Human ß-Defensin 3 Inhibition of P. gingivalis LPS-Induced IL-1ß Production by BV-2 Microglia through Suppression of Cathepsins B and L.

Cathepsin B (CatB) is thought to be essential for the induction of Porphyromonas gingivalis lipopolysaccharide (Pg LPS)-induced Alzheimer's disease-like pathologies in mice, including interleukin-1ß (IL-1ß) production and cognitive decline. However, little is known about the role of CatB in Pg virulence factor-induced IL-1ß production by microglia. We first subjected IL-1ß-luciferase reporter BV-2 microglia to inhibitors of Toll-like receptors (TLRs), IκB kinase, and the NLRP3 inflammasome following stimulation with Pg LPS and outer membrane vesicles (OMVs). To clarify the involvement of CatB, we used several known CatB inhibitors, including CA-074Me, ZRLR, and human ß-defensin 3 (hBD3). IL-1ß production in BV-2 microglia induced by Pg LPS and OMVs was significantly inhibited by the TLR2 inhibitor C29 and the IκB kinase inhibitor wedelolactonne, but not by the NLRPs inhibitor MCC950. Both hBD3 and CA-074Me significantly inhibited Pg LPS-induced IL-1ß production in BV-2 microglia. Although CA-074Me also suppressed OMV-induced IL-1ß production, hBD3 did not inhibit it. Furthermore, both hBD3 and CA-074Me significantly blocked Pg LPS-induced nuclear NF-κB p65 translocation and IκBα degradation. In contrast, hBD3 and CA-074Me did not block OMV-induced nuclear NF-κB p65 translocation or IκBα degradation. Furthermore, neither ZRLR, a specific CatB inhibitor, nor shRNA-mediated knockdown of CatB expression had any effect on Pg virulence factor-induced IL-1ß production. Interestingly, phagocytosis of OMVs by BV-2 microglia induced IL-1ß production. Finally, the structural models generated by AlphaFold indicated that hBD3 can bind to the substrate-binding pocket of CatB, and possibly CatL as well. These results suggest that Pg LPS induces CatB/CatL-dependent synthesis and processing of pro-IL-1ß without activation of the NLRP3 inflammasome. In contrast, OMVs promote the synthesis and processing of pro-IL-1ß through CatB/CatL-independent phagocytic mechanisms. Thus, hBD3 can improve the IL-1ß-associated vicious inflammatory cycle induced by microglia through inhibition of CatB/CatL.

Drug discovery and development for fibromyalgia using practical biomarkers throughout the process from relevant animal models to patients.

INTRODUCTION: Fibromyalgia (FM) is a chronic pain condition characterized by widespread pain and complex comorbidities with a high unmet medical need. Given few past successes in the launch of analgesics with new mechanisms, the implementation of practical biomarkers for drug discovery and development would be necessary to rationally create innovative drugs for chronic pain conditions, including FM. AREAS COVERED: This review surveys the evidence on pathophysiology of FM and the findings regarding the pathophysiology-associated practical biomarker candidates in body fluids (e.g. blood) from the studies in FM patients. This review also summarizes the most commonly used animal models simulating key aspects of clinical FM features. Finally, a strategy for rationally creating innovative drugs for FM is discussed. EXPERT OPINION: Drug discovery and development for FM targeting immune dysregulation/inflammation would be a viable strategy based on the availability of the pathophysiology-associated practical biomarkers (e.g. serum interleukins), which monitor the efficacy of interventions and/or identify responders based on the matching pathophysiology throughout the process from animal models to patients. This strategy could lead to a breakthrough in the development of drugs for FM, a chronic pain condition.

[Transition of Standard Dosimetry of Absorbed Dose to Water in External Beam Radiotherapy [Part 1]].

The radiotherapy is performed with the aim of delivering the optimal dose to the target volume with minimal side effect of surrounding normal tissue. For this purpose, quality assurance is essential to ensure that the target volume is correctly irradiated in the optimal geometrical arrangement, and the absorbed dose evaluation is essential to ensure that the prescribed dose is correctly delivered. The absorbed doses are generally evaluated using a small cavity ionization chamber that utilizes gas ionization. For the evaluation of absorbed dose to water using ionization chambers, the national dose and charge standards, ionization chambers and electrometer calibration systems are required. And it is also required standard dosimetry protocol that recommend conditions such as fields, depths, and optimal ionization chambers for the measurement, as well as reliable physical data. This manuscript reviews the transition of standard dosimetry of absorbed dose to water in external beam radiotherapy, including the background of dose standards, ionization chamber calibration systems, units, and physical constants.

THz Gas Sensing Using Terahertz Time-Domain Spectroscopy with Ceramic Architecture.

Ceramic architectures based on chemical vapor deposition (CVD) are used to create unique crystal structures, morphologies, and properties. This study proposed room-temperature THz gas sensing using terahertz time-domain spectroscopy (THz-TDS) with ceramic architectures. We synthesized ceramic films on porous glass. Zinc oxide films were created using atmospheric CVD and amorphous carbon nitride films using the dissociative excitation reaction of BrCN with metastable Ar atoms. The transmission method was used in THz-TDS. A stainless hand-made gas cell with a Si window was applied for THz gas sensing. We defined "phase delay" equals VOC sensing response amount of sensing materials at each duration. Ppm-order THz gas sensing was performed.

Selective Involvement of a Subset of Spinal Dorsal Horn Neurons Operated by a Prodynorphin Promoter in Aß Fiber-Mediated Neuropathic Allodynia-Like Behavioral Responses in Rats.

Mechanical allodynia (pain produced by innocuous stimuli such as touch) is the main symptom of neuropathic pain. Its underlying mechanism remains to be elucidated, but peripheral nerve injury (PNI)-induced malfunction of neuronal circuits in the central nervous system, including the spinal dorsal horn (SDH), is thought to be involved in touch-pain conversion. Here, we found that intra-SDH injection of adeno-associated viral vectors including a prodynorphin promoter (AAV-PdynP) captured a subset of neurons that were mainly located in the superficial laminae, including lamina I, and exhibited mostly inhibitory characteristics. Using transgenic rats that enable optogenetic stimulation of touch-sensing Aß fibers, we found that the light-evoked paw withdrawal behavior and aversive responses after PNI were attenuated by selective ablation of AAV-PdynP-captured SDH neurons. Notably, the ablation had no effect on withdrawal behavior from von Frey filaments. Furthermore, Aß fiber stimulation did not excite AAV-PdynP+ SDH neurons under normal conditions, but after PNI, this induced excitation, possibly due to enhanced Aß fiber-evoked excitatory synaptic inputs and elevated resting membrane potentials of these neurons. Moreover, the chemogenetic silencing of AAV-PdynP+ neurons of PNI rats attenuated the Aß fiber-evoked paw withdrawal behavior and c-FOS expression in superficial SDH neurons. Our findings suggest that PNI renders AAV-PdynP-captured neurons excitable to Aß fiber stimulation, which selectively contributes to the conversion of Aß fiber-mediated touch signal to nociceptive. Thus, reducing the excitability of AAV-PdynP-captured neurons may be a new option for the treatment of neuropathic allodynia.

Chemogenetic silencing of spinal cord-projecting cortical neurons attenuates Aß fiber-derived neuropathic allodynia in mice.

Mechanical allodynia (pain caused by innocuous mechanical stimulation) is a hallmark symptom of neuropathic pain occurring following peripheral nerve injury (PNI). Using a transgenic mouse line, in which myelinated primary afferents, including Aß fibers, express channelrhodopsin-2, we found that illumination of the plantar skin of mice following PNI produced an Aß fiber-mediated pain-like withdrawal behavior and increased c-FOS+ neurons in the superficial spinal dorsal horn (SDH). These two responses were attenuated by chemogenetic silencing of primary sensory cortex (S1) neurons projecting directly to the SDH. These findings indicate that spinally projecting cortical S1 neurons contribute to Aß fiber-derived neuropathic allodynia.

The Role of Microglial Purinergic Receptors in Pain Signaling.

Pain is an essential modality of sensation in the body. Purinergic signaling plays an important role in nociceptive pain transmission, under both physiological and pathophysiological conditions, and is important for communication between both neuronal and non-neuronal cells. Microglia and astrocytes express a variety of purinergic effectors, and a variety of receptors play critical roles in the pathogenesis of neuropathic pain. In this review, we discuss our current knowledge of purinergic signaling and of the compounds that modulate purinergic transmission, with the aim of highlighting the importance of purinergic pathways as targets for the treatment of persistent pain.

Efficacy of tangential irradiation with volumetric modulated arc therapy on scalp angiosarcoma using medical linac.

PURPOSE: To increase the superficial dose and reduce the brain dose for radiotherapy of scalp angiosarcoma, we propose a novel irradiation technique of tangential irradiation volumetric modulated arc therapy (TI-VMAT). METHODS: TI-VMAT and the conventional VMAT treatment plans for thirteen scalp angiosarcoma patients were created with a prescribed dose of 70 Gy. Each treatment was normalized to cover 95% of the planning target volume (PTV) with its prescribed dose. To realize TI-VMAT, an avoidance structure (AS) function was applied. AS was defined as a contour subtracted PTV by a certain space from the brain contour. TI-VMAT treatment plans for six different spaces between PTV and AS were developed and compared with the conventional VMAT treatment plan with respect to the following dosimetric parameters: homogeneity index (HI) and conformity index (CI) of the PTV, mean brain dose, and brain volume irradiated with 20% (V20% [cc]), 40% (V40% [cc]), 60% (V60% [cc]), 80% (V80% [cc]), and 100% (V100% [cc]) of the prescribed dose. RESULTS: HI and CI were comparable between TI-VMAT and the conventional VMAT, the mean brain dose for TI-VMAT with AS defined by a space of 2.0 cm and jaw tracking was 14.27 Gy, which was significantly lower than that for the conventional VMAT (21.20 Gy). In addition, dosimetric parameters such as V20% [cc] were significantly suppressed compared to those for high doses. CONCLUSION: Our proposed irradiation technique TI-VMAT shows the potential to reduce radiation doses in the brain with maintaining higher dose coverage on the PTV.

Evaluation of auto-segmentation accuracy of cloud-based artificial intelligence and atlas-based models.

BACKGROUND: Contour delineation, a crucial process in radiation oncology, is time-consuming and inaccurate due to inter-observer variation has been a critical issue in this process. An atlas-based automatic segmentation was developed to improve the delineation efficiency and reduce inter-observer variation. Additionally, automated segmentation using artificial intelligence (AI) has recently become available. In this study, auto-segmentations by atlas- and AI-based models for Organs at Risk (OAR) in patients with prostate and head and neck cancer were performed and delineation accuracies were evaluated. METHODS: Twenty-one patients with prostate cancer and 30 patients with head and neck cancer were evaluated. MIM Maestro was used to apply the atlas-based segmentation. MIM Contour ProtégéAI was used to apply the AI-based segmentation. Three similarity indices, the Dice similarity coefficient (DSC), Hausdorff distance (HD), and mean distance to agreement (MDA), were evaluated and compared with manual delineations. In addition, radiation oncologists visually evaluated the delineation accuracies. RESULTS: Among patients with prostate cancer, the AI-based model demonstrated higher accuracy than the atlas-based on DSC, HD, and MDA for the bladder and rectum. Upon visual evaluation, some errors were observed in the atlas-based delineations when the boundary between the small bowel or the seminal vesicle and the bladder was unclear. For patients with head and neck cancer, no significant differences were observed between the two models for almost all OARs, except small delineations such as the optic chiasm and optic nerve. The DSC tended to be lower when the HD and the MDA were smaller in small volume delineations. CONCLUSIONS: In terms of efficiency, the processing time for head and neck cancers was much shorter than manual delineation. While quantitative evaluation with AI-based segmentation was significantly more accurate than atlas-based for prostate cancer, there was no significant difference for head and neck cancer. According to the results of visual evaluation, less necessity of manual correction in AI-based segmentation indicates that the segmentation efficiency of AI-based model is higher than that of atlas-based model. The effectiveness of the AI-based model can be expected to improve the segmentation efficiency and to significantly shorten the delineation time.

Direct energy spectrum measurement of X-ray from a clinical linac.

A realistic X-ray energy spectrum is essential for accurate dose calculation using the Monte Carlo (MC) algorithm. An energy spectrum for dose calculation in the radiation treatment planning system is modeled using the MC algorithm and adjusted to obtain acceptable agreement with the measured percent depth dose (PDD) and off-axis ratio. The simulated energy spectrum may not consistently reproduce a realistic energy spectrum. Therefore, direct measurement of the X-ray energy spectrum from a linac is necessary to obtain a realistic spectrum. Previous studies have measured low photon fluence directly, but the measurement was performed with a nonclinical linac with a thick target and a long target-to-detector distance. In this study, an X-ray energy spectrum from a clinical linac was directly measured using a NaI(Tl) scintillator at an ultralow dose rate achieved by adjusting the gun grid voltage. The measured energy spectrum was unfolded by the Gold algorithm and compared with a simulated spectrum using statistical tests. Furthermore, the PDD was calculated using an unfolded energy spectrum and a simulated energy spectrum was compared with the measured PDD to evaluate the validity of the unfolded energy spectrum. Consequently, there was no significant difference between the unfolded and simulated energy spectra by nonparametric, Wilcoxon's rank-sum, chi-square, and two-sample Kolmogorov-Smirnov tests with a significance level of 0.05. However, the PDD calculated from the unfolded energy spectrum better agreed with the measured compared to the calculated PDD results from the simulated energy spectrum. The adjustment of the incident electron parameters using MC simulation is sensitive and takes time. Therefore, it is desirable to obtain the energy spectrum by direct measurement. Thus, a method to obtain the realistic energy spectrum by direct measurement was proposed in this study.

In Situ Chemical Lithiation Transforms Diamond-Like Carbon into an Ultrastrong Ion Conductor for Dendrite-Free Lithium-Metal Anodes.

Lithium (Li)-metal anodes are of great promise for next-generation batteries due to their high theoretical capacity and low redox potential. However, Li-dendrite growth during cycling imposes a tremendous safety concern on the practical application of Li-metal anodes. Herein, an effective approach to suppress Li-dendrite growth by coating a polypropylene (PP) separator with a thin layer of ultrastrong diamond-like carbon (DLC) is reported. Theoretical calculations indicate that the DLC coating layer undergoes in situ chemical lithiation once assembled with the lithium-metal anode, transforming the DLC/PP separator into an excellent 3D Li-ion conductor. This in situ lithiated DLC/PP separator can not only mechanically suppress Li-dendrite growth by its intrinsically high modulus (≈100 GPa), but also uniformly redistributes Li ions to render dendrite-free lithium deposition. The twofold effects of the DLC/PP separator result in stable cycling of lithium plating/stripping (over 4500 h) at a high current density of 3 mA cm-2 . Remarkably, this approach enables more than 1000 stable cycles at 5 C with a capacity retention of ≈71% in a Li || LiFePO4 coin cell and more than 200 stable cycles at 0.2 C in a Li || LiNi0.5 Co0.3 Mn0.2 O2 pouch cell with cathode mass loading of ≈9 mg cm-2 .

New Inhibitory Effects of Cilnidipine on Microglial P2X7 Receptors and IL-1ß Release: An Involvement in its Alleviating Effect on Neuropathic Pain.

P2X7 receptors (P2X7Rs) belong to a family of ATP-gated non-selective cation channels. Microglia represent a major cell type expressing P2X7Rs. The activation of microglial P2X7Rs causes the release of pro-inflammatory cytokines such as interleukin-1ß (IL-1ß). This response has been implicated in neuroinflammatory states in the central nervous system and in various diseases, including neuropathic pain. Thus, P2X7R may represent a potential therapeutic target. In the present study, we screened a chemical library of clinically approved drugs (1979 compounds) by high-throughput screening and showed that the Ca2+ channel blocker cilnidipine has an inhibitory effect on rodent and human P2X7R. In primary cultured rat microglial cells, cilnidipine inhibited P2X7R-mediated Ca2+ responses and IL-1ß release. Moreover, in a rat model of neuropathic pain, the intrathecal administration of cilnidipine produced a reversal of nerve injury-induced mechanical hypersensitivity, a cardinal symptom of neuropathic pain. These results point to a new inhibitory effect of cilnidipine on microglial P2X7R-mediated inflammatory responses and neuropathic pain, proposing its therapeutic potential.

A subset of spinal dorsal horn interneurons crucial for gating touch-evoked pain-like behavior.

A cardinal, intractable symptom of neuropathic pain is mechanical allodynia, pain caused by innocuous stimuli via low-threshold mechanoreceptors such as Aß fibers. However, the mechanism by which Aß fiber-derived signals are converted to pain remains incompletely understood. Here we identify a subset of inhibitory interneurons in the spinal dorsal horn (SDH) operated by adeno-associated viral vectors incorporating a neuropeptide Y promoter (AAV-NpyP+) and show that specific ablation or silencing of AAV-NpyP+ SDH interneurons converted touch-sensing Aß fiber-derived signals to morphine-resistant pain-like behavioral responses. AAV-NpyP+ neurons received excitatory inputs from Aß fibers and transmitted inhibitory GABA signals to lamina I neurons projecting to the brain. In a model of neuropathic pain developed by peripheral nerve injury, AAV-NpyP+ neurons exhibited deeper resting membrane potentials, and their excitation by Aß fibers was impaired. Conversely, chemogenetic activation of AAV-NpyP+ neurons in nerve-injured rats reversed Aß fiber-derived neuropathic pain-like behavior that was shown to be morphine-resistant and reduced pathological neuronal activation of superficial SDH including lamina I. These findings suggest that identified inhibitory SDH interneurons that act as a critical brake on conversion of touch-sensing Aß fiber signals into pain-like behavioral responses. Thus, enhancing activity of these neurons may offer a novel strategy for treating neuropathic allodynia.

Properties and Classification of Diamond-Like Carbon Films.

Diamond-like carbon (DLC) films have been extensively applied in industries owing to their excellent characteristics such as high hardness. In particular, there is a growing demand for their use as protective films for mechanical parts owing to their excellent wear resistance and low friction coefficient. DLC films have been deposited by various methods and many deviate from the DLC regions present in the ternary diagrams proposed for sp3 covalent carbon, sp2 covalent carbon, and hydrogen. Consequently, redefining the DLC region on ternary diagrams using DLC coatings for mechanical and electrical components is urgently required. Therefore, we investigate the sp3 ratio, hydrogen content, and other properties of 74 types of amorphous carbon films and present the classification of amorphous carbon films, including DLC. We measured the sp3 ratios and hydrogen content using near-edge X-ray absorption fine structure and Rutherford backscattering-elastic recoil detection analysis under unified conditions. Amorphous carbon films were widely found with nonuniform distribution. The number of carbon atoms in the sp3 covalent carbon without bonding with hydrogen and the logarithm of the hydrogen content were inversely proportional. Further, we elucidated the DLC regions on the ternary diagram, classified the amorphous carbon films, and summarized the characteristics and applications of each type of DLC.

Spinal astrocytes in superficial laminae gate brainstem descending control of mechanosensory hypersensitivity.

Astrocytes are critical regulators of CNS function and are proposed to be heterogeneous in the developing brain and spinal cord. Here we identify a population of astrocytes located in the superficial laminae of the spinal dorsal horn (SDH) in adults that is genetically defined by Hes5. In vivo imaging revealed that noxious stimulation by intraplantar capsaicin injection activated Hes5+ SDH astrocytes via α1A-adrenoceptors (α1A-ARs) through descending noradrenergic signaling from the locus coeruleus. Intrathecal norepinephrine induced mechanical pain hypersensitivity via α1A-ARs in Hes5+ astrocytes, and chemogenetic stimulation of Hes5+ SDH astrocytes was sufficient to produce the hypersensitivity. Furthermore, capsaicin-induced mechanical hypersensitivity was prevented by the inhibition of descending locus coeruleus-noradrenergic signaling onto Hes5+ astrocytes. Moreover, in a model of chronic pain, α1A-ARs in Hes5+ astrocytes were critical regulators for determining an analgesic effect of duloxetine. Our findings identify a superficial SDH-selective astrocyte population that gates descending noradrenergic control of mechanosensory behavior.

Intrinsic braking role of descending locus coeruleus noradrenergic neurons in acute and chronic itch in mice.

Itch is defined as an unpleasant sensation that provokes a desire to scratch. Our understanding of neuronal circuits for itch information transmission and processing in the spinal dorsal horn (SDH) has progressively advanced following the identification of SDH neuron subsets that are crucial for scratching behavior in models of itch. However, little is known about the control of acute and chronic itch by descending signals from the brain to the SDH. In this study, using genetic approaches that enable cell-type and circuit-specific functional manipulation, we reveal an intrinsic potential of locus coeruleus (LC)-noradrenergic (NAergic) neurons that project to the SDH to control acute and chronic itch. Activation and silencing of SDH-projecting LC-NAergic neurons reduced and enhanced scratching behavior, respectively, in models of histamine-dependent and -independent acute itch. Furthermore, in a model of chronic itch associated with contact dermatitis, repetitive scratching behavior was suppressed by the activation of the descending LC-NAergic pathway and by knocking out NA transporters specific to descending LC-NAergic neurons using a CRISPR-Cas9 system. Moreover, patch-clamp recording using spinal slices showed that noradrenaline facilitated inhibitory synaptic inputs onto gastrin-releasing peptide receptor-expressing SDH neurons, a neuronal subset known to be essential for itch transmission. Our findings suggest that descending LC-NAergic signaling intrinsically controls acute and chronic itch and provide potential therapeutic strategies for the treatment of acute and chronic itch.

Estimation of the cable effect in megavoltage photon beam by measurement and Monte Carlo simulation.

PURPOSE: Ionization chambers are widely used for dosimetry with megavoltage photon beams. Several properties of ionization chambers, including the cable effect, polarity effect, and ion recombination loss, are described in standard dosimetry protocols. The cable effect is categorized as the leakage current and Compton current, and careful consideration of these factors has been described not only in reference dosimetry but also in large fields. However, the mechanism of Compton current in the cable has not been investigated thoroughly. The cable effect of ionization chambers in 6 MV X-ray beam was evaluated by measurement, and the mechanism of Compton current was investigated by Monte Carlo simulation. MATERIALS AND METHODS: Four PTW ionization chambers (TM30013, TM31010, TM31014, and TM31016) with the same type of mounted cable, but different ionization volumes, were used to measure output factor (OPF) and cable effect measurement. The OPF was measured to observe any variation resulting from the cable effect. The cable effect was evaluated separately for the leakage current and Compton current, and its charge per absorbed dose to water per cable length was estimated by a newly proposed method. The behavior of electrons and positrons in the core wire was analyzed and the Compton current for the photon beam was estimated by Monte Carlo simulation. RESULTS: In OPF measurement, the difference in the electrometer readings by polarity became obvious for the mini- or microchamber and its difference tended to be larger for a chamber with a smaller ionization volume. For the cable effect measurement, it was determined that the contribution of the leakage current to the cable effect was ignorable, while the Compton current was dominant. The charge due to the Compton current per absorbed dose to water per cable length was estimated to be 0.36 ± 0.03 pC Gy-1  cm-1 for PTW ionization chambers. As a result, the contribution of the Compton current to the electrometer readings was estimated to be 0.002% cm-1 for the Farmer-type, 0.011% cm-1 for the scanning, and 0.088% cm-1 for microchambers, respectively. By the simulation, it was determined that the Compton current for MV x-ray could be explained by not only recoil electrons due to Compton scattering but also positron due to pair production. The Compton current estimated by the difference in outflowing and inflowing charge was 0.45 pC Gy-1  cm-1 and was comparable with the measured value. CONCLUSION: The cable effect, which includes the leakage current and Compton current, was quantitatively estimated for several chambers from measurements, and the mechanism of Compton current was investigated by Monte Carlo simulation. It was determined that the Compton current is a dominant component of the cable effect and its charge is consistently positive and nearly the same, irrespective of the ionization chamber volume. The contribution of Compton current to the electrometer readings was estimated for chambers. The mechanism of Compton current was analyzed and it was confirmed that the Compton current can be estimated from the difference in outflowing and inflowing charge to and from the core wire.

Modulation of P2Y6R expression exacerbates pressure overload-induced cardiac remodeling in mice.

Cardiac tissue remodeling caused by hemodynamic overload is a major clinical outcome of heart failure. Uridine-responsive purinergic P2Y6 receptor (P2Y6R) contributes to the progression of cardiovascular remodeling in rodents, but it is not known whether inhibition of P2Y6R prevents or promotes heart failure. We demonstrate that inhibition of P2Y6R promotes pressure overload-induced sudden death and heart failure in mice. In neonatal cardiomyocytes, knockdown of P2Y6R significantly attenuated hypertrophic growth and cell death caused by hypotonic stimulation, indicating the involvement of P2Y6R in mechanical stress-induced myocardial dysfunction. Unexpectedly, compared with wild-type mice, deletion of P2Y6R promoted pressure overload-induced sudden death, as well as cardiac remodeling and dysfunction. Mice with cardiomyocyte-specific overexpression of P2Y6R also exhibited cardiac dysfunction and severe fibrosis. In contrast, P2Y6R deletion had little impact on oxidative stress-mediated cardiac dysfunction induced by doxorubicin treatment. These findings provide overwhelming evidence that systemic inhibition of P2Y6R exacerbates pressure overload-induced heart failure in mice, although P2Y6R in cardiomyocytes contributes to the progression of cardiac fibrosis.

Mechanical pain of the lower extremity after compression of the upper spinal cord involves signal transducer and activator of transcription 3-dependent reactive astrocytes and interleukin-6.

Chronic pain is one of the main symptoms of spinal disorders such as spinal canal stenosis. A major cause of this pain is related to compression of the spinal cord, and chronic pain can develop at the level of the compressed spinal segment. However, in many patients chronic pain arises in an area that does not correspond to the compressed segment, and the underlying mechanism involved remains unknown. This was investigated in the present study using a mouse model of spinal cord compression in which mechanical pain of the hindpaws develops after compression of the first lumbar segment (L1) of the spinal cord. Compression induced the activation of astrocytes in the L1 spinal dorsal horn (SDH)-but not the L4 SDH that corresponds to the hindpaws-and activated signal transducer and activator of transcription 3 (STAT3). Suppressing reactive astrocytes by expressing a dominant negative form of STAT3 (dnSTAT3) in the compressed SDH prevented mechanical pain. Expression of interleukin (IL)-6 was also upregulated in the compressed SDH, and it was inhibited by astrocytic expression of dnSTAT3. Intrathecal administration of a neutralizing anti-IL-6 antibody reversed the compression-induced mechanical pain. These results suggest that astrocytic STAT3 and IL-6 in the compressed SDH are involved in remote mechanical pain observed in the lower extremity, and may provide a target for treating chronic pain associated with spinal cord compression such as spinal canal stenosis.