M. Vaneker

α7 Nicotinic acetylcholine receptor agonist GTS-21 attenuates ventilator-induced tumour necrosis factor-α production and lung injury

BACKGROUNDnMechanical ventilation (MV) induces an inflammatory response that can lead to lung injury. The vagus nerve can limit the inflammatory response through the cholinergic anti-inflammatory pathway. We evaluated the effects of stimulation of the cholinergic anti-inflammatory pathway with the selective partial α7 nicotinic acetylcholine receptor (α7nAChR) agonist GTS-21 on inflammation and lung injury induced by MV using clinically relevant ventilator settings. Furthermore, we investigated whether altering endogenous cholinergic signalling, by administration of the non-specific nAChR antagonist mecamylamine and the peripherally acting acetylcholinesterase inhibitor neostigmine, modulates the MV-induced inflammatory response.nnnMETHODSnC57BL6 mice were injected i.p. with either the selective α7nAChR agonist GTS-21 (8 mg kg(-1)), the acetylcholinesterase inhibitor neostigmine (80 μg kg(-1)), the nAChR antagonist mecamylamine (1 mg kg(-1)), or a placebo; followed by 4 h of MV (8 ml kg(-1), 1.5 cm H(2)O PEEP).nnnRESULTSnMV resulted in release of cytokines in plasma and lungs compared with unventilated mice. Lung and plasma levels of tumour necrosis factor (TNF)-α, but not of interleukin-10, were lower in GTS-21-treated animals compared with placebo (P<0.05). In addition, GTS-21 lowered the alveolar-arterial gradient, indicating improved lung function (P=0.04). Neither neostigmine nor mecamylamine had an effect on MV-induced inflammation or lung function.nnnCONCLUSIONSnMV with clinically relevant ventilator settings results in pulmonary and systemic inflammation. Stimulation of the cholinergic anti-inflammatory pathway with GTS-21 attenuates MV-induced release of TNF-α, which was associated with reduced lung injury. Modulation of endogenous cholinergic signalling did not affect the MV-induced inflammatory response. Selective stimulation of the cholinergic anti-inflammatory pathway may represent new treatment options for MV-induced lung injury.

Patients initially diagnosed as 'warm' or 'cold' CRPS 1 show differences in central sensory processing some eight years after diagnosis: a quantitative sensory testing study.

We used quantitative sensory testing (QST) to gain further insight into mechanisms underlying pain in CRPS 1. Specific goals were: (1) to identify altered patterns of sensory processing some 8 years after diagnosis, (2) to document differences in sensory processing between ‘warm’ and ‘cold’ diagnostic subgroups, (3) to determine relationships between changed sensory processing and disease progression regarding pain. The study was performed on a cohort of patients (n=47) clinically diagnosed with CRPS 1 of one upper extremity approximately 8 years previously. Pain was quantified by VAS and MacGill Pain Questionnaire (MPQ), and all subjects underwent electrical and mechanical QST. Cold patients (n=13) had poorer MPQ scores than warm ones (n=34), and more pain on electrical stimulation. Their evoked pain increased with disease progression and correlated with clinical pain measures. For both diagnostic subgroups, thresholds to pressure pain were lower on the affected extremity and with disease progression. Eight years after original diagnosis, cold CRPS 1 patients have poorer clinical pain outcomes and show persistent signs of central sensitisation correlating with disease progression. The latter is not the case for warm CRPS 1 patients. Both diagnostic subgroups show greater pressure hyperalgesia on the affected limb and with disease progression. QST may prove useful in the subdiagnosis of CRPS 1 and in quantifying its progression, with both applications warranting further investigation for clinical and research use.

Effects of Vagus Nerve Stimulation and Vagotomy on Systemic and Pulmonary Inflammation in a Two-Hit Model in Rats

Pulmonary inflammation contributes to ventilator-induced lung injury. Sepsis-induced pulmonary inflammation (first hit) may be potentiated by mechanical ventilation (MV, second hit). Electrical stimulation of the vagus nerve has been shown to attenuate inflammation in various animal models through the cholinergic anti-inflammatory pathway. We determined the effects of vagotomy (VGX) and vagus nerve stimulation (VNS) on systemic and pulmonary inflammation in a two-hit model. Male Sprague-Dawley rats were i.v. administered lipopolysaccharide (LPS) and subsequently underwent VGX, VNS or a sham operation. 1 hour following LPS, MV with low (8 mL/kg) or moderate (15 mL/kg) tidal volumes was initiated, or animals were left breathing spontaneously (SP). After 4 hours of MV or SP, rats were sacrificed. Cytokine and blood gas analysis was performed. MV with 15, but not 8 mL/kg, potentiated the LPS-induced pulmonary pro-inflammatory cytokine response (TNF-α, IL-6, KC: p<0.05 compared to LPS-SP), but did not affect systemic inflammation or impair oxygenation. VGX enhanced the LPS-induced pulmonary, but not systemic pro-inflammatory cytokine response in spontaneously breathing, but not in MV animals (TNF-α, IL-6, KC: p<0.05 compared to SHAM), and resulted in decreased pO2 (p<0.05 compared to sham-operated animals). VNS did not affect any of the studied parameters in both SP and MV animals. In conclusion, MV with moderate tidal volumes potentiates the pulmonary inflammatory response elicited by systemic LPS administration. No beneficial effects of vagus nerve stimulation performed following LPS administration were found. These results questions the clinical applicability of stimulation of the cholinergic anti-inflammatory pathway in systemically inflamed patients admitted to the ICU where MV is initiated.

Isoflurane attenuates pulmonary interleukin‐1β and systemic tumor necrosis factor‐α following mechanical ventilation in healthy mice

Background: Mechanical ventilation (MV) induces an inflammatory response in healthy lungs. The resulting pro‐inflammatory state is a risk factor for ventilator‐induced lung injury and peripheral organ dysfunction. Isoflurane is known to have protective immunological effects on different organ systems. We tested the hypothesis that the MV‐induced inflammatory response in healthy lungs is reduced by isoflurane.

Behavior of neuropathic pain in mice following chronic constriction injury comparing silk and catgut ligatures

IntroductionNeuropathic pain is defined as pain arising as a direct consequence of a lesion or disease affecting the somatosensory system and is common after surgery. Neuropathic pain can persist without an obvious injury. In this study we aim to validate a murine chronic constriction injury model as a model for neuropathic pain research and determine if silk or catgut ligatures induced most stable neuropathic pain behavior.MethodsIn this study mice underwent chronic constriction or sham surgery. Mice were tested on cutaneous hyperalgesia with the cumulative reaction time in the acetone test, on allodynia with the cumulative reaction time and number of lifts in the cold plate test and the maximal force before withdrawal in von Frey test.ResultsIn the acetone test neuropathic pain was seen in CCI mice, but not in sham mice. Hyperalgesia was present postoperatively in CCI mice compared with preoperatively. In the cold plate test cumulative reaction time and number of lifts were higher in the ipsilateral hind paw than in the contralateral hind paw and sham mice. Postoperative measurements were higher than preoperatively. In the von Frey test the postoperative measurements were lower in the ipsilateral hind paw than preoperatively, while the contralateral hind paw showed an increase in maximal force before withdrawal. The contralateral hind paw showed more difference with sham mice than the ipsilateral hind paw. Silk ligatures showed more stable neuropathic pain behavior. In the acetone test, the cold plate test and the von Frey test the mice scored higher on neuropathic pain having silk ligatures, compared with catgut ligatures.ConclusionIn this study we validated a murine CCI model for neuropathic pain behavior. In the murine CCI model it appears that silk ligatures demonstrate more stable neuropathic pain behaviors than catgut ligatures in de CCI model.

Mechanical ventilation of mice under general anesthesia during experimental procedures

Mechanical ventilation is frequently used in patients under general anesthesia during invasive procedures. Invasive animal experiments similarly require the maintenance of normal hemodynamic and pulmonary parameters during long-term general anesthesia. The authors describe a method for mechanical ventilation of mice. Mice were ventilated and monitored for up to 8 h of general anesthesia during surgery. Hemodynamic and pulmonary parameters remained within the normal ranges. The authors believe that this ventilation technique can be of great value for experimental procedures in mice that require general anesthesia.

The role of poly (ADP-ribose) polymerase in ventilator-induced lung injury

With interest we have read the paper by Kim and colleagues reporting the role of poly (ADP-ribose) polymerase (PARP) in ventilator-induced lung injury (VILI) in healthy mice [1]. n nSome issues have not been addressed appropriately. The authors show increased levels of TNFα in lung homogenate after 2 hours of lung-protective ventilation (LPV). Previous data from our laboratory have shown in the healthy mouse lung that so-called protective mechanical ventilation (tidal volume, 8 ml/kg; peak airway pressure, 10 to 12 cmH2O; positive end-expiratory pressure, 4 cmH2O) induces a pulmonary inflammatory response [2]. In addition to elevated levels of TNFα, we found increased expression of IL-1β, IL-6, and keratinocyte-derived chemokine in the lung homogenate and found an increased number of pulmonary leucocytes in mice mechanically ventilated for 2 hours. Electron microscopy revealed evidence for type I pneumocyte membrane disruption and endothelial detachment, indicating structural injury. In line with the findings by Kim and colleagues, the wet/dry ratio was not affected after 2 hours of mechanical ventilation – although in our study 4 hours of protective ventilation did increase the wet/dry ratio. n nThe so-called LPV-induced pulmonary inflammation can therefore occur without activation of PARP. It is possible that initiation of inflammation precedes the activation of PARP. Do the authors have any information on the time dependency of PARP activation in relation to the activation of proinflammatory cytokines? In addition, it would be of interest to identify the type of cells exhibiting elevated PARP activity – for instance, perhaps by double staining with leukocyte markers. In our opinion, the clinical relevance of the study is limited due to very high peak airway pressures used.