Antinociceptive actions of honokiol and magnolol on glutamatergic and inflammatory pain

The antinociceptive effects of honokiol and magnolol, two major bioactive constituents of the bark of Magnolia officinalis, were investigated on animal paw licking responses and thermal hyperalgesia induced by glutamate receptor agonists including glutamate, N-methyl-D-aspartate (NMDA), and metabotropic glutamate 5 receptor (mGluR5) activator (RS)-2-chloro-5-hydroxyphenylglycine (CHPG), as well as inflammatory mediators such as substance P and prostaglandin E2 (PGE2) in mice. The actions of honokiol and magnolol on glutamate-induced c-Fos expression in the spinal cord dorsal horn were also examined. Our data showed that honokiol and magnolol blocked glutamate-, substance P- and PGE2-induced inflammatory pain with similar potency and efficacy. Consistently, honokiol and magnolol significantly decreased glutamate-induced c-Fos protein expression in superficial (I-II) laminae of the L4-L5 lumbar dorsal horn. However, honokiol was more selective than magnolol for inhibition of NMDA-induced licking behavioral and thermal hyperalgesia. In contrast, magnolol was more potent to block CHPG-mediated thermal hyperalgesia. These results demonstrate that honokiol and magnolol effectively decreased the inflammatory pain. Furthermore, their different potency on inhibition of nociception provoked by NMDA receptor and mGluR5 activation should be considered

Optimization of an Anti-NMDA Receptor Autoantibody Diagnostic Bioassay

Anti-N-methyl-D-aspartate receptor (anti-NMDAR) encephalitis is one of the most frequently encountered autoimmune encephalitis. The pathogenesis of both anti-NMDAR encephalitis and schizophrenia involve down-regulation of NMDA receptors. Whether autoantibody-mediated destruction of neuronal NMDA receptors is associated with schizophrenia or first-episode psychosis (FEP) remains unclear, as the current findings from different groups are inconsistent. The main culprits are likely due to heterogeneity of autoantibodies (autoAbs) in a patient's blood or cerebrospinal fluid (CSF), as well as due to limitation of the current detection methods for anti-NMDAR autoAbs. Here, we optimized the current diagnostic method based on the only commercially-available anti-NMDAR test kit. We first increased detection sensitivity by replacing reporter fluorophore fluorescein isothiocyanate (FITC) in the kit with Alexa Fluor 488, which is superior in resisting photobleaching. We also found that using an advanced imaging system could increase the detection limit, compared to using a simple fluorescence microscope. To improve test accuracy, we implemented secondary labeling with a well-characterized mouse anti-NR1 monoclonal antibody (mAb) after immunostaining with a patient's sample. The degree of colocalization between mouse and human antisera in NMDAR-expressing cells served to validate test results to be truly anti-NMDAR positive or false-positive. We also incorporated DNA-specific DAPI to simultaneously differentiate autoAbs targeting the plasma membrane from those targeting cell nuclei or perinuclear compartments. All the technical implementation could be integrated in a general hospital laboratory setting, without the need of specialized expertise or equipment. By sharing our experience, we hope this may help improve sensitivity and accuracy of the mainstream method for anti-NMDAR detection

Deep-sea water containing selenium provides intestinal protection against duodenal ulcers through the upregulation of Bcl-2 and thioredoxin reductase 1.

Deep-sea water (DSW), which is rich in micronutrients and minerals and with antioxidant and anti-inflammatory qualities, may be developed as marine drugs to provide intestinal protection against duodenal ulcers. We determined several characteristics in the modified DSW. We explored duodenal pressure, oxygenation, microvascular blood flow, and changes in pH and oxidative redox potential (ORP) values within the stomach and duodenum in response to tap water (TW, hardness: 2.48 ppm), DSW600 (hardness: 600 ppm), and DSW1200 (hardness: 1200 ppm) in Wistar rats and analyzed oxidative stress and apoptosis gene expressions by cDNA and RNA microarrays in the duodenal epithelium. We compared the effects of drinking DSW, MgCl2, and selenium water on duodenal ulcers using pathologic scoring, immunohistochemical analysis, and Western blotting. Our results showed DSW has a higher pH value, lower ORP value, higher scavenging H2O2 and HOCl activity, higher Mg2+ concentrations, and micronutrients selenium compared with TW samples. Water infusion significantly increased intestinal pressure, O2 levels, and microvascular blood flow in DSW and TW groups. Microarray showed DSW600, DSW1200, selenium water upregulated antioxidant and anti-apoptotic genes and downregulated pro-apoptotic gene expression compared with the TW group. Drinking DSW600, DSW1200, and selenium water but not Mg2+ water significantly enhanced Bcl-2 and thioredoxin reductase 1 expression. Bax/Bcl-2/caspase 3/poly-(ADP-ribose)-polymerase signaling was activated during the pathogenesis of duodenal ulceration. DSW drinking reduced ulcer area as well as apoptotic signaling in acetic acid-induced duodenal ulcers. DSW, which contains selenium, provides intestinal protection against duodenal ulcers through the upregulation of Bcl-2 and thioredoxin reductase 1

Additional file 2: Figures S2. of Effects of sarcosine and N, N-dimethylglycine on NMDA receptor-mediated excitatory field potentials

Effects of sarcosine or DMG combined with NMDA and NMDA plus glycine on excitatory field potentials. The effects of glutamate (100 μM) plus glycine (10 μM) and NMDA (30 μM) plus glycine (10 μM) was compared first and blockade by MK-801. The experimental protocol (A) and the representative EFPs recordings (B) were shown as application of sarcosine (100 μM) or DMG (100 μM) combined with NMDA or NMDA plus glycine. Sarcosine and DMG produced the same effects when glutamate was replaced by NMDA. (PDF 251 kb

Involvement of Pachybasin and Emodin in Self-Regulation of <i>Trichoderma harzianum</i> Mycoparasitic Coiling

Our aim was to determine the effects of two secondary metabolites secreted by <i>Trichoderma harzianum</i>, pachybasin and emodin, on the mycoparasitic coiling behavior and cAMP content of <i>T</i>. <i>harzianum</i>. The number of <i>T</i>. <i>harzianum</i> coils around Nylon 66 fiber was increased in the presence of <i>R</i>. <i>solani</i>. The number of <i>T</i>. <i>harzianum</i> coils around <i>R</i>. <i>solani</i> hyphae and Nylon 66 fiber were significantly increased in the presence of pachybasin and emodin. The cAMP level in <i>T</i>. <i>harzianum</i> was significantly increased by close contact with <i>R</i>. <i>solani</i> and much higer cAMP level in the presence of exogenous pachybasin and emodin. A cAMP inhibitor diminished the effect of pachybasin and emodin on <i>T</i>. <i>harzianum</i> coiling around Nylon 66 fiber. The results suggest that pachybasin and emodin mediate the increase in the number of <i>Trichoderma</i> mycoparasitic coils via cAMP signaling. This is the first report to suggest that pachybasin and emodin play roles in the biocontrol mechanism of <i>Trichoderma</i>

Additional file 1: Figure S1. of Effects of sarcosine and N, N-dimethylglycine on NMDA receptor-mediated excitatory field potentials

Characterization of DMG as a NMDA receptor glycine binding site partial agonist. The experimental protocol (AC) and the representative field potential recordings (BD) were shown. Increased concentration of glycine from 10 to 100 μM could surmount the inhibitory effect of DMG (100 μM). Elevated DMG concentration from 100 to 300 μM could attenuate the inhibitory effect of glycine binding site antagonist 7-CK (1 μM). (PDF 282 kb

Expression of apoptotic genes in the duodenal epithelium after water restriction, intake of DSW waters vs intake of TW.

<p><b>Fold-Change</b> [2∧(−Delta Delta Ct)] is the normalized gene expression [2∧(−Delta Ct)] in the test sample divided by the normalized gene expression [2∧(−Delta Ct)] in the control sample.</p

Effect of water infusion on intestinal pressure, microvascular blood flow and oxygen concentration.

<p>The typical recordings (A) and statistical data (B) relating to changes in arterial blood pressure (ABP), intestinal pressure (IP), duodenal microvascular blood flow (DMVBF) and partial oxygen concentration in the duodenum in response to tap water (TW) or deep-sea water (DSW) infusion. C = control stage; I = intestinal infusion of TW or DSW; R = recovery stage after infusion. *<i>P</i><0.05 vs. control stage.</p