Contextual control of conditioned pain tolerance and endogenous analgesic systems: Evidence for sex-based differences in endogenous opioid engagement

The mechanisms underlying the transition from acute to chronic pain are unclear but may involve the persistence or strengthening of pain memories acquired in part through associative learning. Contextual cues, which comprise the surrounding environment where events occur, were recently described as a critical regulator of pain memory; both rodents and humans exhibit increased pain sensitivity in environments recently associated with a single painful experience. It is unknown, however, how repeated exposure to an acute painful unconditioned stimulus in a distinct context modifies pain sensitivity or the expectation of pain in that environment. To answer this question, we conditioned mice to associate distinct contexts with either repeated administration of a mild visceral pain stimulus (intraperitoneal injection of acetic acid) or vehicle injection over the course of three days. On the final day of experiments animals received either an acid injection or vehicle injection prior to being placed into both contexts. In this way, contextual control of pain sensitivity and pain expectation could be tested respectively. Both male and female mice developed context-dependent conditional pain tolerance, a phenomenon mediated by endogenous opioid signaling. However, when expecting the presentation of a painful stimulus in a given context, males exhibited conditional hypersensitivity whereas females exhibited endogenous opioid-mediated conditional analgesia. Successful determination of the brain circuits involved in this sexually dimorphic anticipatory response may allow for the manipulation of pain memories, which may contribute to the development of chronic pain states.

Dual Inhibition of FAAH and MAGL Counteracts Migraine-like Pain and Behavior in an Animal Model of Migraine

The endocannabinoid system exerts an important role in pain processing and modulation. Modulation of the system with hydrolase inhibitors of anandamide (AEA) or 2-arachidonyl glycerol (2-AG) has proved effective in reducing migraine-like features in animal models of migraine. Here, we investigated the effect of dual inhibition of the AEA and 2-AG catabolic pathways in the nitroglycerin-based animal model of migraine. The dual inhibitor JZL195 was administered to rats 2 h after nitroglycerin or vehicle injection. Rats were then exposed to the open field test and the orofacial formalin test. At the end of the tests, they were sacrificed to evaluate calcitonin gene-related peptide (CGRP) serum levels and gene expression of CGRP and cytokines in the cervical spinal cord and the trigeminal ganglion. The dual inhibitor significantly reduced the nitroglycerin-induced trigeminal hyperalgesia and pain-associated behavior, possibly via cannabinoid 1 receptors-mediated action, but it did not change the hypomotility and the anxiety behaviors induced by nitroglycerin. The decreased hyperalgesia was associated with a reduction in CGRP and cytokine gene expression levels in central and peripheral structures and reduced CGRP serum levels. These data suggest an antinociceptive synergy of the endocannabinoid action in peripheral and central sites, confirming that this system participates in reduction of cephalic pain signals.

Blocking N-Methyl-D-aspartic Acid (NMDA) Receptor Inhibits Heat-Sensitization Response of Moxibustion in Stroke Rats

Our previous studies demonstrated that effects of moxibustion heavily relied on heat-sensitization response, a specific sensation induced by moxibustion in the ill body. On the sensation, long-term potentiation (LTP) of prelimbic cortex was attributed to heat-sensitization responses. The N-methyl-D-aspartic acid (NMDA) receptor plays a key role in LTP induction; however, little is known about the role of NMDA receptor in heat-sensitization response. The present study investigated the role of NMDA receptor in heat-sensitization response, specifically, NMDA receptor was inhibited by competitive glutamatergic antagonist, (±)-3-(2-carboxypiperazin-4-yl)propyl-1-phosphonic acid (CPP), observing the frequency of heat-sensitization response in moxibustion treatment and evaluating the conducive outcomes to cerebral infarct rats for rehabilitation. Heat-sensitization response in cerebral infarct rats was regularly measured for all the samples when exposed to moxibustion. Intraperitoneal injection of CPP was conducted, and soon afterwards, a significant drop of heat-sensitization response in all the samples was measured. Moreover, moxibustion efficiency on rehabilitation was unfavourably affected in cerebral infarct rats when compared to vehicle injection control. This indicated that NMDA receptor antagonist made a negative impact on induction of heat-sensitization response and consequently affected cerebral infarct rats to rehabilitate under moxibustion treatment. It also suggested that activating NMDA receptor played a positive part in ischemic stroke rehabilitation, and regulating its activity could be a feasible way to increase heat-sensitization response, improving the effect of moxibustion.

Early Changes in Crayfish Hemocyte Proteins after Injection with a ß-1,3-Glucan, Compared to Saline Injected and Naive Animals

Early changes in hemocyte proteins in freshwater crayfish Pacifastacus leniusculus, in response to an injection with the fungal pattern recognition protein β-1,3-glucan (laminarin) were investigated, as well as changes after saline (vehicle) injection and in naïve animals. Injection of saline resulted in rapid recruitment of granular hemocytes from surrounding tissues, whereas laminarin injection on the other hand induced an initial dramatic drop of hemocytes. At six hours after injection, the hemocyte populations therefore were of different composition. The results show that mature granular hemocytes increase in number after saline injection as indicated by the high abundance of proteins present in granular cell vesicles, such as a vitelline membrane outer layer protein 1 homolog, mannose-binding lectin, masquerade, crustin 1 and serine protease homolog 1. After injection with the β-1,3-glucan, only three proteins were enhanced in expression, in comparison with saline-injected animals and uninjected controls. All of them may be associated with immune responses, such as a new and previously undescribed Kazal proteinase inhibitor. One interesting observation was that the clotting protein was increased dramatically in most of the animals injected with laminarin. The number of significantly affected proteins was very few after a laminarin injection when compared to uninjected and saline-injected crayfish. This finding may demonstrate some problematic issues with gene and protein expression studies from other crustaceans receiving injections with pathogens or pattern recognition proteins. If no uninjected controls are included and no information about hemocyte count (total or differential) is given, expressions data for proteins or mRNAs are very difficult to properly interpret.

Secreted Frizzled Protein 3 Is A Novel Cardioprotective Mechanism Unique to the Clinically Relevant 4th Window of Ischemic Preconditioning

Most studies on ischemic preconditioning (IPC) use one or two ischemic stimuli, before examining cardioprotection. In order to better simulate the clinical situation, we examined, in pigs, the effects of 6 episodes of 10 min coronary artery occlusion (CAO) 12 hours apart, followed by 60 min CAO. We named this model the 4th window of IPC. In order to determine the novel mechanisms mediating cardioprotection in the 4th window, gene analysis was examined in 4th window IPC cardiac tissue, 60 min after the last episode of 10 min CAO. Secreted Frizzled Related Protein 3 (sFRP3) was the most significantly upregulated gene that was unique to the 4th window, i.e., not found in the 1st, 2nd or 3rd window IPC. To study the effects of sFRP3 on cardioprotection, sFRP3 was injected in the hearts of WT mice. In the CAO/CAR model (30 min CAO followed by 24 hour CAR), infarct size was less, p<0.01, after sFRP3 injection (14±1.7%) compared to vehicle injection (48±1.6%). sFRP3 injection also protected the development of heart failure following permanent CAO for 2 wks. Left ventricular ejection fraction was significantly improved, p<0.05, at 2 weeks after CAO with sFRP3 (53±5%) compared with vehicle (36±2%), and was accompanied by significant, p<0.01, reductions in myocardial fibrosis (53±4%), myocyte size (17±3%), apoptosis (100%) and mortality (56%). Thus, sFRP3, unique to the clinically relevant 4th window IPC model, is a novel mechanism mediating ischemic cardioprotection.

Blockade of Acid-Sensing Ion Channels Increases Urinary Bladder Capacity With or Without Intravesical Irritation in Mice

We conducted this study to examine whether acid-sensing ion channels (ASICs) are involved in the modulation of urinary bladder activity with or without intravesical irritation induced by acetic acid. All in vivo evaluations were conducted during continuous infusion cystometry in decerebrated unanesthetized female mice. During cystometry with a pH 6.3 saline infusion, an i.p. injection of 30 μmol/kg A-317567 (a potent, non-amiloride ASIC blocker) increased the intercontraction interval (ICI) by 30% (P &lt; 0.001), whereas vehicle injection had no effect. An intravesical acetic acid (pH 3.0) infusion induced bladder hyperactivity, with reductions in ICI and maximal voiding pressure (MVP) by 79% (P &lt; 0.0001) and 29% (P &lt; 0.001), respectively. A-317567 (30 μmol/kg i.p.) alleviated hyperreflexia by increasing the acid-shortened ICI by 76% (P &lt; 0.001). This dose produced no effect on MVP under either intravesical pH condition. Further analysis in comparison with vehicle showed that the increase in ICI (or bladder capacity) by the drug was not dependent on bladder compliance. Meanwhile, intravesical perfusion of A-317567 (100 μM) had no effect on bladder activity during pH 6.0 saline infusion cystometry, and drug perfusion at neither 100 μM nor 1 mM produced any effects on bladder hyperreflexia during pH 3.0 acetic acid infusion cystometry. A-317567 has been suggested to display extremely poor penetrability into the central nervous system and thus to be a peripherally active blocker. Taken together, our results suggest that blockade of ASIC signal transduction increases bladder capacity under normal intravesical pH conditions and alleviates bladder hyperreflexia induced by intravesical acidification and that the site responsible for this action is likely to be the dorsal root ganglia.

Ampakine pretreatment enables a single brief hypoxic episode to evoke phrenic motor facilitation

Phrenic long-term facilitation (LTF) is a sustained increase in phrenic motor output occurring after exposure to multiple (but not single) hypoxic episodes. Ampakines are a class of drugs that enhance AMPA receptor function. Ampakines can enhance expression of neuroplasticity, and the phrenic motor system is fundamentally dependent on excitatory glutamatergic currents. Accordingly, we tested the hypothesis that combining ampakine pretreatment with a single brief hypoxic exposure would result in phrenic motor facilitation lasting well beyond the period of hypoxia. Phrenic nerve output was recorded in urethane-anesthetized, ventilated, and vagotomized adult Sprague-Dawley rats. Ampakine CX717 (15 mg/kg iv; n = 8) produced a small increase in phrenic inspiratory burst amplitude and frequency, but values quickly returned to predrug baseline. When CX717 was followed 2 min later by a 5-min exposure to hypoxia ( n = 8; [Formula: see text] ~45 mmHg), a persistent increase in phrenic inspiratory burst amplitude (i.e., phrenic motor facilitation) was observed up to 60 min posthypoxia (103 ± 53% increase from baseline). In contrast, when hypoxia was preceded by vehicle injection (10% 2-hydroxypropyl-β-cyclodextrin; n = 8), inspiratory phrenic bursting was similar to baseline values at 60 min. Additional experiments with another ampakine (CX1739, 15 mg/kg) produced comparable results. We conclude that pairing low-dose ampakine treatment with a single brief hypoxic exposure can evoke sustained phrenic motor facilitation. This targeted approach for enhancing respiratory neuroplasticity may have value in the context of hypoxia-based neurorehabilitation strategies. NEW & NOTEWORTHY A single brief episode of hypoxia (e.g., 3–5 min) does not evoke long-lasting increases in respiratory motor output after the hypoxia is concluded. Ampakines are a class of drugs that enhance AMPA receptor function. We show that pairing low-dose ampakine treatment with a single brief hypoxic exposure can evoke sustained phrenic motor facilitation after the acute hypoxic episode.

Nonlinear relationship between early life stress exposure and subsequent resilience in monkeys

Retrospective correlational studies of humans suggest that moderate but not minimal or substantial early life stress exposure promotes the development of stress inoculation-induced resilience. Here we test for a nonlinear relationship between early life stress and resilience by comparing varying “doses” of early life stress. Juvenile squirrel monkeys underwent one of five treatment conditions between 17–27 weeks of age: Stress inoculation (SI) with continuous access to mother (SI + Mom; one stress element), SI without continuous access to mother (SI; two stress elements), SI without continuous access to mother and with alprazolam injection pretreatments (SI + Alz; three stress elements), SI without continuous access to mother and with vehicle injection pretreatments (SI + Veh; three stress elements), or standard housing (No SI; zero stress elements). Alprazolam was used to test whether anxiolytic medication diminished SI effects. Subjects exposed to one or two early life stressors subsequently responded with fewer indications of anxiety (e.g., decreased maternal clinging, increased object exploration, smaller cortisol increases) compared to No SI subjects. Subjects exposed to three early life stressors did not differ on most measures from one another or from No SI subjects. These findings provide empirical support for a nonlinear J-shaped relationship between early life stress exposure and subsequent resilience.

Mesenchymal Stem Cells Exhibit Both a Proinflammatory and Anti-Inflammatory Effect on Saccular Aneurysm Formation in a Rabbit Model

Several studies have demonstrated a potential interaction between mesenchymal stem cells (MSCs) and saccular aneurysms. In this study, we sought to determine whether allogenic bone marrow-derived MSCs had the ability to prevent aneurysm formation in a known rabbit elastase aneurysm model. MSCs were injected intravenously in experimental rabbits at the time of surgical creation and two weeks postcreation and compared with control rabbits receiving vehicle injection. Angiography was used to compare aneurysm measurements four weeks postcreation, and aneurysms were harvested for histological properties. Serum was collected longitudinally to evaluate cytokine alterations. Serum from control animals was also utilized to perform in vitro tests with MSCs to compare the effect of the serologic environment in animals with and without aneurysms on MSC proliferation and cytokine production. While aneurysm morphometric comparisons revealed no differences, significant cytokine alterations were observed in vitro and in vivo, suggesting both anti-inflammatory and proinflammatory processes were occurring in the presence of MSCs. Histological analyses suggested that tunica intima hyperplasia was inhibited in the presence of MSCs.

ANALISIS HASIL UJI KOMPETENSI DI LEMBAGA SERTIFIKASI PROFESI OTOMOTIF INDONESIA SE-DAERAH ISTIMEWA YOGYAKARTA TAHUN 2017

This study aims to: (1) know competence test results of competency test participants in the Indonesian Automotive Profession Certification Body from Yogyakarta Special Region (DIY) in 2017 if reviewed from the certification scheme and (2) to know the distribution of competency unit of competency test participants causing competency test participants to be declared not yet competent in LSP Otomotif Indonesia from DIY in 2017 This research is a descriptive research. The subject in this research is 198 competency test participants. Technique of collecting data is documentation. This research uses quantitative analysis technique. The result of the research shows that: (1) the competency test results of competency test participants from the Special Region of Yogyakarta in Indonesian Automotive Profession Certification Body in 2017 if reviewed based on scheme 01 (Service & Maintenance 5000 KM Motorcycles) is 70% (70) participants are declared competent and 30% (30) participants are deemed incompetent (including high category). Furthermore, in scheme 02 (Service and Maintenance 10,000 Km of Conventional Light Vehicles) 75% (15) are competent and 25% (5) declared incompetent (including category and in scheme 03 (Service and Maintenance of 10,000 Km Light Vehicle Injection System) of 61.54% (48) participants are declared competent and 38.46% (30) declared not yet competent (including very low category), (2) the distribution of competency units shows that in scheme 01, competency unit that most competent participant is OTO.SM02.001.01 (Maintaining Engine with its Components) that is equal to 73.33%, in the scheme of 02 competency units that most participants are not competent are OTO.KR02.014.01 (Maintenance / Service of Gasoline Fuel System) and OTO.KR05.011.01 (Fixing Ignition System) that is equal to 100% , whereas in the scheme of 03 competency unit that most participants are not competent is OTO.KR05.012.01 (Maintaining / Servicing and Improving Engine Management system) that is equal to 76.67%.