived Neurotrophic Factor (BDNF), protein carbonyl (PC), glutathione (GSH), glutathione peroxidase (GPX), catalase (CAT), Superoxide dismutase (SOD), Tumor necrosis factor alpha (TNF-α), interleukin 1 beta ( IL-1β), as well as Sirtuin 1 (SIRT1) and peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC1) gene expression. The results showed that D-Gal significantly reduced motor activity and memory performance (P < 0.05). It also significantly reduced the GPX, CAT and SOD activities, GSH and BDNF levels as well as SIRT1 and PGC1 expression, and, significantly increased PC, MDA TNF-α and IL-1β levels in the brain tissue (P < 0.05). Administration of OLE restored all of the above parameters close to control group., Conclusion: The findings demonstrated that OLE, through its antioxidant and anti-inflammatory properties, improved motor activity, memory impairment, and age-related neurological dysfunction., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Our brain undergoes significant micro- and macroscopic changes throughout its life cycle. It is therefore crucial to understand the effect of aging on the mechanical properties of the brain in order to develop accurate personalized simulations and diagnostic tools. Here we systematically probed the mechanical behavior of n=439 brain tissue samples in tension and compression, in different anatomical regions, for different axon orientations, across five age groups. We used Bayesian statistics to characterize the relation between brain age and mechanical properties and quantify uncertainties. Our results, based on our experimental data and material parameters for the isotropic Ogden and the anisotropic Gasser-Ogden-Holzapfel models, reveal a non-linear relationship between age and mechanics across the life cycle of the porcine brain. Both tensile and compressive shear moduli reached peak values ranging from 0.4-1.0 kPa in tension to 0.16-0.32 kPa in compression at three years of age. Anisotropy was most pronounced at six months, and then decreased. These results represent an important step in understanding age-dependent changes in the mechanical properties of brain tissue and provide the scientific basis for more accurate and realistic computational brain simulations. STATEMENT OF SIGNIFICANCE: In this paper, we investigate the age-dependent mechanical properties of brain tissue based on different deformation modes, anatomical regions, and axon orientations. Hierarchical Bayesian modeling was used to identify isotropic and anisotropic material parameters. The study reveals a nonlinear relationship between shear modulus, degree of anisotropy, and tension-compression asymmetry over the life cycle of the brain. By demonstrating the non-linearity of these relationships, the study fills a significant knowledge gap in current research. This work is a fundamental step in accurately characterizing the complex relationship between brain aging and mechanical properties., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Cold plasma-activated water (PAW) is a novel technology that was recently used in biomedical research; Despite its potential, PAW's safety remains inadequately assessed. The study explores the impact of PAW on behavioral responses and brain tissue histopathology in mice. Ten-week-old female albino mice were divided into three groups each containing 10 mice (5 replicates, 2 mice/cage) and received either distilled water (DW), or distilled water exposed to cold atmospheric plasma (CAP) for 3 min (PAW-3), or 15 min (PAW-15) by oral gavage in a dose of 200 μL/mice (3 times/week) for four weeks. PAW exhibited altered physicochemical properties compared to DW. Mice exposed to PAW demonstrated reduced burrowing activity, marble burying ability, and novel object recognition compared to controls, indicating potential neurobehavioral alterations. PAW-treated groups displayed notable histological lesions in brain tissues, including nerve cell necrosis, vascular congestion, and Purkinje cell degeneration, confirming neurotoxic effects. Positive reactions for NF-κB and iNOS in brain tissues of PAW-treated mice corroborated the histopathological findings, suggesting neuroinflammation and oxidative stress. The study highlights the need for further investigation into PAW's safety profile and optimal treatment protocols to mitigate potential neurobehavioral toxicity in biomedical research., Competing Interests: Declaration of Competing Interest The authors declare that there is no conflict of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

&lt;b&gt;Background and Objective:&lt;/b&gt; Nicotine-relevant smoking causes many serious issues of environmental pollution and complicated harm to human health. The present study aimed to evaluate the experimental effects of exposure to nicotine on the gene expression profiles of rat brain tissues with differentially expressed genes (DEGs). &lt;b&gt;Materials and Methods:&lt;/b&gt; The rat gene expression profiles of environmental exposure to nicotine were initially screened and retrieved from the microarray dataset GSE59895 in the GEO database. Next, it was analyzed with an integrated bioinformatics pipeline. The DEGs were analyzed in Limma and functional enrichment analyses of GO terms and KEGG pathways were performed with clusterProfiler. The STRING online tools and Cytoscape StringApp were subsequently employed to construct the protein-protein interaction (PPI) network, whereas key modules and hub genes were finally explored and visualized. &lt;b&gt;Results:&lt;/b&gt; There was total of 382 shared DEGs between different case groups in the experiment, whereas 9 common shared DEGs were found among all three groups. The significant enrichments of 28 GO terms and 3 KEGG pathways were comprehensively analyzed with corresponding functionally enriched genes. Then, 3 key modules and 10 hub genes were further identified and explored in the resulted PPI network. In the disease-related signaling pathways, eleven potential neuropathic disease-related genes may complement the treatment of neurodegenerative diseases. &lt;b&gt;Conclusion:&lt;/b&gt; The study found that chronic exposure to nicotine would result in the differential expression of the disease-related genes, whereas these DEGs might increase the environmental risks of Huntington's disease, Alzheimer's disease and other multiple neurodegenerative diseases.

The debate surrounding nature versus nurture remains a central question in neuroscience, psychology, and in psychiatry, holding implications for both aging processes and the etiology of mental illness. Epigenetics can serve as a bridge between genetic predisposition and environmental influences, thus offering a potential avenue for addressing these questions. Epigenetic clocks, in particular, offer a theoretical framework for measuring biological age based on DNA methylation signatures, enabling the identification of disparities between biological and chronological age. This structured review seeks to consolidate current knowledge regarding the relationship between mental disorders and epigenetic age within the brain. Through a comprehensive literature search encompassing databases such as EBSCO, PubMed, and ClinicalTrials.gov, relevant studies were identified and analyzed. Studies that met inclusion criteria were scrutinized, focusing on those with large sample sizes, analyses of both brain tissue and blood samples, investigation of frontal cortex markers, and a specific emphasis on schizophrenia and depressive disorders. Our review revealed a paucity of significant findings, yet notable insights emerged from studies meeting specific criteria. Studies characterized by extensive sample sizes, analysis of brain tissue and blood samples, assessment of frontal cortex markers, and a focus on schizophrenia and depressive disorders yielded particularly noteworthy results. Despite the limited number of significant findings, these studies shed light on the complex interplay between epigenetic aging and mental illness. While the current body of literature on epigenetic aging in mental disorders presents limited significant findings, it underscores the importance of further research in this area. Future studies should prioritize large sample sizes, comprehensive analyses of brain tissue and blood samples, exploration of specific brain regions such as the frontal cortex, and a focus on key mental disorders. Such endeavors will contribute to a deeper understanding of the relationship between epigenetic aging and mental illness, potentially informing novel diagnostic and therapeutic approaches., (© 2024. The Author(s).)

Significance: Mueller matrix imaging (MMI) is a comprehensive form of polarization imaging useful for assessing structural changes. However, there is limited literature on the polarimetric properties of brain specimens, especially with multispectral analysis., Aim: We aim to employ multispectral MMI for an exhaustive polarimetric analysis of brain structures, providing a reference dataset for future studies and enhancing the understanding of brain anatomy for clinicians and researchers., Approach: A multispectral wide-field MMI system was used to measure six fresh lamb brain specimens. Multiple decomposition methods (forward polar, symmetric, and differential) and polarization invariants (indices of polarimetric purity and anisotropy coefficients) have been calculated to obtain a complete polarimetric description of the samples. A total of 16 labels based on major brain structures, including grey matter (GM) and white matter (WM), were identified. K -nearest neighbors classification was used to distinguish between GM and WM and validate the feasibility of MMI for WM identification., Results: As the wavelength increases, both depolarization and retardance increase, suggesting enhanced tissue penetration into deeper layers. Moreover, utilizing multiple wavelengths allowed us to track dynamic shifts in the optical axis of retardance within the brain tissue, providing insights into morphological changes in WM beneath the cortical surface. The use of multispectral data for classification outperformed all results obtained with single-wavelength data and provided over 95% accuracy for the test dataset., Conclusions: The consistency of these observations highlights the potential of multispectral wide-field MMI as a non-invasive and effective technique for investigating the brain's architecture., (© 2024 The Authors.)

The continuous softening behavior of the brain tissue, i.e., the softening in the primary loading path with an increase in deformation, is modeled in this work as a state of hyperelasticity up to the onset of failure. That is, the softening behavior is captured via a core hyperelastic model without the addition of damage variables and/or functions. Examples of the application of the model will be provided to extant datasets of uniaxial tension and simple shear deformations, demonstrating the capability of the model to capture the whole-range deformation of the brain tissue specimens, including their softening behavior. Quantitative and qualitative comparisons with other models within the brain biomechanics literature will also be presented, showing the clear advantages of the current approach. The application of the model is then extended to capturing the rate-dependent softening behavior of the tissue by allowing the parameters of the core hyperelastic model to evolve, i.e., vary, with the deformation rate. It is shown that the model captures the rate-dependent and softening behaviors of the specimens favorably and also predicts the behavior at other rates. These results offer a clear set of advantages in favor of the considered modeling approach here for capturing the quasi-static and rate-dependent mechanical properties of the brain tissue, including its softening behavior, over the existing models in the literature, which at best may purport to capture only a reduced set of the foregoing behaviors, and with ill-posed effects., (Copyright © 2024 by ASME.)

The effect of strain rate and temperature on the hyperelastic material stress-strain characteristics of the damaged porcine brain tissue is evaluated in this present work. The desired constitutive responses are obtained using the commercially available finite element (FE) tool ABAQUS, utilizing 8-noded brick elements. The model's accuracy has been verified by comparing the results from the previously published literature. Further, the stress-strain behavior of the brain tissue is evaluated by varying the damages at various strain rates and temperatures (13, 20, 27, and 37°C) under compression test. Additionally, the sensitivity analysis of the model is computed to check the effect of input parameters, that is, the temperature, strain rate, and damages on the material properties (shear modulus). The modeling and discussion sections enumerate the inclusive features and model capabilities., (© 2024 Wiley Periodicals LLC.)

Purpose: To investigate the effect of inhaled oxygen level on dynamic glucose enhanced (DGE) MRI in mouse brain tissue and CSF at 3 T., Methods: DGE data of brain tissue and CSF from mice under normoxia or hyperoxia were acquired in independent and interleaved experiments using on-resonance variable delay multi-pulse (onVDMP) MRI. A bolus of 0.15 mL filtered 50% D-glucose was injected through the tail vein over 1 min during DGE acquisition. MRS was acquired before and after DGE experiments to confirm the presence of D-glucose., Results: A significantly higher DGE effect under normoxia than under hyperoxia was observed in brain tissue (p = 0.0001 and p = 0.0002 for independent and interleaved experiments, respectively), but not in CSF (p > 0.3). This difference is attributed to the increased baseline MR tissue signal under hyperoxia induced by a shortened T 1 and an increased BOLD effect. When switching from hyperoxia to normoxia without glucose injection, a signal change of ˜3.0% was found in brain tissue and a signal change of ˜1.5% was found in CSF., Conclusions: DGE signal was significantly lower under hyperoxia than that under normoxia in brain tissue, but not in CSF. The reason is that DGE effect size of brain tissue is affected by the baseline signal, which could be influenced by T 1 change and BOLD effect. Therefore, DGE experiments in which the oxygenation level is changed from baseline need to be interpreted carefully., (© 2024 The Authors. Magnetic Resonance in Medicine published by Wiley Periodicals LLC on behalf of International Society for Magnetic Resonance in Medicine.)

Objective: To investigate the development and dynamic changes of cysts in the brain of mice following infection with different forms of Toxoplasma gondii , so as to provide insights into for toxoplasmosis prevention and control., Methods: ICR mice at ages of 6 to 8 weeks, each weighing 20 to 25 g, were intraperitoneally injected with tachyzoites of the T. gondii PRU strain at a dose of 1 × 10 5 tachyzoites per mouse, orally administered with cysts at a dose of 20 oocysts per mouse or oocysts at a dose of 200 oocysts per mouse for modeling chronic T. gondii infection in mice, and the clinical symptoms and survival of mice were observed post-infection. Mice were orally infected with T. gondii cysts at doses of 10 (low-dose group), 20 (medium-dose group), 40 cysts per mouse (high-dose group), and the effect of different doses of T. gondii infections on the number of cysts was examined in the mouse brain. Mice were orally administered with T. gondii cysts at a dose of 20 cysts per mouse, and grouped according to gender (female and male) and time points of infections (20, 30, 60, 90, 120, 150, 180 days post-infection), and the effects of gender and time points of infections on the number of cysts was examined in the mouse brain. In addition, mice were divided into the tachyzoite group (Group T), the first-generation cyst group (Group C1), the second-generation cyst group (Group C2), the third-generation cyst (Group C3) and the fourth-generation cyst group (Group C4). Mice in the Group T were intraperitoneally injected with T. gondii tachyzoites at a dose of 1 × 10 5 tachyzoites per mouse, and the cysts were collected from the mouse brain tissues 30 days post-infection, while mice in the Group C1 were orally infected with the collected cysts at a dose of 30 cysts per mouse. Continuous passage was performed by oral administration with cysts produced by the previous generation in mice, and the effect of continuous passage on the number of cysts was examined in the mouse brain., Results: Following infection with T. gondii tachyzoites, cysts and oocysts in mice, obvious clinical symptoms were observed on days 6 to 13 and mice frequently died on days 7 to 12. The survival rates of mice were 67.0%, 87.0% and 53.0%, and the mean numbers of cysts were (516.0 ± 257.2), (1 203.0 ± 502.0) and (581.0 ± 183.1) in the mouse brain ( F = 11.94, P < 0.01) on day 30 post-infection with T. gondii tachyzoites, cysts and oocysts, respectively, and the numbers of cysts in the brain tissues were significantly lower in mice infected with T. gondii tachyzoites and oocysts than in those infected with cysts (all P values < 0.01). The survival rates of mice were 87.0%, 87.0% and 60.0%, and the mean numbers of cysts were (953.0 ± 355.5), (1 084.0 ± 474.3) and (1 113.0 ± 546.0) in the mouse brain in the low-, medium- and high-dose groups on day 30 post-infection, respectively ( F = 0.42, P > 0.05). The survival rates of male and female mice were 73.0% and 80.0%, and the mean numbers of cysts were (946.4 ± 411.4) and (932.1 ± 322.4) in the brain tissues of male and female mice, respectively ( F = 1.63, P > 0.05). Following continuous passage, the mean numbers of cysts were (516.0 ± 257.2), (1 203.0 ± 502.0), (896.8 ± 332.3), (782.5 ± 423.9) and (829.2 ± 306.0) in the brain tissues of mice in the T, C1, C2, C3 and C4 groups, respectively ( F = 4.82, P < 0.01), and the number of cysts was higher in the mouse brain in Group 1 than in Group T ( P < 0.01). Following oral administration of 20 T. gondii cysts in mice, cysts were found in the moues brain for the first time on day 20 post-infection, and the number of cysts gradually increased over time, peaked on days 30 and 90 post-infection and then gradually decreased; however, the cysts were still found in the mouse brain on day 180 post-infection., Conclusions: There is a higher possibility of developing chronic T. gondii infection in mice following infection with cysts than with oocysts or tachyzoites and the most severe chronic infection is seen following infection with cysts. The number of cysts does not correlate with the severity of chronic T. gondii infection, and the number of cysts peaks in the mouse brain on days 30 and 90 post-infection.

A new modelling approach is employed in this work for application to the rate-dependent mechanical behaviour of the brain tissue, as an incompressible isotropic material. Extant datasets encompassing single- and multi-mode compression, tension and simple shear deformation(s) are considered, across a wide range of deformation rates from quasi-static to rates akin to blast loading conditions, in the order of 1000 s -1 . With a simple functional form and a reduced number of parameters, the model is shown to capture the considered rate-dependent behaviours favourably, including in both single- and multi-mode deformation fits, and over all range of deformation rates. The provided modelling results here are obtained from either first fitting the model to the quasi-static data, or/and predicting the behaviour at a different rate than those used for calibrating the model parameters. Given its simplicity, versatility, predictive capability and accuracy, the application of the utilised modelling framework in this work to the rate-dependent mechanical behaviour of the brain tissue is proposed., Competing Interests: Declaration of competing interest The authors have no competing interests to declare., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Finite element head models offer great potential to study brain-related injuries; however, at present may be limited by geometric and material property simplifications required for continuum-level human body models. Specifically, the mechanical properties of the brain tissues are often represented with simplified linear viscoelastic models, or the material properties have been optimized to specific impact cases. In addition, anatomical structures such as the arachnoid complex have been omitted or implemented in a simple lumped manner. Recent material test data for four brain regions at three strain rates in three modes of loading (tension, compression, and shear) was used to fit material parameters for a hyper-viscoelastic constitutive model. The material model was implemented in a contemporary detailed head finite element model. A detailed representation of the arachnoid trabeculae was implemented with mechanical properties based on experimental data. The enhanced head model was assessed by re-creating 11 ex vivo head impact scenarios and comparing the simulation results with experimental data. The hyper-viscoelastic model faithfully captured mechanical properties of the brain tissue in three modes of loading and multiple strain rates. The enhanced head model showed a high level of biofidelity in all re-created impacts in part due to the improved brain-skull interface associated with implementation of the arachnoid trabeculae. The enhanced head model provides an improved predictive capability with material properties based on tissue level data and is positioned to investigate head injury and tissue damage in the future., (© 2024. The Author(s) under exclusive licence to Biomedical Engineering Society.)

Pulsed electric fields are extensively utilized in clinical treatments, such as subthalamic deep brain stimulation, where electric field loading is in direct contact with brain tissue. However, the alterations in brain tissue's mechanical properties and microstructure due to changes in electric field parameters have not received adequate attention. In this study, the mechanical properties and microstructure of the brain tissue under pulsed electric fields were focused on. Herein, a custom indentation device was equipped with a module for electric field loading. Parameters such as pulse amplitude and frequency were adjusted. The results demonstrated that following an indentation process lasting 5 s and reaching a depth of 1000 μm, and a relaxation process of 175 s, the average shear modulus of brain tissue was reduced, and viscosity decreased. At the same amplitude, high-frequency pulsed electric fields had a smaller effect on brain tissue than low-frequency ones. Furthermore, pulsed electric fields induced cell polarization and reduced the proteoglycan concentration in brain tissue. As pulse frequency increased, cell polarization diminished, and proteoglycan concentration decreased significantly. High-frequency pulsed electric fields applied to brain tissue were found to reduce impedance fluctuation amplitude. This study revealed the effect of pulsed electric fields on the mechanical properties and microstructure of ex vivo brain tissue, providing essential information to promote the advancement of brain tissue electrotherapy in clinical settings., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

The release of metabolites from their bound to free forms is the main regulatory path in living species. Therefore, the ability to determine the free concentrations of small molecules is highly critical in many biological samples. The main challenges in achieving this task are the interferences inherent to complex matrices and the ability to distinguish between the free and total concentrations. This paper presents a non-invasive microextraction method that enables the determination of endocannabinoids in brain tissue. The proposed method is based on two key principles: the availability of the free concentration of endocannabinoids for partitioning to the solid-phase microextraction (SPME) fiber; and negligible depletion enabled by the small volume of extraction phase on the fiber. These features allow the presented SPME method to provide information about the free concentration of analytes without disturbing the binding equilibrium between the analytes and the matrix. The determination of spiked samples with known concentrations enables the percentage of analyte bound to the tissue to be calculated, which can then be applied to calculate the total concentration from the determined free concentration. This manuscript focuses on the determination of the free concentration and tissue binding percentages of endocannabinoids in brain tissue. Significantly, SPME's small size and potential for non-invasive sampling enable its application in live animal subjects with minimal tissue damage., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Electrotherapy plays a crucial role in regulating neuronal activity. Nevertheless, the relevant therapeutic mechanisms are still unclear; thus, the effects of electric fields on brain tissue's mechanical properties and microstructure need to be explored. In this study, focusing on the changes in mechanical properties and microstructure of ex vivo porcine brain tissues under different types of electric fields, directional and alternating electric fields (frequencies of 5, 20, 50, and 80 Hz, respectively) integrate with a custom-designed indentation device. The experimental results showed that for the ex vivo brain tissue, the directional electric field (DEF) can reduce the elastic properties of brain tissue. Simultaneously, the DEF can increase the cell spacing and reduce the proteoglycan content. The transmission electron microscope (TEM) analysis observed that the DEF can reduce the integrity of the plasma membrane, the endoplasmic reticulum's stress response, and the myelin lamella's separation. The alternating electric field (AEF) can accelerate the stress relaxation process of brain tissue and change the time-dependent mechanical properties of brain tissue. Meanwhile, with the increase in frequency, the cell spacing decreased, and the proteoglycan content gradually approached the control group without electric fields. TEM analysis observed that with the increase in frequency, the integrity of the plasma membrane increases, and the separation of the myelin lamella gradually disappears. Understanding the changes in the mechanical properties and microstructure of brain tissue under AEF and DEF enables a preliminary exploration of the therapeutic mechanism of electrotherapy. Simultaneously, the essential data was provided to support the development of embedded electrodes. In addition, the ex vivo experiments build a solid foundation for future in vivo experiments.

Two-dimensional mesoscale finite element analysis (FEA) of a multilayered brain tissue was performed to calculate the damage-related average stress triaxiality and local maximum von Mises strain in the brain. The FEA was integrated with rate-dependent hyperelastic and internal state variable (ISV) models, respectively, describing the behaviors of wet and dry brain tissues. Using the finite element results, a statistical method of design of experiments (DOE) was utilized to independently screen the relative influences of seven parameters related to brain morphology (sulcal width/depth, gray matter (GM) thickness, cerebrospinal fluid (CSF) thickness and brain lobe) and loading/environment conditions (strain rate and humidity) with respect to the potential damage growth/coalescence in the brain tissue. The results of the parametric study illustrated that the GM thickness and humidity were the two most crucial parameters affecting average stress triaxiality. For the local maximum von Mises strain at the depth of brain sulci, the brain lobe/region was the most influential factor. The conclusion of this investigation gives insight for the future development and refinement of a macroscale brain damage model incorporating information from lower length scale., (Copyright © 2022 by ASME.)

This study focuses on the variations in the brain tissue dynamic behaviour pointing out new insight into the material nonlinear viscoelasticity. Shear dynamic response curves are obtained in different working conditions in terms of strain sweep and superimposed static compression offsets (SCO) applied in orthogonal direction to the shear. The strain sweep mode is used to study the storage and loss moduli dependence on the amplitude of the applied strain. It is found that the material exhibits linear viscoelastic behaviour up to about 0.1% strain amplitude. Above this critical threshold, the storage modulus G' decreases rapidly with increasing dynamic strain amplitude and this effect is gradually intensified as the SCO are increased. In addition, it is observed that the loss factor (G''/G') increases by increasing the SCO applied to the specimens. The dynamic strain amplitude results of the storage modulus reveal that the elastic component of the brain tissue's stiffness (G') evaluated at low strain strongly increases with increasing static superimposed compression strain while the loss factor in the same strain range appears to be SCO independent. Finally, dynamic stiffness recovery after a large strain deformation is considered. The reduction in low amplitude dynamic modulus and subsequent recovery kinetics due to a perturbation is found to be independent of the level of the SCO. The same assessments were carried out on 5 consecutive strain sweep cycle loading. It has been noticed that at the last cycle, the dissipation peak is reduced, and the non-linearity of the curve begins earlier. This could be explained by the effects of cerebral edema on cells and their surrounding environment., (Copyright © 2022. Published by Elsevier Ltd.)

Brain tissue is one of the most complex and softest tissues in the human body. Due to its ultrasoft and biphasic nature, it is difficult to control the deformation state during biomechanical testing and to quantify the highly nonlinear, time-dependent tissue response. In numerous experimental studies that have investigated the mechanical properties of brain tissue over the last decades, stiffness values have varied significantly. One reason for the observed discrepancies is the lack of standardized testing protocols and corresponding data analyses. The tissue properties have been tested on different length and time scales depending on the testing technique, and the corresponding data have been analyzed based on simplifying assumptions. In this review, we highlight the advantage of using nonlinear continuum mechanics based modeling and finite element simulations to carefully design experimental setups and protocols as well as to comprehensively analyze the corresponding experimental data. We review testing techniques and protocols that have been used to calibrate material model parameters and discuss artifacts that might falsify the measured properties. The aim of this work is to provide standardized procedures to reliably quantify the mechanical properties of brain tissue and to more accurately calibrate appropriate constitutive models for computational simulations of brain development, injury and disease. Computational models can not only be used to predictively understand brain tissue behavior, but can also serve as valuable tools to assist diagnosis and treatment of diseases or to plan neurosurgical procedures. © 2022 The Authors. Current Protocols published by Wiley Periodicals LLC., (© 2022 The Authors. Current Protocols published by Wiley Periodicals LLC.)

Neurodegenerative movement disorders (NMDs) are age-dependent disorders that are characterised by the degeneration and loss of neurons, typically accompanied by pathological accumulation of different protein aggregates in the brain, which lead to motor symptoms. NMDs include Parkinson's disease, multiple system atrophy, progressive supranuclear palsy, and Huntington's disease, among others. Epigenetic modifications are responsible for functional gene regulation during development, adult life and ageing and have progressively been implicated in complex diseases such as cancer and more recently in neurodegenerative diseases, such as NMDs. DNA methylation is by far the most widely studied epigenetic modification and consists of the reversible addition of a methyl group to the DNA without changing the DNA sequence. Although this research field is still in its infancy in relation to NMDs, an increasing number of studies point towards a role for DNA methylation in disease processes. This review addresses recent advances in epigenetic and epigenomic research in NMDs, with a focus on human brain DNA methylation studies. We discuss the current understanding of the DNA methylation changes underlying these disorders, the potential for use of these DNA modifications in peripheral tissues as biomarkers in early disease detection, classification and progression as well as a promising role in future disease management and therapy., (© 2021 The Authors. Neuropathology and Applied Neurobiology published by John Wiley & Sons Ltd on behalf of British Neuropathological Society.)

Inorganic elements have been associated with brain tumours for long. The blood concentration of 47 elements was assessed by ICP-MS in 26 brain tumour patients and 21 healthy subjects from Bucharest (Romania). All 47 elements were detected in the brain tumour tissue, and 22 were detected in > 80% of samples; this implies that these elements can cross the blood-brain barrier. Median blood levels of cadmium, lead, and nickel were higher than the reference values (1.14, 53.3, and 2.53 ng/mL). Gadolinium and tantalum showed significantly higher concentrations among cases. We observed considerable differences and different profiles of the presence of inorganic elements between the tumour and non-tumour brain tissue and between tissue from the primary tumour and tissue from brain metastasis. Our data suggest that similar to heavy metals, other elements - commonly used in high tech devices and rare earth elements - can also influence brain tumour.

Interkingdom communication between bacteria and host organisms is one of the most interesting research topics in biology. Quorum sensing molecules produced by Gram-negative bacteria, such as acylated homoserine lactones and quinolones, have been shown to interact with host cell receptors, stimulating innate immunity and bacterial clearance. To our knowledge, there is no evidence that these molecules influence CNS function. Here, we have found that low micromolar concentrations of the Pseudomonas aeruginosa quorum sensing autoinducer, 2-heptyl-3-hydroxy-4-quinolone (PQS), inhibited polyphosphoinositide hydrolysis in mouse brain slices, whereas four selected acylated homoserine lactones were inactive. PQS also inhibited forskolin-stimulated cAMP formation in brain slices. We therefore focused on PQS in our study. Biochemical effects of PQS were not mediated by the bitter taste receptors, T2R4 and T2R16. Interestingly, submicromolar concentrations of PQS could be detected in the serum and brain tissue of adult mice under normal conditions. Levels increased in five selected brain regions after single i.p. injection of PQS (10 mg/kg), peaked after 15 min, and returned back to normal between 1 and 4 h. Systemically administered PQS reduced spontaneous locomotor activity, increased the immobility time in the forced swim test, and largely attenuated motor response to the psychostimulant, methamphetamine. These findings offer the first demonstration that a quorum sensing molecule specifically produced by Pseudomonas aeruginosa is centrally active and influences cell signaling and behavior. Quorum sensing autoinducers might represent new interkingdom signaling molecules between ecological communities of commensal, symbiotic, and pathogenic microorganisms and the host CNS., (Copyright © 2021. Published by Elsevier Ltd.)

Brain tissue is a heterogeneous material, constituted by a soft matrix filled with cerebrospinal fluid. The interactions between, and the complexity of each of these components are responsible for the non-linear rate-dependent behaviour that characterises what is one of the most complex tissue in nature. Here, we investigate the influence of the cutting rate on the fracture properties of brain, through wire cutting experiments. We also present a computational model for the rate-dependent behaviour of fracture propagation in soft materials, which comprises the effects of fluid interaction through a poro-hyperelastic formulation. The method is developed in the framework of finite strain continuum mechanics, implemented in a commercial finite element code, and applied to the case of an edge-crack remotely loaded by a controlled displacement. Experimental and numerical results both show a toughening effect with increasing rates, which is linked to the energy dissipated by the fluid-solid interactions in the region surrounding the crack tip., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

While the degree of cerebellar tonsillar descent is considered the primary radiologic marker of Chiari malformation type I (CMI), biomechanical forces acting on the brain tissue in CMI subjects are less studied and poorly understood. In this study, regional brain tissue displacement and principal strains in 43 CMI subjects and 25 controls were quantified using a magnetic resonance imaging (MRI) methodology known as displacement encoding with stimulated echoes (DENSE). Measurements from MRI were obtained for seven different brain regions-the brainstem, cerebellum, cingulate gyrus, corpus callosum, frontal lobe, occipital lobe, and parietal lobe. Mean displacements in the cerebellum and brainstem were found to be 106 and 64% higher, respectively, for CMI subjects than controls (p < .001). Mean compression and extension strains in the cerebellum were 52 and 50% higher, respectively, in CMI subjects (p < .001). Brainstem mean extension strain was 41% higher in CMI subjects (p < .001), but no significant difference in compression strain was observed. The other brain structures revealed no significant differences between CMI and controls. These findings demonstrate that brain tissue displacement and strain in the cerebellum and brainstem might represent two new biomarkers to distinguish between CMI subjects and controls.

(1) Recently, metabolic profiling of the tissue in the native state or extracts of its metabolites has become increasingly important in the field of metabolomics. An important factor, in this case, is the presence of blood in a tissue sample, which can potentially lead to a change in the concentration of tissue metabolites and, as a result, distortion of experimental data and their interpretation. (2) In this paper, the metabolomic profiling based on NMR spectroscopy was performed to determine the effect of blood contained in the studied samples of brain tissue on their metabolomic profile. We used 13 male laboratory CD-1 ® IGS mice for this study. The animals were divided into two groups. The first group of animals ( n = 7) was subjected to the perfusion procedure, and the second group of animals ( n = 6) was not perfused. The brain tissues of the animals were homogenized, and the metabolite fraction was extracted with a water/methanol/chloroform solution. Samples were studied by high-frequency 1 H-NMR spectroscopy with subsequent statistical data analysis. The group comparison was performed with the use of the Student's test. We identified 36 metabolites in the brain tissue with the use of NMR spectroscopy. (3) For the major set of studied metabolites, no significant differences were found in the brain tissue metabolite concentrations in the native state and after the blood removal procedure. (4) Thus, it was shown that the presence of blood does not have a significant effect on the metabolomic profile of the brain in animals without pathologies.

Jian carp (Cyprinus carpio var. Jian) is an economically important cultured fish in China. Currently, it is facing threats from infectious diseases including koi herpesvirus (KHV). Here, we established a new cell line, designated CCB-J, derived from the brain tissue of the Jian carp. CCB-J cells grew well in Leibovitz's L-15 medium containing 20% fetal bovine serum at 25 °C and have been subcultured for more than 60 passages. At the 30th passage, analysis showed that the number of chromosomes was 100, which is identical to that of other carp variants. Sequencing of the 18S ribosomal DNA confirmed that CCB-J originated from Jian carp. After transfection with the pEGFP-N1 plasmid, green fluorescence was observed in CCB-J. The replication of KHV in CCB-J cells was confirmed by RT-PCR and transmission electron microscopy. The viral titers of KHV in CCB-J cells and CCB cells, which have been widely used in the study of KHV, reached 10 3.9 and 10 1.8 median tissue culture infectious dose (TCID 50 /mL), respectively, within 14 days. The result of TaqMan PCR revealed that CCB-J cells were more sensitive to KHV than CCB cells. Meanwhile, a cytopathic effect (CPE) was also observed in the CCB-J cells in a shorter time post-infection compared with CCB cells. In summary, the CCB-J cell line will be a useful tool in the study of viral pathogenesis and vaccine research.

Gene expression studies are reported to be influenced by pre-analytical factors that can compromise RNA yield and integrity, which in turn may confound the experimental findings. Here we investigate the impact of four pre-analytical factors on brain-derived RNA: time-before-collection, tissue specimen size, tissue collection method, and RNA isolation method. We report no significant differences in RNA yield or integrity between 20 mg and 60 mg tissue samples collected in either liquid nitrogen or the RNAlater stabilizing solution. Isolation of RNA employing the TRIzol reagent resulted in a higher yield compared to isolation via the QIAcube kit while the latter resulted in RNA of slightly better integrity. Keeping brain tissue samples at room temperature for up to 160 min prior to collection and isolation of RNA resulted in no significant difference in yield or integrity. Our findings have significant practical and financial consequences for clinical genomic departments and other laboratory settings performing large-scale routine RNA expression analysis of brain samples.

Feed efficiency (FE) is one of the most important economic traits in the porcine industry. In this study, high-throughput RNA sequencing (RNA-seq) was first utilized for brain tissue transcriptome analysis in pigs to indicate the potential genes and biological pathways related to FE in pigs. A total of 8 pigs with either extremely high-FE group (HE-group) or low-FE group (LE-group) were selected from 225 Duroc × (Landrace × Yorkshire) (DLY) pigs for transcriptomic analysis. RNA-seq analysis was performed to determine differentially expressed genes (DEGs) between the HE- and LE-group, and 430 DEGs were identified in brain tissues of pigs (|log 2 (FoldChange)| > 1; adjusted p -values <0.05). Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis showed that the DEGs were mainly enriched in synaptic signaling or transmission, and hormone secretion pathways, in which insulin secretion, and oxytocin signaling pathways were closely associated with FE by regulating feeding behavior and energy metabolism (adjusted p -values <0.05). Further, the transcription factors (TFs) analysis and gene co-expression network analysis indicated three hub differentially expressed TFs ( NR2F2 , TFAP2D , and HNF1B ) that affected FE by mainly regulating feeding behavior, insulin sensitivity, or energy metabolism. Our findings suggest several potential TFs and biological pathways for further investigations of FE in pigs.

Trichlorfon is widely used to control pest insects and various parasitic infestations in agriculture, aquaculture and human medicine. However, the long-term widespread use and overuse of trichlorfon poses risks to public and environmental health. Thus, the aim of this study was to evaluate the interference of trichlorfon on gene transcription patterns in the brain of Rana chensinensis with 4 weeks treatment under control conditions and 0.1 mg/L exposure. In total, 102,013 unigenes were obtained from the brain tissue of R. chensinensis , and 874 differentially expressed genes (DEGs) were identified. Functional annotation indicated that out of 118,643 unigenes, 45,600 (44.7%) were annotated in the Nr, Nt, the Swiss-Prot, KEGG, COG, and GO databases. The differential expression patterns of 4 genes associated with neural activity were selected and validated by quantitative polymerase chain reaction (qPCR). The results revealed that except for the canonical cholinesterase-based mechanism, trichlorfon could act on other receptors and alter certain types of neuronal ion channels as the major target sites. All of these effects ultimately cause disorders of multifunctional pathways and other neurotransmitter pathways in the host. The results further our understanding of the mechanisms underlying nontarget effects of organophosphate insecticides (OPs) through multitargets studies.

There is growing evidence that mechanical factors affect brain functioning. However, brain components responsible for regulating the physiological mechanical environment are not completely understood. To determine the relationship between structure and stiffness of brain tissue, we performed high-resolution viscoelastic mapping by dynamic indentation of the hippocampus and the cerebellum of juvenile mice brains, and quantified relative area covered by neurons (NeuN-staining), axons (neurofilament NN18-staining), astrocytes (GFAP-staining), myelin (MBP-staining) and nuclei (Hoechst-staining) of juvenile and adult mouse brain slices. Results show that brain subregions have distinct viscoelastic parameters. In gray matter (GM) regions, the storage modulus correlates negatively with the relative area of nuclei and neurons, and positively with astrocytes. The storage modulus also correlates negatively with the relative area of myelin and axons (high cell density regions are excluded). Furthermore, adult brain regions are ∼ 20%-150% stiffer than the comparable juvenile regions which coincide with increase in astrocyte GFAP-staining. Several linear regression models are examined to predict the mechanical properties of the brain tissue based on (immuno)histochemical stainings., (Copyright © 2020 The Authors. Published by Elsevier Ltd.. All rights reserved.)

We describe five cases of fatally injured males (occupational accident, car driver, pedestrian, motorcyclist and suicidal jump from great height) with one universal autopsy finding - the presence of brain tissue in one or both auditory canals. Internal examination revealed that all victims had multiple head fractures with dura lacerations. In four cases, the petrous part of the temporal bone was fractured (hinge fracture), while in one case the fracture of both the petrous part of the temporal bones and the occipital bone (ring fracture) was present. In all of these cases, considerable pressure was applied to the head, pushing brain tissue equally in all directions (due to incompressibility of the tissue). The tissue followed the path of least resistance, going through the lacerated dura into the fractured petrous part of the temporal bones and finally reaching the middle ear cavity and auditory canal. This phenomenon is almost exclusively encountered in closed-head injuries. In an open-head injury, brain tissue would be expelled through the open bone fracture and scalp wound. The presence of brain tissue in the ears could indicate a hinge or ring fracture in a closed-head injury which occurred as the result of excessive impulse force or considerable pressure applied to the head, i.e. the head was compressed and/or squeezed.

Many factors contribute to the health risks encountered by astronauts on missions outside Earth's atmosphere. Spaceflight-induced potential adverse neurovascular damage and late neurodegeneration are a chief concern. The goal of the present study was to characterize the effects of spaceflight on oxidative damage in the mouse brain and its impact on blood-brain barrier (BBB) integrity. Ten-week-old male C57BL/6 mice were launched to the International Space Station (ISS) for 35 days as part of Space-X 12 mission. Ground control (GC) mice were maintained on Earth in flight hardware cages. Within 38 ± 4 hours after returning from the ISS, mice were euthanized and brain tissues were collected for analysis. Quantitative assessment of brain tissue demonstrated that spaceflight caused an up to 2.2-fold increase in apoptosis in the hippocampus compared to the control group. Immunohistochemical analysis of the mouse brain revealed an increased expression of aquaporin4 (AQP4) in the flight hippocampus compared to the controls. There was also a significant increase in the expression of platelet endothelial cell adhesion molecule-1 (PECAM-1) and a decrease in the expression of the BBB-related tight junction protein, Zonula occludens-1 (ZO-1). These results indicate a disturbance of BBB integrity. Quantitative proteomic analysis showed significant alterations in pathways responsible for neurovascular integrity, mitochondrial function, neuronal structure, protein/organelle transport, and metabolism in the brain after spaceflight. Changes in pathways associated with adhesion and molecular remodeling were also documented. These data indicate that long-term spaceflight may have pathological and functional consequences associated with neurovascular damage and late neurodegeneration., (© 2020 The Authors. The FASEB Journal published by Wiley Periodicals LLC on behalf of Federation of American Societies for Experimental Biology.)

The biophysical modeling efforts in diffusion MRI have grown considerably over the past 25 years. In this review, we dwell on the various challenges along the journey of bringing a biophysical model from initial design to clinical implementation, identifying both hurdles that have been already overcome and outstanding issues. First, we describe the critical initial task of selecting which features of tissue microstructure can be estimated using a model and which acquisition protocol needs to be implemented to make the estimation possible. The model performance should necessarily be tested in realistic numerical simulations and in experimental data - adapting the fitting strategy accordingly, and parameter estimates should be validated against complementary techniques, when/if available. Secondly, the model performance and validity should be explored in pathological conditions, and, if appropriate, dedicated models for pathology should be developed. We build on examples from tumors, ischemia and demyelinating diseases. We then discuss the challenges associated with clinical translation and added value. Finally, we single out four major unresolved challenges that are related to: the availability of a microstructural ground truth, the validation of model parameters which cannot be accessed with complementary techniques, the development of a generalized standard model for any brain region and pathology, and the seamless communication between different parties involved in the development and application of biophysical models of diffusion., Competing Interests: Declaration of Competing Interest IOJ is a co-inventor on U.S. Patent No. 10360472 relative to a “System, method and computer-accessible medium for determining brain microstructure parameters from diffusion magnetic resonance”. Other co-authors have no competing interests to declare., (Copyright © 2020 Elsevier B.V. All rights reserved.)

C1q/tumor necrosis factor (TNF)-related proteins (CTRPs) particularly CTRP9, have been established to be as adiponectin (APN) highly conserved paralogs which assemble several APN regulatory functions. Recently, growing body of evidences drawn significant attention to evaluate metabolic and cardiovascular effect of CTRP9. However, the potential role of CTRP9 in brain tissue has not yet fully illustrated. Here, we aimed to uncover latest advances regarding the CTRP9 related signaling pathways and during brain aging process., Competing Interests: Declaration of Competing Interest The authors declare that there is no conflict of interest., (Copyright © 2020. Published by Elsevier Inc.)

Tissue longitudinal relaxation characterized by recovery time T 1 or rate R 1 is a fundamental MRI contrast mechanism that is increasingly being used to study the brain's myelination patterns in both health and disease. Nevertheless, the quantitative relationship between T 1 and myelination, and its dependence on B 0 field strength, is still not well known. It has been theorized that in much of brain tissue, T 1 field-dependence is driven by that of macromolecular protons (MP) through a mechanism called magnetization transfer (MT). Despite the explanatory power of this theory and substantial support from in-vitro experiments at low fields (<3 ​T), in-vivo evidence across clinically relevant field strengths is lacking. In this study, T 1 -weighted MRI was acquired in a group of eight healthy volunteers at four clinically relevant field strengths (0.55, 1.5, 3 and 7 ​T) using the same pulse sequence at a single site, and jointly analyzed based on the two-pool model of MT. MP fraction and free-water pool T 1 were obtained in several brain structures at 3 and 7 ​T, which allowed distinguishing between contributions from macromolecular content and iron to tissue T 1 . Based on this, the T 1 of MP in white matter, indirectly determined by assuming a field independent T 1 of free water, was shown to increase approximately linearly with B 0 . This study advances our understanding of the T 1 contrast mechanism and its relation to brain myelin content across the wide range of currently available MRI strengths, and it has the potential to inform design of T 1 mapping methods for improved reproducibility in the human brain., (Published by Elsevier Inc.)

Extracellular vesicle (EV) is a unified terminology of membrane-enclosed vesicular species ubiquitously secreted by almost every cell type and present in all body fluids. They carry a cargo of lipids, metabolites, nucleic acids and proteins for their clearance from cells as well as for cell-to-cell communications. The exact composition of EVs and their specific functions are not well understood due to the underdevelopment of the separation protocols, especially those from the central nervous system including animal and human brain tissues as well as cerebrospinal fluids, and the low yield of proteins in the separated EVs. To understand their exact molecular composition and their functional roles, development of the reliable protocols for EV separation is necessary. Here we report the methods for EV separation from human and mouse unfixed frozen brain tissues by a sucrose step gradient ultracentrifugation method, and from human cerebrospinal fluids by an affinity capture method. The separated EVs were assessed for morphological, biophysical and proteomic properties of separated EVs by nanoparticle tracking analysis, transmission electron microscopy, and labeled and label-free mass spectrometry for protein profiling with step-by-step protocols for each assessment., (Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.)

In this paper, the dynamic behavior of bovine brain tissue, measured from a set of in vitro experiments, is investigated and represented through a nonlinear viscoelastic constitutive model. The brain samples were tested by employing unconfined compression tests at three different deformation rates of 10, 100, and 1000 mm/s. The tissue exhibited a significant rate-dependent behavior with different compression speeds. Based on the parallel rheological framework approach, a nonlinear viscoelastic model that captures the key aspects of the rate dependency in large-strain behavior was introduced. The proposed model was numerically calibrated to the tissue test data from three different deformation rates. The determined material parameters provided an excellent constitutive representation of tissue response in comparison with the test results. The obtained material parameters were employed in finite element simulations of tissue under compression loadings and successfully verified by the experimental results, thus demonstrating the computational compatibility of the proposed material model. The results of this paper provide groundwork in developing a characterization framework for large-strain and rate-dependent behavior of brain tissue at moderate to high strain rates which is of the highest importance in biomechanical analysis of the traumatic brain injury.

Our previous study revealed a higher incidence of gene dynamic mutation in newborns conceived by IVF, highlighting that IVF may be disruptive to the DNA stability of IVF offspring. However, the underlying mechanisms remain unclear. The DNA damage repair system plays an essential role in gene dynamic mutation and neurodegenerative disease. To evaluate the long-term impact of IVF on DNA damage repair genes, we established an IVF mouse model and analyzed gene and protein expression levels of MSH2, MSH3, MSH6, MLH1, PMS2, OGG1, APEX1, XPA and RPA1 and also the amount of H2AX phosphorylation of serine 139 which is highly suggestive of DNA double-strand break (γH2AX expression level) in the brain tissue of IVF conceived mice and their DNA methylation status using quantitative real-time PCR, western blotting and pyrosequencing. Furthermore, we assessed the capacity of two specific non-physiological factors in IVF procedures during preimplantation development. The results demonstrated that the expression and methylation levels of some DNA damage repair genes in the brain tissue of IVF mice were significantly changed at 3 weeks, 10 weeks and 1.5 years of age, when compared with the in vivo control group. In support of mouse model findings, oxygen concentration of in vitro culture environment was shown to have the capacity to modulate gene expression and DNA methylation levels of some DNA damage repair genes. In summary, our study indicated that IVF could bring about long-term alterations of gene and protein expression and DNA methylation levels of some DNA damage repair genes in the brain tissue and these alterations might be resulted from the different oxygen concentration of culture environment, providing valuable perspectives to improve the safety and efficiency of IVF at early embryonic stage and also throughout different life stages., (© The Author(s) 2020. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Tizoxanide, the active metabolite of nitazoxanide, has recently been reported as an effective agent for the treatment of glioma. As there had been no report about the analysis of tizoxanide in brain tissue, we established extraction and UHPLC-MS/MS methods to quantify tizoxanide in rat brain and plasma to evaluate the brain-to-plasma ratio of tizoxanide. The biological samples were mainly prepared by acetonitrile and the separation was performed on a Waters XBridge® BEH C 18 column. The mobile phase was composed of water mixed with 10 mm ammonium formate (pH 3.0) and acetonitrile according a gradient volume. Tizoxanide and topiramate (internal standard) were monitored utilizing negative electron spray ionization in multiple reaction monitoring mode. The methods were validated to be precise and accurate within the dynamic range of 5-1000 ng/mL and 0.2-50 ng/g for plasma and brain tissue samples, respectively. The lower limit of quantitation of the method was 0.2 ng/g, which was far more sensitive than all existing methods to quantify tizoxanide in biological samples. Application performed on rats exhibited that the brain-to-plasma ratio of tizoxanide ranged from 3.16 to 26.86% in 1 h after administration of 10 mg/kg nitazoxanide., (© 2019 John Wiley & Sons, Ltd.)

In this paper, the dynamic behavior of bovine brain tissue, measured from in-vitro unconfined compression tests, is examined and represented through a viscoelastic biphasic model. The experiments have been carried out under three compression speeds of 10, 100, and 1000 mm/s. The results exhibited significant rate-dependent behavior. The brain tissue is modeled as a biphasic continuum consisting of a compressible solid matrix, fully saturated with an incompressible interstitial fluid. The governing equations based on conservation of mass and momentum are used to describe the solid-fluid interactions. An inverse scheme is employed in which a finite element model runs iteratively to optimize constitutive constants. The obtained material parameters of the proposed biphasic model show relatively good agreement (R 2  ≥ 0.96) with the experimental tissue mechanical responses at different rates. The model can successfully capture the key aspects of the rate-dependency for both solid and fluid phases under large strain deformation. This poro-hyper viscoelastic model can effectively estimate the global and local rate-dependent tissue deformations, the spatial variations in pore spaces, hydrostatic pressure as well as fluid diffusion through the tissue., Competing Interests: Declaration of competing interest The authors declare that there are no conflicts of interest., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

We recently developed the National Dementia Biobank in México (BioBanco Nacional de Demencias, BND) as a unit for diagnosis, research, and tissue transfer for research purposes. BND is associated with the Facultad de Estudios Superiores Cuautitlán, Universidad Nacional Autónoma de Mexico (UNAM), Mexico. The donation of fluids, brain, and other organs of deceased donors is crucial for understanding the underlying mechanisms of neurodegenerative diseases and for the development of successful treatment. Our laboratory research focuses on 1) analysis of the molecular processing of the proteins involved in those neurodegenerative diseases termed tauopathies and 2) the search for biomarkers for the non-invasive and early diagnosis of Alzheimer's disease.

Detailed knowledge about the mechanical properties of brain can improve numerical modeling of the brain under various loading conditions. The success of this modeling depends on constitutive model and reliable extraction of its material constants. The isotropy of the brain tissue is a key factor which affects the form of constitutive models. In this study, compression tests were performed on different parts of the sheep brain tissue. Also, the digital image correlation (DIC) method was utilized to investigate the direction dependency of brain parts considering their microstructures. To this aim, the DIC method was employed to measure the transverse strain of two lateral sides of the tissue samples. The results of DIC method revealed that the brain stem and corona radiata were isotropic, while the mixed white and gray matter showed an unrepeatable behavior depending on the extracted sample. To examine and validate DIC method, stress-strain diagrams were also used to investigate the isotropy. It could be concluded that axonal fibers had no reinforcing role in the brain tissue. Furthermore, the DIC method indicated incompressibility of the brain tissue. Then, the significance of using a correct method to extract the material constants of brain was discussed. In other words, the effect of the real boundary conditions in experiments, which was neglected in most previous studies, was taken into account here. Finally, the particle swarm optimization algorithm along with the finite element modeling was used to estimate the hyper-viscoelastic constants of different parts of the brain tissue.

Background: Optimisation of tissue processing procedures in preclinical studies reduces the number of animals used and allows integrated multilevel study in the same sample. Multiple extraction of different biomolecules from the same sample has several limitations., New Method: Using brain samples from rats subjected to ischemic stroke, we combined lyophilisation of flash-frozen tissue, mechanical pulverisation and cryopreservation in a method to optimise tissue handling and preservation for independent RNA or protein single-extract methods, and subsequent RT-qPCR or Western blot analyses., Results: Lyophilisation resulted in 70% tissue weight loss. RNA (OD 260/280 ∼1.8) and protein yields were similar in non-ischemic and ischemic brain samples, subjected to either flash freezing (FF) or flash freezing followed by lyophilisation (FF + Lyo). RNA transcription of reference genes (Actb and Rn18s), expression of housekeeping proteins (β-actin and α-tubulin), and mRNA overexpression of stroke-regulated genes (Nos2, Mmp9 and Tnfa) was similar in FF and FF + Lyo samples., Comparison With Existing Method(s): Contrary to high heat stress of baking method in a drying oven, lyophilisation maintains the integrity of dried samples for subsequent extractions and analyses. Sample lyophilisation allows different manual representative extractions/analyses from the same rat, it is much cheaper than using commercial kits, and shows higher yields that multiple manual or kit-based extractions., Conclusions: The lyophilisation-based method for different biomolecule single-extractions from tissue powder aliquots, representing the same rat brain sample, is sample saving, contributes to the reduction principle in animal research, and allows coordinated analysis for accurate correlations between the transcriptome and proteome in stroke and other neuroscience research., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Biomechanical cues guide proliferation, growth and maturation of neurons. Yet the molecules that shape the brain's biomechanical properties are unidentified and the relationship between neural development and viscoelasticity of brain tissue remains elusive. Here we combined novel in-vivo tomoelastography and ex-vivo proteomics to investigate whether viscoelasticity of the mouse brain correlates with protein alterations within the critical phase of brain maturation. For the first time, high-resolution atlases of viscoelasticity of the mouse brain were generated, revealing that (i) brain stiffness increased alongside progressive accumulation of microtubular structures, myelination, cytoskeleton linkage and cell-matrix attachment, and that (ii) viscosity-related tissue fluidity decreased alongside downregulated actin crosslinking and axonal organization. Taken together, our results show that brain maturation is associated with a shift of brain mechanical properties towards a more solid-rigid behavior consistent with reduced tissue fluidity. This shift appears to be driven by several molecular processes associated with myelination, cytoskeletal crosslinking and axonal organization. STATEMENT OF SIGNIFICANCE: The viscoelastic properties of brain tissue shape the environment in which neurons proliferate, grow, and mature. In the present study, novel tomoelastography was used to spatially map tissue mechanical properties of the in-vivo mouse brain during maturation. In vivo tomoelastography was also combined with ex vivo mass spectrometry proteomic analysis to identify the molecules which shape the biomechanical properties of brain tissue. With the combined technique, we observed that brain maturation is associated with a shift of brain mechanical properties towards a more solid-rigid behavior consistent with reduced tissue fluidity which is driven by multiple molecular processes. We believe that this shift of brain mechanical properties discovered in our study reflects a fundamental biophysical signature of brain maturation., (Copyright © 2019 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

1. Antiretroviral concentrations in cerebrospinal fluid (CSF) are used as surrogate for brain tissue, although sparse data support this. We quantified antiretrovirals in brain tissue across preclinical models, compared them to CSF, and calculated 90% inhibitory quotients (IQ 90 ) for nonhuman primate (NHP) brain tissue. Spatial distribution of efavirenz was performed by mass-spectrometry imaging (MSI). 2. HIV or RT-SHIV-infected and uninfected animals from two humanized mouse models (hemopoietic-stem cell/RAG2-, n  = 36; bone marrow-liver-thymus/BLT, n  =13) and an NHP model (rhesus macaque, n  =18) were dosed with six antiretrovirals. Brain tissue, CSF (NHPs), and plasma were collected at necropsy. Drug concentrations were measured by LC-MS/MS. Rapid equilibrium dialysis determined protein binding in NHP brain. 3. Brain tissue penetration of most antiretrovirals were >10-fold lower ( p  < 0.02) in humanized mice than NHPs. NHP CSF concentrations were >13-fold lower ( p  <0.02) than brain tissue with poor agreement except for efavirenz ( r  = 0.91, p  = 0.001). Despite 97% brain tissue protein binding, efavirenz achieved IQ 90 >1 in all animals and 2-fold greater white versus gray matter concentration. 4. Brain tissue penetration varied across animal models for all antiretrovirals except raltegravir, and extrapolating brain tissue concentrations between models should be avoided. With the exception of efavirenz, CSF is not a surrogate for brain tissue concentrations.

Existing brain region segmentation algorithms based on deep convolutional neural networks (CNN) are inefficient for object boundary segmentation. In order to enhance the segmentation accuracy of brain tissue, this paper proposed an object region segmentation algorithm that combines pixel-level information and semantic information. Firstly, we extract semantic information by CNN with the attention module and get the coarse segmentation results through a specific pixel-level classifier. Then, we exploit conditional random fields to model the relationship between the underlying pixels so as to get local features. Finally, the semantic information and the local pixel-level information are respectively used as the unary potential and the binary potential of the Gibbs distribution, and the combination of both can obtain the fine region segmentation algorithm based on the fusion of pixel-level information and the semantic information. A large number of qualitative and quantitative test results show that our proposed algorithm has higher precision than the existing state-of-the-art deep feature models, which can better solve the problem of rough edge segmentation and produce good 3D visualization effect.

In both finite element and physical surrogate models of head blast injury, accurate material properties of the brain and/or tissue simulants are necessary to ensure biofidelity in predicted response. Thus, there is a need for experimental comparisons between tissue and simulant materials under the same experimental conditions. This study compares the response of porcine brain tissue and a variety of brain tissue simulants in quasi-static and sinusoidal compression tests. Fresh porcine brain tissue was obtained from a local abattoir and tested within 4 h post mortem. Additionally, the effect of post mortem time was investigated by comparing samples stored at room temperature and stored frozen (-18 °C), at various time intervals. The brain tissue simulants tested were bovine gelatin (3%, 5%, and 10% concentration), agarose gelatin (e0.4%, 0.6%, 0.8% concentration), and Sylgard 527. The experiments were performed using a DMA apparatus (TA Instruments Q800). The quasi-static compression data were fit to Ogden hyperelastic functions so that parameters could be compared. It was found that bovine gelatin at 3% and 5% concentration demonstrated the closest response to brain tissue in quasi-static compression. Conversely, in sinusoidal compression, the agarose gel and Sylgard 527 were found to be in closer agreement with the tissue, than bovine gel. In terms of post mortem time and storage, there was no statistically significant difference detected in the response of tissue samples after 48 h, regardless of storage method. However, samples stored at room temperature after 48 h appeared to demonstrate a reduction in stiffness., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Ultra-high-resolution imaging mass spectrometry using matrix-assisted laser desorption ionization (MALDI) MS coupled to a Fourier transform ion cyclotron resonance (FTICR) mass analyzer is a powerful technique for the visualization of small molecule distribution within biological tissues. The FTICR MS provides ultra-high resolving power and mass accuracy that allows large molecular coverage and molecular formula assignments, both essential for untargeted metabolomics analysis. These performances require fine optimizations of the MALDI FTICR parameters. In this context, this study proposes a new strategy, using experimental design, for the optimization of ion transmission voltages and MALDI parameters, for tissue untargeted metabolomics analysis, in both positive and negative ionization modes. These experiments were conducted by assessing the effects of nine factors for ion transmission voltages and four factors for MALDI on the number of peaks, the weighted resolution, and the mean error within m/z 150-1000 mass range. For this purpose, fractional factorial designs were used with multiple linear regression (MLR) to evaluate factor effects and to optimize parameter values. The optimized values of ion transmission voltages (RF amplitude TOF, RF amplitude octopole, frequency transfer optic, RF frequency octopole, deflector plate, funnel 1, skimmer, funnel RF amplitude, time-of-flight, capillary exit), MALDI parameters (laser fluence, number of laser shots), and detection parameters (data size, number of scans) led to an increase of 32% and 18% of the number of peaks, an increase of 8% and 39% of the resolution, and a decrease of 56% and 34% of the mean error in positive and negative ionization modes, respectively. Graphical abstract.