Search

To identify genes involved in cerebral infarction, we have employed a forward genetic approach in inbred mouse strains, using quantitative trait loci (QTL) mapping for cerebral infarct volume after middle cerebral artery occlusion. We had previously observed that infarct volume is inversely correlated with cerebral collateral vessel density in most strains. In this study, we expanded the pool of allelic variation among classical inbred mouse strains by utilizing the eight founder strains of the Collaborative Cross and found a wild-derived strain, WSB/EiJ, that breaks this general rule that collateral vessel density inversely correlates with infarct volume. WSB/ EiJ and another wild-derived strain, CAST/EiJ, show the highest collateral vessel densities of any inbred strain, but infarct volume of WSB/EiJ mice is 8.7-fold larger than that of CAST/EiJ mice. QTL mapping between these strains identified four new neuroprotective loci modulating cerebral infarct volume while not affecting collateral vessel phenotypes. To identify causative variants in genes, we surveyed nonsynonymous coding SNPs between CAST/EiJ and WSB/EiJ and found 96 genes harboring coding SNPs predicted to be damaging and mapping within one of the four intervals. In addition, we performed RNA-sequencing for brain tissue of CAST/EiJ and WSB/EiJ mice and identified 79 candidate genes mapping in one of the four intervals showing strain-specific differences in expression. The identification of the genes underlying these neuroprotective loci will provide new understanding of genetic risk factors of ischemic stroke, which may provide novel targets for future therapeutic intervention of human ischemic stroke. [ABSTRACT FROM AUTHOR]

Objective: This experiment aimed to investigate the effect of different sources and inclusion levels of dietary fat on productive performance and egg quality in laying hens raised under hot environmental conditions. Methods: A total of 480 Hy-Line Brown laying hens at 31 wk of age were randomly allotted to 1 of 5 experimental diets. The control diet contained 2,800 kcal/kg nitrogen-corrected apparent metabolizable energy with no fat addition. Four additional diets were prepared by adding 2.0% or 4.0% of animal fat (AF) or soybean oil (SO). Energy and nutrient concentrations were consistent among all diets. Diets were fed to hens for 4 weeks. Average daily room temperature and humidity were 26.7°C±1.52°C and 77.4%±4.50%. The heat stress index was approximately 76, indicating that hens were raised under heat stress conditions. Results: Final body weight (BW) was greater (p<0.05) for hens fed diets containing 2.0% or 4.0% AF than for those fed the control diet or diets containing 2.0% or 4.0% SO. The BW gain and feed intake were greater (p<0.05) for hens fed diets containing additional AF or SO than those fed the control diet. Eggshell thickness was the greatest (p<0.05) for hens fed the control diet, but the least (p<0.05) for hens fed diets containing 4.0% SO. Egg yolk color was the greatest (p<0.05) for hens fed the control diet, but the least (p<0.05) for hens fed diets containing 4.0% SO. Conclusion: Inclusion of supplemental fat (AF and SO) in diets exhibits preventative effects on BW loss for hens raised under hot environmental conditions when energy and nutrient concentrations in diets were maintained. The effects were greater for AF than for SO. However, inclusion of supplemental fat in diets decreases eggshell thickness and egg yolk yellowness, possibly due to a reduction in Ca absorption and intake of egg yolk colorants. [ABSTRACT FROM AUTHOR]

Itching is a common clinical symptom of skin disease that significantly affects a patient's quality of life. Transient receptor potential vanilloid 1 (TRPV1) receptors of keratinocytes and peripheral nerve fibers in skin are involved in the regulation of itching as well as pain. In this study, we investigated whether curcumin, which acts on TRPV1 receptors, affects histamine-induced itching in mice, using behavioral tests and electrophysiological approaches. We found that histamine-induced itching was blocked by topical application of curcumin in a concentration-dependent manner. In ex-vivo recordings, histamine-induced discharges of peripheral nerves were reduced by the application of curcumin, indicating that curcumin acts directly on peripheral nerves. Additionally, curcumin blocked the histamine-induced inward current via activation of TRPV1 (curcumin IC50=523 nM). However, it did not alter chloroquine-induced itching behavior in mice, which is associated with transient receptor potential ankyrin 1 (TRPA1). Taken together, our results suggest that histamine-induced itching can be blocked by topical application of curcumin through the inhibitory action of curcumin on TRPV1 receptors in peripheral nerves. [ABSTRACT FROM AUTHOR]

Mammalian target of rapamycin complex 1 (mTORC1), a nutrient sensor and central controller of cell growth and proliferation, is altered in various models of Alzheimer's disease (AD). Even less studied or understood in AD is mammalian target of rapamycin complex 2 (mTORC2) that influences cellular metabolism, in part through the regulations of Akt/PKB and SGK. Dysregulation of insulin/PI3K/Akt signaling is another important feature of AD pathogenesis. We found that both total mTORC1 and C2 protein levels and individual C1 and C2 enzymatic activities were decreased in human AD brain samples. In two rodent AD models, mTORC1 and C2 activities were also decreased. In a neuronal culture model of AD characterized by accumulation of cellular amyloid-β (Aβ)42, mTORC1 activity was reduced. Autophagic vesicles and markers were correspondingly increased and new protein synthesis was inhibited, consistent with mTORC1 hypofunction. Interestingly, mTORC2 activity in neural culture seemed resistant to the effects of intracellular amyloid. In various cell lines, Aβ expression provoked insulin resistance, characterized by inhibition of stimulated Akt phosphorylation, and an increase in negative mTORC1 regular, p-AMPK, itself a nutrient sensor. Rapamycin decreased phospho-mTOR and to lesser degree p-Rictor. This further suppression of mTORC1 activity protected cells from Aβ-induced toxicity and insulin resistance. More striking, Rictor over-expression fully reversed the Aβ-effects on primary neuronal cultures. Finally, using in vitro assay, Rictor protein addition completely overcame oligomeric Aβ-induced inhibition of the PDK-Akt activation step. We conclude that striking a new balance by restoring mTORC2 abundance and/or inhibition of mTORC1 has therapeutic potential in AD. [ABSTRACT FROM AUTHOR]

Manipulation of magnetization by electric field is a central goal of spintronics because it enables energy-efficient operation of spin-based devices. Spin wave devices are promising candidates for low-power information processing, but a method for energy-efficient excitation of short-wavelength spin waves has been lacking. Here we show that spin waves in nanoscale magnetic tunnel junctions can be generated via parametric resonance induced by electric field. Parametric excitation of magnetization is a versatile method of short-wavelength spin wave generation, and thus, our results pave the way toward energy-efficient nanomagnonic devices. [ABSTRACT FROM AUTHOR]

The analgesic mechanism of opioids is known to decrease the excitability of substantia gelatinosa (SG) neurons receiving the synaptic inputs from primary nociceptive afferent fiber by increasing inwardly rectifying K+ current. In this study, we examined whether a µ-opioid agonist, [D-Ala2,N-Me-Phe4, Gly5-ol]-enkephalin (DAMGO), affects the two-pore domain K+ channel (K2P) current in rat SG neurons using a slice whole-cell patch clamp technique. Also we confirmed which subtypes of K2P channels were associated with DAMGO-induced currents, measuring the expression of K2P channel in whole spinal cord and SG region. DAMGO caused a robust hyperpolarization and outward current in the SG neurons, which developed almost instantaneously and did not show any time-dependent inactivation. Half of the SG neurons exhibited a linear I~V relationship of the DAMGO-induced current, whereas rest of the neurons displayed inward rectification. In SG neurons with a linear I~V relationship of DAMGO-induced current, the reversal potential was close to the K+ equilibrium potentials. The mRNA expression of TWIK (tandem of pore domains in a weak inwardly rectifying K+ channel) related acid-sensitive K+ channel (TASK) 1 and 3 was found in the SG region and a low pH (6.4) significantly blocked the DAMGO-induced K+ current. Taken together, the DAMGO-induced hyperpolarization at resting membrane potential and subsequent decrease in excitability of SG neurons can be carried by the two-pore domain K+ channel (TASK1 and 3) in addition to inwardly rectifying K+ channel. [ABSTRACT FROM AUTHOR]

Risk for ischemic stroke has a strong genetic basis, but heritable factors also contribute to the extent of damage after a stroke has occurred. We previously identified a locus on distal mouse chromosome 7 that contributes over 50% of the variation in postischemic cerebral infarct volume observed between inbred strains. Here, we used ancestral haplotype analysis to fine-map this locus to 12 candidate genes. The gene encoding the IL-21 receptor (Il21r) showed a marked difference in strain-specific transcription levels and coding variants in neonatal and adult cortical tissue. Collateral vessel connections were moderately reduced in Il21r-deficient mice, and cerebral infarct volume increased 2.3-fold, suggesting that Il21r modulates both collateral vessel anatomy and innate neuroprotection. In brain slice explants, oxygen deprivation (OD) activated apoptotic pathways and increased neuronal cell death in IL-21 receptor-deficient (IL-21R-deficient) mice compared with control animals. We determined that the neuroprotective effects of IL-21R arose from signaling through JAK/STAT pathways and upregulation of caspase 3. Thus, natural genetic variation in murine Il21r influences neuronal cell viability after ischemia by modulating receptor function and downstream signal transduction. The identification of neuroprotective genes based on naturally occurring allelic variations has the potential to inform the development of drug targets for ischemic stroke treatment. [ABSTRACT FROM AUTHOR]

The GluA2 subunit of AMPA-type glutamate receptors (AMPARs) regulates excitatory synaptic transmission in neurons. In addition, the transsynaptic cell adhesion molecule N-cadherin controls excitatory synapse function and stabilizes dendritic spine structures. At postsynaptic membranes, GluA2 physically binds N-cadherin, underlying spine growth and synaptic modulation. We report that N-cadherin binds to PSD-95/SAP90/DLG/ZO-1 (PDZ) domain 2 of the glutamate receptor interacting protein 1 (GRIP1) through its intracellular C terminus. N-cadherin and GluA2-containing AMPARs are presorted to identical transport vesicles for dendrite delivery, and live imaging reveals cotransport of both proteins. The kinesin KIF5 powers GluA2/N-cadherin codelivery by using GRIP1 as a multilink interface. Notably, GluA2 and N-cadherin use different PDZ domains on GRIP1 to simultaneously bind the transport complex, and interference with either binding motif impairs the turnover of both synaptic cargoes. Depolymerization of microtubules, deletion of the KIF5 motor domain, or specific blockade of AMPAR exocytosis affects delivery of GluA2/N-cadherin vesicles. At the functional level, interference with this cotransport reduces the number of spine protrusions and excitatory synapses. Our data suggest the concept that the multi-PDZ-domain adaptor protein GRIP1 can act as a scaffold at trafficking vesicles in the combined delivery of AMPARs and N-cadherin into dendrites. [ABSTRACT FROM AUTHOR]

The effect of the binders, such as silica, alumina, and aluminum phosphate solution (APS), was studied on the acidity and catalytic performance of HZSM-5 zeolite (SiO2/Al2O3 = 80) in methanol-to-propylene (MTP) process. The strong acidity of catalyst increased slightly with alumina binder but decreased with silica and APS binders. It is noted that the catalyst with APS binder showed the highest bulk crush strength. Catalytic performance of the alumina or silica bound catalyst was comparable to that of pure HZSM-5 catalyst while that of APS bound catalyst was completely different depending on the binder content. Low content (10 wt%) of APS resulted in a dramatically enhanced propylene selectivity (>40 C mol%) due to the decrease in strong acidity. On the other hand with high content (>20 wt%) of APS, methanol was mostly dehydrated into dimethylether without further transformation into hydrocarbons even if the mechanical strength was significantly improved. The binary binder system was proposed not only to improve the mechanical strength of catalyst with small amount of APS binder but also to keep a high propylene selectivity. The catalyst bound with APS-and-alumina or APS-and-silica showed a comparable propylene selectivity to that of HZSM-5 with the 10 wt% of APS single binder while demonstrated much higher bulk crush strengths. [ABSTRACT FROM AUTHOR]

Major depressive disorder (MDD) is a devastating disease that arises in a background of environmental risk factors, such as chronic stress, that produce reactive oxygen species (ROS) in the brain. The chronic stress-induced ROS production involves Ca2+ signals; however, the mechanism is poorly understood. Transient receptor potential melastatin type 2 (TRPM2) is a Ca2+-permeable cation channel that is highly expressed in the brain. Here we show that in animal models of chronic unpredictable stress (CUS), deletion of TRPM2 (Trpm2-/-) produces antidepressant-like behaviors in mice. This phenotype correlates with reduced ROS, ROSinduced calpain activation, and enhanced phosphorylation of two Cdk5 targets including synapsin 1 and histone deacetylase 5 that are linked to synaptic function and gene expression, respectively. Moreover, TRPM2 mRNA expression is increased in hippocampal tissue samples from patients with MDD. Our findings suggest that TRPM2 is a key agent in stress-induced depression and a possible target for treating depression. [ABSTRACT FROM AUTHOR]

Abnormal aggregation of α-Synuclein (αS) drives cell loss and produces intracellular inclusions in synucleinopathies, which may be due to insufficient degradation resulting from either a dysfunctional ubiquitin-proteasome system (UPS) or a constant production of mutated αS that overwhelms the UPS. Recent in vitro studies suggest that the autophagylysosome pathway is also involved in αS degradation. On the other hand, synucleinopathies are associated with microglial activation and infiltration. In this postmortem study, we aimed to investigate the relationship between microglial activity and αS disposition in serial patients with cerebral infarct-induced Wallerian degeneration (CI-WD) of the corticospinal fibers. Immunoreactivity for αS was found in activated microglias localized in the CI-WD regions of seven patients (mean 65.6 ± 9.3 years), but not five neuropathologically normal subjects (mean 70.4 ± 8.0 years). No immunoreactivity for ubiquitin or neurofilament protein was present in the microglias. Confocal analysis of double-immunofluorescence staining confirmed the colocalization of αS with the microglial marker. Our results suggest endocytosis (phagocytosis and/or pinocytosis) of αS into activated microglias during the process of CI-WD. This finding indicates that αS may alternatively undergo microglia-associated lysosomal proteolysis for degradation. [ABSTRACT FROM AUTHOR]

TRPV1, a ligand-gated ion channel expressed in nociceptive sensory neurons is modulated by a variety of intracellular signaling pathways. Dopamine is a neurotransmitter that plays important roles in motor control, cognition, and pain modulation in the CNS, and acts via a variety of dopamine receptors (D1R eD5R), a class of GPCRs. Although nociceptive sensory neurons express D1-like receptors, very little is known about the effect of dopamine on TRPV1 in the peripheral nervous system. Therefore, in this study, we examined the effects of D1R activation on TRPV1 in mouse DRG neurons using Ca2+ imaging and immunohistochemical analysis. The D1R agonist SKF-38393 induced reproducible Ca2+ responses via Ca2+ influx through TRPV1 rather than Ca2+ mobilization from intracellular Ca2+ stores. Immunohistochemical analysis revealed co-expression of D1R and TRPV1 in mouse DRG neurons. The PLC-specific inhibitor blocked the SKF-38393-induced Ca2+ response, whereas the PKC, DAG lipase, AC, and PKA inhibitors had no effect on the SKF-38393-induced Ca2+ response. Taken together, our results suggest that the SKF-38393-induced Ca2+ response results from the direct activation of TRPV1 by a PLC/DAGmediated membrane-delimited pathway. These results provide evidence that the trans-activation of TRPV1 following D1R activation may contribute to the modulation of pain signaling in nociceptive sensory neurons. [ABSTRACT FROM AUTHOR]

Transplantation of neural stem cells (NSCs) has been proposed as a treatment for Parkinson disease (PD). The aim of this study was to monitor the viability of transplanted NSCs expressing the enhanced luciferase gene in a mouse model of PD in vivo, The PD animal model was induced by unilateral injection of 6-hydroxydopamine (6-OHDA). The behavioral test using apomorphine-induced rotation and positron emission tomography with [18F]N-(3-fluoropropyl)-2'-carbomethoxy-3'-(4-iodophenyl)nortropane ([18F]FP-CIT) were conducted. HB1.F3 cells transduced with an enhanced firefly luciferase retroviral vector (F3-effLuc cells) were transplanted into the right striatum. In vivo bioluminescence imaging was repeated for 2 weeks. Four weeks after transplantation, [18F]FP-CIT PET and the rotation test were repeated. All 6-OHDA-injected mice showed markedly decreased [18F]FP-CIT uptake in the right striatum. Transplanted F3-effLuc cells were visualized on the right side of the brain in all mice by bioluminescence imaging. The bioluminescence intensity of the transplanted F3-effLuc cells gradually decreased until it was undetectable by 10 days. The behavioral test showed that stem cell transplantation attenuated the motor symptoms of PD. No significant change was found in [18F]FP-CIT imaging after cell transplantation. We successfully established an in vivo bioluminescence imaging system for the detection of transplanted NSCs in a mouse model of PD. NSC transplantation induced behavioral improvement in PD model mice. [ABSTRACT FROM AUTHOR]

Abstract: Adiponectin is expressed in adipose tissue by adipogenic transcription factors including PPARγ, C/EBPα, and ADD1/SREBP1c. Because cAMP-response element binding protein (CREB) is also a central transcriptional activator of adipocyte differentiation, we evaluated CREB to determine if it stimulates adiponectin gene expression. To accomplish this, we evaluated the effects of activated CREB on the promoter activity of the mouse adiponectin gene, and identified the cAMP-response element (CRE) in the promoter. The constitutively active form of CREB increased the promoter activity of the mouse adiponectin gene. In addition, transfection studies using 5′ serial deleted promoters revealed the presence of a putative CRE located between the −1250 and −1000bp region. Furthermore, an electrophoresis mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP) analysis demonstrated that CREB bound to the region between −1022 and −995 in the adiponectin promoter. Insulin-like growth factor (IGF-1), which activate CREB, increased the adiponectin promoter activity. However, this stimulation was prevented by the dominant negative form of CREB (ACREB) and pretreatment with PD098059, indicating that IGF-1 stimulate adiponectin expression through CREB phosphorylation via the ERK pathway. Importantly, the transactivation of adiponectin expression by CREB was inhibited by ATF3. Coimmunoprecipitation and GST pull-down assay revealed that ATF3 bound to CREB and prevented CREB phosphorylation induced during differentiation of 3T3-L1 adipocytes. Collectively, these findings demonstrate that CREB is a positive regulator of mouse adiponectin gene expression in adipocytes, which play an important role in the regulation of adiponectin expression in response to growth factor. [Copyright &y& Elsevier]

Synaptic plasticity, the ability of synapses to change in strength, requires alterations in synaptic molecule compositions overtime, and synapses undergo selective modifications on stimulation. Molecular motors operate in sorting/transport of neuronal proteins; however, the targeting mechanisms that guide and direct cargo delivery remain elusive. We addressed the impact of synaptic transmission on the regulation of intracellular microtubule (MT)-based transport. We show that increased neuronal activity, as induced through GIyR activity blockade, facilitates tubulin polyglutamylation, a posttranslational modification thought to represent a molecular traffic sign for transport. Also, GIyR activity blockade alters the binding of the MT-associated protein MAP2 to MTs. By using the kinesin (KIF5) and the postsynaptic protein gephyrin as models, we show that such changes of MT tracks are accompanied by reduced motor protein mobility and cargo delivery into neurites. Notably, the observed neurite targeting deficits are prevented on functional depletion or gene expression knockdown of neuronal polyglutamylase. Our data suggest a previously undescribed concept of synaptic transmission regulating MT-dependent cargo delivery. [ABSTRACT FROM AUTHOR]

Effects of MK-801 (a NMDA receptor blocker) and CNQX (6-cyano-7-nitroquinoxaline-2,3-dione; a non-NMDA receptor blocker) on several neurotoxic responses induced by kainic acid (KA) were examined in ICR mice. In a lethality test, intracerebroventricular (i.c.v.) pretreatment of MK-801 (1 μg), but not CNQX (0.5 μg), attenuated the time to lethality induced by KA (0.5 μg)administered i.c.v. In the memory test (a passive avoidance test), MK-801, but not CNQX, prevented the memory loss induced by KA (0.1 μg). The damage induced by KA (0.1 μg)administered i.c.v. in the hippocampus was markedly concentrated in the CA3 pyramidal neurons. Both MK-801 and CNQX blocked the pyramidal cell death in CA3 hippocampal region induced by KA. In the immunocytochemical study, KA dramatically increased the phosphorylated ER (p-ERK) and decreased the phosphorylated CREB (p-CREB) in the hippocampus. Both MK-801 and CNQX attenuated, in part, the increased p-ERK and the decreased p-CREB induced by KA. In addition, both MK-801 and CNQX partially reduced the increased c-Fos and c-Jun protein expression in hippocampus induced by KA. Our results suggest that both NMDA and non-NMDA receptors are involved in supraspinally administered KA-induced pyramidal cell death in CA3 region of hippocampus in the mouse and the p-ERK and the dephosphorylation of CREB protein may play an important role in CA3 region cell death of the hippocampus induced by KA administered supraspinally. Furthermore, c-Fos and c-Jun proteins may serve as third messengers responsible for CA3 pyramidal cell death induced by supraspinally administered KA. [ABSTRACT FROM AUTHOR]

Background: We have previously reported that histamine-induced pruritus was attenuated in toll-like receptor 4 (TLR4) knockout mice due to decreased transient receptor potential V1 (TRPV1) sensitivity. Our results implied that TLR4 potentiated TRPV1 activation in sensory neurons; however, the molecular mechanism has yet to be elucidated. In this study, we investigated the molecular mechanisms of TLR4-mediated TRPV1 potentiation using TLR4-deficient sensory neurons and a heterologous expression system. Methods: Primary sensory neurons were obtained from wild-type or TLR4 knockout mice, and HEK293T cells expressing TRPV1 and TLR4 were prepared by transient transfection. TRPV1 activity was analyzed by calcium imaging, fluorophotometry, and patch-clamp recording. Subcellular protein distribution was tested by immunocytochemistry and cell surface biotinylation assay. Protein interaction was assessed by western blot and immunoprecipitation assay. Results: Direct association between TRPV1 and TLR4 was detected in HEK293T cells upon heterologous TRPV1 and TLR4 expression. In an immunoprecipitation assay using TLR4-deletion mutants and soluble toll/interleukin-1 receptor (TIR) protein, the cytoplasmic TIR domain of TLR4 was required for TLR4-TRPV1 association and TRPV1 potentiation. In TLR4-deficient sensory neurons, the activation-induced desensitization of TRPV1 increased, accompanied by enhanced TRPV1 clearance from the cell membrane upon activation compared to wild-type neurons. In addition, heterologous TLR4 expression inhibited activation-induced TRPV1 endocytosis and lysosomal degradation in HEK293T cells. Conclusion: Our data show that direct association between TRPV1 and TLR4 through the TIR domain enhances TRPV1 activity by blocking activation-induced TRPV1 desensitization. [ABSTRACT FROM AUTHOR]

Abstract: Neuronatin (Nnat) was initially identified as a selectively-expressed gene in neonatal brains, but its expression has been also identified in pancreatic β-cells. Therefore, to investigate the possible functions that Nnat may serve in pancreatic β-cells, two Nnat isotypes (α and β) were expressed using adenoviruses in murine MIN6N8 pancreatic β-cells, and the cellular fates and the effects of Nnat on insulin secretion, high glucose-induced apoptosis, and functional impairment were examined. Nnatα and Nnatβ were primarily localized in the endoplasmic reticulum (ER), and their expressions increased insulin secretion by increasing intracellular calcium levels. However, under chronic high glucose conditions, the Nnatβ to Nnatα ratio gradually increased in proportion to the length of exposure to high glucose levels. Moreover, adenovirally-expressed Nnatβ was inclined to form aggresome-like structures, and we found that Nnatβ aggregation inhibited the function of the proteasome. Therefore, when glucose is elevated, the expression of Nnatβ sensitizes MIN6N8 cells to high glucose stress, which in turn, causes ER stress. As a result, expression of Nnatβ increased hyperglycemia-induced apoptosis. In addition, the expression of Nnatβ under high glucose conditions decreased the expression of genes important for β-cell function, such as glucokinase (GCK), pancreas duodenum homeobox-1 (PDX-1), and insulin. Collectively, Nnat may play a critical factor in normal β-cell function, as well as in the pathogenesis of type 2 diabetes. [Copyright &y& Elsevier]

Aspirin (ASA) is a widely used oral analgesic which acts as an inhibitor of cyclooxygenase. Acetaminophen (ACET) is also an effective analgesic and may selectively inhibit brain prostaglandin synthetase. Various proinflammatory cytokines injected into the central nervous system show pain behavior. In the present study, the effects of orally administered ASA and ACET on pain behaviors induced by various proinflammatory cytokines were examined. At a dose of 100 mg/kg, ASA or ACET did not affect the pain behavior induced by TNF-α (100 pg), IL-1β (100 pg) or IFN-γ (100 pg) administered intrathecally. However, at doses of 200 and 300 mg/kg, ASA or ACET significantly and dose-dependently attenuated pain behavior induced by TNF-α, IL-1β or IFN-γ administered intrathecally. Our results suggest that orally administered ASA and ACET produce antinociception by inhibiting the nociceptive action of TNF-α, IL-1β or IFN-γ administered intrathecally. Copyright © 2005 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Platycodin D (PD), one of several triterpene saponins, was isolated from roots of Platycodon grandiflorum. We previously reported that intracerebroventricular (i.c.v.) administration of PD showed an antinociceptive effect as measured by the tail-flick assay. However, its exact role in the regulation of antinociception in the various types of pain models has not yet been characterized. Thus, we attempted to find antinociceptive profiles of PD in various pain models. PD administered intraperitoneally (i.p.), i.c.v. or intrathecally (i.t.) showed antinociceptive effects in dose-dependent manners as measured by the tail-flick, writhing and formalin tests. In the tail-flick test, PD at the low doses reached the peak after 15 minutes and returned to the control level after 60 minutes. However, higher doses of PD showed a strong antinociception at least for 1 hour. PD administered i.t. showed stronger antinociception than that induced by i.c.v. administration PD in both tail-flick and writhing tests. In the formalin test, PD administered i.p., i.c.v. or i.t. showed antinociceptive effects during both the first (direct nociceptive stimulation) and second (late inflammatory) phases. Pretreatment with naltrexone i.p., i.c.v. or i.t. did not affect PD-induced inhibition of the tail-flick response. Our results suggest that PD shows a strong antinociceptive effect on the tail-flick, writhing and formalin tests, acting on central nervous system. However, PD-induced antinociception may not be mediated by the opioid receptors. [ABSTRACT FROM AUTHOR]

Several studies have demonstrated that the nonselective opioid receptor antagonist naloxone produces a paradoxical antinociception in the formalin test. The opioid system is related to the serotonergic system for producing antinociception at the spinal level. Here we also asked whether systemic (i.p.) and intrathecal (i.t.) administrations of a nonselective serotonergic antagonist, methysergide, might produce paradoxical antinocicpetion similar to naloxone in the mouse formalin test. A diluted formalin solution was injected into the mouse plantar region of the hind paw and the duration of licking responses was measured at periods of 0-5 min (1st phase) and 20-40 min (2nd phase) after formalin injection. Methysergide administered i.p. and i.t. showed an attenuated licking duration only in the 2nd phase. The effect observed in the 2nd phase was reversed in the 5,7- dihydroxytriptamine, but not N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine pretreated group of mice, suggesting that descending serotonergic, but not noradrenergic, systems are involved in the methysergide antinociception. To further investigate the mechanism by which methysergide inhibited the nociceptive behaviors induced by formalin, the antinociceptive effect of methysergide was also tested in substance P (i.t.) and excitatory amino acids (i.t.), such as glutamate, N-methyl-D-aspartic acid, α-amino-3-hydroxy-5-methylisoxazole-4-propionic acid, and kainic acid, which are major components in the formalin-induced nociceptive transmission in the spinal cord pain models. The duration of nociceptive behaviors shown in these models was significantly shortened by i.p. and i.t. administration of methysergide. These results suggest that methysergide also produces a paradoxical antinociception in various pain models including the formalin test, similar to the results of naloxone. [ABSTRACT FROM AUTHOR]

COVID-19 exhibits extensive patient-to-patient heterogeneity. To link immune response variation to disease severity and outcome over time, we longitudinally assessed circulating proteins as well as 188 surface protein markers, transcriptome, and T cell receptor sequence simultaneously in single peripheral immune cells from COVID-19 patients. Conditional-independence network analysis revealed primary correlates of disease severity, including gene expression signatures of apoptosis in plasmacytoid dendritic cells and attenuated inflammation but increased fatty acid metabolism in CD56dimCD16hi NK cells linked positively to circulating interleukin (IL)-15. CD8+ T cell activation was apparent without signs of exhaustion. Although cellular inflammation was depressed in severe patients early after hospitalization, it became elevated by days 17–23 post symptom onset, suggestive of a late wave of inflammatory responses. Furthermore, circulating protein trajectories at this time were divergent between and predictive of recovery versus fatal outcomes. Our findings stress the importance of timing in the analysis, clinical monitoring, and therapeutic intervention of COVID-19. [Display omitted] • Multiparameter metric captures finer shades of disease severity to empower analyses • Network analysis reveals primary disease severity correlates in pDCs and NK cells • Analysis of timing effects suggests a late immune response juncture • Inflammatory divergence at the late juncture predicts fatal-recovery outcomes A quantitative disease severity metric is developed by integrating clinical data and circulating cytokines to capture finer shades of disease severity. Multimodal single-cell profiling and immune cell analyses of COVID-19 patients over time reveal a late wave of diverging inflammatory and host immune responses that predicts recovery versus fatality. [ABSTRACT FROM AUTHOR]