A Single Episode of Cortical Spreading Depolarization Increases mRNA Levels of Proinflammatory Cytokines,Calcitonin Gene-Related Peptide and Pannexin-1 Channels in the Cerebral Cortex

Cortical spreading depolarization (CSD) is the neuronal correlate of migraine aura and the reliable consequence of acute brain injury. The role of CSD in triggering headaches that follow migraine aura and brain injury remains to be uncertain. We examined whether a single CSD occurring in awake animals modified the expression of proinflammatory cytokines (Il1b, TNF, and Il6) and endogenous mediators of nociception/neuroinflammation-pannexin 1 (Panx1) channel and calcitonin gene-related peptide (CGRP), transforming growth factor beta (TGFb) in the cortex. Unilateral microinjury of the somatosensory cortex triggering a single CSD was produced in awake Wistar rats. Three hours later, tissue samples from the lesioned cortex, intact ipsilesional cortex invaded by CSD, and homologous areas of the contralateral sham-treated cortex were harvested and analyzed using qPCR. Three hours post-injury, intact CSD-exposed cortexes increased TNF, Il1b, Panx1, and CGRP mRNA levels. The strongest upregulation of proinflammatory cytokines was observed at the injury site, while CGRP and Panx1 were upregulated more strongly in the intact cortexes invaded by CSD. A single CSD is sufficient to produce low-grade parenchymal neuroinflammation with simultaneous overexpression of Panx1 and CGRP. The CSD-induced molecular changes may contribute to pathogenic mechanisms of migraine pain and post-injury headache.     

Sustained Effects of CGRP Blockade on Cortical Spreading Depolarization-Induced Alterations in Facial Heat Pain Threshold,Light Aversiveness,and Locomotive Activity in the Light Environment

A migraine is clinically characterized by repeated headache attacks that entail considerable disability. Many patients with migraines experience postdrome, the symptoms of which include tiredness and photophobia. Calcitonin gene-related peptide (GGRP) is critically implicated in migraine pathogenesis. Cortical spreading depolarization (CSD), the biological correlate of migraine aura, sensitizes the trigeminovascular system. In our previous study, CSD caused hypomotility in the light zone and tendency for photophobia at 72 h, at which time trigeminal sensitization had disappeared. We proposed that this CSD-induced disease state would be useful for exploring therapeutic strategies for migraine postdrome. In the present study, we observed that the CGRP receptor antagonist, olcegepant, prevented the hypomotility in the light zone and ameliorated light tolerability at 72 h after CSD induction. Moreover, olcegepant treatment significantly elevated the threshold for facial heat pain at 72 h after CSD. Our results raise the possibility that CGRP blockade may be efficacious in improving hypoactivity in the light environment by enhancing light tolerability during migraine postdrome. Moreover, our data suggest that the CGRP pathway may lower the facial heat pain threshold even in the absence of overt trigeminal sensitization, which provides an important clue to the potential mechanism whereby CGRP blockade confers migraine prophylaxis.     

TRPA1-Mediated Src Family Kinases Activity Facilitates Cortical Spreading Depression Susceptibility and Trigeminovascular System Sensitization

Transient receptor potential ankyrin 1 (TRPA1) plays a role in migraine and is proposed as a promising target for migraine therapy. However, TRPA1-induced signaling in migraine pathogenesis is poorly understood. In this study, we explored the hypothesis that Src family kinases (SFKs) transmit TRPA1 signaling in regulating cortical spreading depression (CSD), calcitonin gene-related peptide (CGRP) release and neuroinflammation. CSD was monitored in mouse brain slices via intrinsic optical imaging, and in rats using electrophysiology. CGRP level and IL-1β gene expression in mouse trigeminal ganglia (TG) was detected using Enzyme-linked Immunosorbent Assay and Quantitative Polymerase Chain Reaction respectively. The results showed a SFKs activator, pYEEI (EPQY(PO3H2)EEEIPIYL), reversed the reduced cortical susceptibility to CSD by an anti-TRPA1 antibody in mouse brain slices. Additionally, the increased cytosolic phosphorylated SFKs at Y416 induced by CSD in rat ipsilateral cerebral cortices was attenuated by pretreatment of the anti-TRPA1 antibody perfused into contralateral ventricles. In mouse TG, a SFKs inhibitor, saracatinib, restored the CGRP release and IL-1β mRNA level increased by a TRPA1 activator, umbellulone. Moreover, umbellulone promoted SFKs phosphorylation, which was reduced by a PKA inhibitor, PKI (14–22) Amide. These data reveal a novel mechanism of migraine pathogenesis by which TRPA1 transmits signaling to SFKs via PKA facilitating CSD susceptibility and trigeminovascular system sensitization.     

KCNK18 Biallelic Variants Associated with Intellectual Disability and Neurodevelopmental Disorders Alter TRESK Channel Activity

The TWIK-related spinal cord potassium channel (TRESK) is encoded by KCNK18, and variants in this gene have previously been associated with susceptibility to familial migraine with aura (MIM #613656). A single amino acid substitution in the same protein, p.Trp101Arg, has also been associated with intellectual disability (ID), opening the possibility that variants in this gene might be involved in different disorders. Here, we report the identification of KCNK18 biallelic missense variants (p.Tyr163Asp and p.Ser252Leu) in a family characterized by three siblings affected by mild-to-moderate ID, autism spectrum disorder (ASD) and other neurodevelopment-related features. Functional characterization of the variants alone or in combination showed impaired channel activity. Interestingly, Ser252 is an important regulatory site of TRESK, suggesting that alteration of this residue could lead to additive downstream effects. The functional relevance of these mutations and the observed co-segregation in all the affected members of the family expand the clinical variability associated with altered TRESK function and provide further insight into the relationship between altered function of this ion channel and human disease.     

Polymorphisms of the Proinflammatory Cytokine Genes Modulate the Response to NSAIDs but Not to Triptans in Migraine Attacks

Migraine is a common neurovascular disorder characterized by recurrent episodes of headache and associated neurological symptoms. At present, a significant portion of patients do not obtain a satisfactory response to acute pain-relieving therapies, including NSAIDs and triptans. In this context, pharmacogenetics plays a key role in the understanding of such a diverse response. In order to investigate whether functional polymorphisms in proinflammatory cytokine genes (IL-1α, IL-1β, IL-1RN; IL-6 and TNF-α) may influence the response to acute treatment, 313 consecutive patients with episodic migraine without aura were enrolled. Pain relief by administration of NSAIDs or triptans for three consecutive migraine attacks was evaluated. We found a significant association between A allele of the TNF-α promoter (−308 A/G) and a lack of efficacy after NSAID administration (p < 0.01, OR 2.51, 95% CI: 1.33 < OR < 4.75 compared to the G allele). Remaining polymorphisms had no significant effect on pain relief. Our study showed that a functional polymorphism in the TNF-α gene significantly modulates the clinical response to NSAID administration in acute attacks. Patients with higher production of the active cytokine during stress showed a significantly lower anti-migraine effect. Our results further support a role for TNF-α in the pathophysiological mechanisms of migraine attack.     

The HP1a disordered C terminus and chromo shadow domain cooperate to select target peptide partners

Drosophila melanogaster heterochromatin protein 1a (HP1a) is essential for compacted heterochromatin structure and the associated gene silencing. Its chromo shadow domain (CSD) is well known for binding to peptides that contain a PXVXL motif. Heterochromatin protein 2 (HP2) is a non-histone chromosomal protein that associates with HP1a in the pericentric heterochromatin, telomeres, and the fourth chromosome. Using NMR spectroscopy, fluorescence polarization, and site-directed mutagenesis, we identified an LCVKI motif in HP2 that binds to the HP1a CSD. The binding affinity of the HP2 fragment is approximately two orders of magnitude higher than that of peptides from PIWI (with a PRVKV motif), AF10 (with a PLVVL motif), or CG15356 (with LYPLL and LSIVA motifs). To delineate differential interactions of the HP1a CSD, we characterized its structure, backbone dynamics, and dimerization constant. We found that the dimerization constant is bracketed by the affinities of HP2 and PIWI, which dock to the same HP1a homodimer surface. This suggests that HP2, but not PIWI, interaction can drive the homodimerization of HP1a. Interestingly, the integrity of the disordered C-terminal extension (CTE) of HP1a is essential for discriminatory binding, whereas swapping the PXVXL motifs does not confer specificity. Serine phosphorylation at the peptide binding surface of the CSD is thought to regulate heterochromatin assembly. Glutamic acid substitution at these sites destabilizes HP1a dimers, but improves the interaction with both binding partners. Our studies underscore the importance of CSD dimerization and cooperation with the CTE in forming distinct complexes of HP1a.     

CFD-PBE simulation of premixed continuous precipitation incorporating nucleation,growth and aggregation in a stirred tank with multi-class method

The continuous precipitation of BaSO₄ with a premixed feed in a stirred tank is simulated. The population balance equation (PBE) is closed with the multi-class method. The effects of aggregation, feeding concentration, agitation speed, mean residence time, the crystal size distribution (CSD) at different locations and the net birth rate of each size bin due to aggregation are numerically studied. Size bin number tests show that at least 36 bins are needed for the size range in this work to achieve acceptable accuracy in both cases with and without aggregation. The predicted CSD is in good agreement with the reported experimental measurement. Narrower CSD and higher peak value are obtained with higher feed concentration. Aggregation broadens the CSD, decreases the peak value, and makes the CSD shift toward a larger size. The mean size L₃₂ of crystals decreases significantly with increasing the inlet concentration, which is consistent with the experiments and model predictions reported in the literature. The effects of residence time and agitation speed on CSD and L₃₂ are limited, which is very different from those in unpremixed cases. The CSD at the inlet region differs greatly from the outlet and near impeller regions, both in shape and value. A bimodal distribution appears around the inlet under some conditions. The net birth rate of the ith size bin due to aggregation increases dramatically with the inlet concentration. The detailed information, such as the bimodal distribution in a certain location under certain conditions and the aggregation rate of each size bin, could be useful for the precise and sophisticated control of precipitation in some novel processes like nano-particles production.     

相对分子质量分布对双峰聚乙烯薄膜树脂性能的影响

通过差示扫描量热仪、高温凝胶渗透色谱仪、旋转流变仪表征了2种具有不同力学性能的双峰高密度聚乙烯薄膜树脂（FHM 9455F1-A、FHM 9455F1-B）结构的差异性,FHM 9455F1-A比 FHM 9455F1-B具有更高的冲击强度和拉伸强度。结果表明,FHM 9455F1-A高相对分子质量部分的相对含量更大,片晶间能够形成更多的系带分子,改善了材料的冲击强度,而片晶厚度,特别是厚片晶部分的片晶厚度更厚,使得材料具有更高的拉伸强度。     

Association of LTA and SOD Gene Polymorphisms with Cerebral White Matter Hyperintensities in Migraine Patients

White matter hyperintensities (WMHs) in migraine could be related to inflammatory and antioxidant events. The aim of this study is to verify whether migraine patients with WMHs carry a genetic pro-inflammatory/pro-oxidative status. To test this hypothesis, we analyzed lymphotoxin alpha (LTA; rs2071590T and rs2844482G) and superoxide dismutase 1 (SOD1; rs2234694C) and 2 (SOD2; rs4880T) gene polymorphisms (SNPs) in 370 consecutive patients affected by episodic (EM; n = 251) and chronic (CM; n = 119) migraine and in unrelated healthy controls (n = 100). Brain magnetic resonance was available in 183/370 patients. The results obtained show that genotypes and allele frequencies for all tested SNPs did not differ between patients and controls. No association was found between single SNPs or haplotypes and sex, migraine type, cardiovascular risk factors or disorders. Conversely, the LTA rs2071590T (OR = 2.2) and the SOD1 rs2234694C (OR = 4.9) alleles were both associated with WMHs. A four-loci haplotype (TGCT haplotype: rs2071590T/rs2844482G/rs2234694C/rs4880T) was significantly more frequent in migraineurs with WMHs (7 of 38) compared to those without WMHs (4 of 134; OR = 8.7). We may, therefore, conclude by suggesting that that an imbalance between pro-inflammatory/pro-oxidative and antioxidant events in genetically predisposed individuals may influence the development of WMHs.     

Enhanced cGMP Interactor Rap Guanine Exchange Factor 4 (EPAC2) Expression and Activity in Degenerating Photoreceptors: A Neuroprotective Response?

The disease retinitis pigmentosa (RP) leads to photoreceptor degeneration by a yet undefined mechanism(s). In several RP mouse models (i.e., rd mice), a high cyclic GMP (cGMP) level within photoreceptors is detected, suggesting that cGMP plays a role in degeneration. The rap guanine exchange factor 4 (EPAC2) is activated by cyclic AMP (cAMP) and is an accepted cGMP-interacting protein. It is unclear whether and how cGMP interacts with EPAC2 in degenerating photoreceptors; we therefore investigated EPAC2 expression and interactions with cGMP and cAMP in retinas of the rd1 and rd10 models for retinal degeneration. EPAC2 expression in the photoreceptor layer increased significantly during rd1 and rd10 degeneration, and an increase in EPAC2 interactions with cGMP but not cAMP in the rd1 was also seen via a proximity ligation assay on histological sections. Retinal explant cultures revealed that pharmacological inhibition of the EPAC2 activity reduced the photoreceptor layer thickness in the rd10 retina, suggesting that EPAC2 inhibition promotes degeneration. Taken together, our results support the hypothesis that high degeneration-related cGMP leads to increased EPAC2 and cGMP interactions, inhibiting EPAC2. By inference, EPAC2 could have neuroprotective capacities that may be exploited in the future.     

Simulation of reactive precipitation processes using the network-of-zones model

A 2-D network-of-zones model is extended and applied to a reactive precipitation process in batch mode. The simulations are performed for a network of size 2 × (10 × 10) for an elementary reaction through the solution of 1400 ODEs. The complicated interactions between mixing efficacy and the system kinetics are systematically investigated. When the stirrer speed is very slow, the crystal size distribution (CSD) of the product in the precipitator is determined by the intensity of mixing. Conversely, at higher stirrer speed, the CSD is controlled by the system kinetics. More effective mixing leads to an increase in the number of crystals, a reduction of the average size and a narrower crystal size distribution. The extended network-of-zones model presented in this work can be used conveniently for integrating computational fluid dynamics and reactive precipitation processes.     

Determination of crystal size distributions in alumina ceramics by a novel X‐ray diffraction procedure

A novel X‐ray diffraction based method is presented, capable of determining volume‐based crystal size distribution (CSD) of polycrystalline materials and crystalline powders with unprecedented sampling statistics; the method is named fast X‐ray diffraction crystal size distribution analysis (FXD‐CSD). FXD‐CSD can be performed with standard laboratory X‐ray diffractometers equipped with a position sensitive detector and uses a software package written in Python for the data analysis. FXD‐CSD is a destruction‐free and generally applicable method to establish CSDs of polycrystalline materials as well as powders for sizes well below 1 μm up to about 100 μm; it even allows for studies of samples enclosed in complex environments, e.g., for in situ measurements in a furnace or in a pressure cell. To show the capability of the method the microstructural evolution of four alumina substrates with different time‐spans of sintering (4, 8, 16, and 24 hour at 1600°C) is investigated via FXD‐CSD and SEM imaging. The corresponding CSDs and average grain sizes are determined, results obtained by FXD‐CSD and the line‐intersection methods are compared and clear evidence for the presence of abnormal grain growth (AGG) during sintering is shown. From three tested probability density functions (PDF) describing the CSDs a log‐normal PDF fits best to the volume based CSDs; the method provides size distributions with unprecedented precision opening the way to a systematic and meaningful comparison between theoretically predicted and observed CSDs.     

Preparation for graft copolymers of sawdust/vinyl monomers and their application for removal of dye

A kind of cationic exchanging resin of carboxyl sawdust (CSD) was fabricated through the hydrolysis of graft copolymers of sawdust with acrylonitrile (SAN) and sawdust with acrylamide (SAA) that were made by initiator Fe²⁺/H₂O₂. A study of the graft copolymerization was conducted for initiator usage, vinyl monomer usage, and reaction temperature. The hydrolysis under basic/acid conditions was also studied for the yield and acid value of CSD followed to adsorb Basic Pink dye (BPD). Our results show the following: (1) graft copolymers (SAN and SAA) with a high rate of graft copolymerization are readily prepared by suitable usages of initiator and vinyl monomer under a certain temperature; (2) adsorption capacity of CSD is relative to pH of BPD solution and reaches the most adsorption capacity at pH ≈ 6; (3) adsorption capacity of CSD increases along with the augment of its acid value; and (4) the adsorption capacity of the CSD toward BPD increases along with augment of initial concentration of the adsorbate and reaches about 500 mg/g. © 2002 Wiley Periodicals, Inc. J Appl Polym Sci 83: 2390–2396, 2002     

Low-temperature solution processing route for potassium sodium niobate (KNN) thin films

A major challenge for integration of functional oxides, such as ferroelectrics, into microelectronics and flexible electronics is the reduction of their processing temperature, which needs to be lower than the degradation temperature of silicon and flexible plastic substrates. Aiming that, attention has been given to low-temperature processing of oxide films by chemical solution deposition (CSD). In the field of ferroelectrics, potassium sodium niobate ((K_1−xNa_x)NbO₃, KNN) has aroused a special interest due to its eco-friendliness, despite the high crystallization temperature. In this work, polycrystalline KNN thin films have been fabricated for the first time at a temperature as low as 400 °C using a modified CSD process, the Seeded Photosensitive Precursor Method (SPPM). Despite this low processing temperature, monophasic and stoichiometric films with appreciable values of remnant polarization, P_r ∼ 10.8 μC/cm², and hysteretic piezoresponse are obtained. These results open a window to the direct integration of KNN films into the high-performance electronic devices.     

Modeling of cluster structure-dependent drag with Eulerian approach for circulating fluidized beds

Flow behavior of gas and solids is simulated in combination the gas-solid two-fluid model with a cluster structure-dependent (CSD) drag coefficient model. The dispersed phase is modeled by a Eulerian approach based upon the kinetic theory of granular flow (KTGF) including models for describing the dispersed phase interactions with the continuous phase. The drag forces of gas-solid phases are predicted from the local structure parameters of the dense and dilute phases based on the minimization of the energy consumed by heterogeneous drag. The cluster structure-dependent (CSD) drag coefficients are incorporated into the two-fluid model to simulate flow behavior of gas and particles in a riser. Simulation results indicate that the dynamic formation and dissolution of clusters can be captured with the cluster structure-dependent drag coefficient model. Simulated solid velocity and concentration of particles profiles are in reasonable agreement with experimental results.     

New Insights on Metabolic and Genetic Basis of Migraine: Novel Impact on Management and Therapeutical Approach

Migraine is a hereditary disease, usually one-sided, sometimes bilateral. It is characterized by moderate to severe pain, which worsens with physical activity and may be associated with nausea and vomiting, may be accompanied by photophobia and phonophobia. The disorder can occur at any time of the day and can last from 4 to 72 h, with and without aura. The pathogenic mechanism is unclear, but extensive preclinical and clinical studies are ongoing. According to electrophysiology and imaging studies, many brain areas are involved, such as cerebral cortex, thalamus, hypothalamus, and brainstem. The activation of the trigeminovascular system has a key role in the headache phase. There also appears to be a genetic basis behind the development of migraine. Numerous alterations have been identified, and in addition to the genetic cause, there is also a close association with the surrounding environment, as if on the one hand, the genetic alterations may be responsible for the onset of migraine, on the other, the environmental factors seem to be more strongly associated with exacerbations. This review is an analysis of neurophysiological mechanisms, neuropeptide activity, and genetic alterations that play a fundamental role in choosing the best therapeutic strategy. To date, the goal is to create a therapy that is as personalized as possible, and for this reason, steps forward have been made in the pharmacological field in order to identify new therapeutic strategies for both acute treatment and prophylaxis.     

Integrated Bioinformatics,Environmental Epidemiologic and Genomic Approaches to Identify Environmental and Molecular Links between Endometriosis and Breast Cancer

We present a combined environmental epidemiologic, genomic, and bioinformatics approach to identify: exposure of environmental chemicals with estrogenic activity; epidemiologic association between endocrine disrupting chemical (EDC) and health effects, such as, breast cancer or endometriosis; and gene-EDC interactions and disease associations. Human exposure measurement and modeling confirmed estrogenic activity of three selected class of environmental chemicals, polychlorinated biphenyls (PCBs), bisphenols (BPs), and phthalates. Meta-analysis showed that PCBs exposure, not Bisphenol A (BPA) and phthalates, increased the summary odds ratio for breast cancer and endometriosis. Bioinformatics analysis of gene-EDC interactions and disease associations identified several hundred genes that were altered by exposure to PCBs, phthalate or BPA. EDCs-modified genes in breast neoplasms and endometriosis are part of steroid hormone signaling and inflammation pathways. All three EDCs–PCB 153, phthalates, and BPA influenced five common genes—CYP19A1, EGFR, ESR2, FOS, and IGF1—in breast cancer as well as in endometriosis. These genes are environmentally and estrogen responsive, altered in human breast and uterine tumors and endometriosis lesions, and part of Mitogen Activated Protein Kinase (MAPK) signaling pathways in cancer. Our findings suggest that breast cancer and endometriosis share some common environmental and molecular risk factors.     

Leptosphaeria maculans-Brassica napus Battle: A Comparison of Incompatible vs. Compatible Interactions Using Dual RNASeq

Leptosphaeria maculans causes blackleg disease, which is one of the most destructive diseases of canola (Brassica napus L.). Due to the erosion of the current resistance in B. napus, it is pivotal to introduce new resistant genotypes to the growers. This study evaluated the potential of Rlm7 gene as resistance to its corresponding avirulence AvrLm7 gene is abundant. The Rlm7 line was inoculated with L. maculans isolate with AvrLm7; UMAvr7; and the CRISPR/Cas9 knockout AvrLm7 mutant, umavr7, of the same isolate to cause incompatible and compatible interactions, respectively. Dual RNA-seq showed differential gene expressions in both interactions. High expressions of virulence-related pathogen genes-CAZymes, merops, and effector proteins after 7-dpi in compatible interactions but not in incompatible interaction—confirmed that the pathogen was actively virulent only in compatible interactions. Salicyclic and jasmonic acid biosynthesis and signaling-related genes, defense-related PR1 gene (GSBRNA2T00150001001), and GSBRNA2T00068522001 in the NLR gene family were upregulated starting as early as 1- and 3-dpi in the incompatible interaction and the high upregulation of those genes after 7-dpi in compatible interactions confirmed the early recognition of the pathogen by the host and control it by early activation of host defense mechanisms in the incompatible interaction.     

Interplay between Ca2+/Calmodulin-Mediated Signaling and AtSR1/CAMTA3 during Increased Temperature Resulting in Compromised Immune Response in Plants

Changing temperatures are known to affect plant–microbe interactions; however, the molecular mechanism involved in plant disease resistance is not well understood. Here, we report the effects of a moderate change in temperature on plant immune response through Ca²⁺/calmodulin-mediated signaling. At 30 °C, Pst DC3000 triggered significantly weak and relatively slow Ca²⁺ influx in plant cells, as compared to that at 18 °C. Increased temperature contributed to an enhanced disease susceptibility in plants; the enhanced disease susceptibility is the result of the compromised stomatal closure induced by pathogens at high temperature. A Ca²⁺ receptor, AtSR1, contributes to the decreased plant immunity at high temperatures and the calmodulin-binding domain (CaMBD) is required for its function. Furthermore, both salicylic acid biosynthesis (ICS) and salicylic acid receptor (NPR1) are involved in this process. In addition to stomatal control, AtSR1 is involved in high temperature-compromised apoplastic immune response through the salicylic acid signaling pathway. The qRT-PCR data revealed that AtSR1 contributed to increased temperatures-mediated susceptible immune response by regulating SA-related genes in atsr1, such as PR1, ICS1, NPR1, as well as EDS1. Our results indicate that Ca²⁺ signaling has broad effects on the molecular interplay between changing temperatures as well as plant defense during plant–pathogen interactions.