Environmental influences on locomotor recovery following cortical lesions in rats

Groups of rats were exposed to an enriched environment 2 hr per day for 30 days during the immediate pre- and/or postoperative period, or not at all. Animals in four of these groups sustained lesions in the bilateral sensorimotor cortex. One sham-operated control group was enriched pre- and postoperatively; a second control group was not. Animals were trained preoperatively to locomote across a narrow elevated runway. Testing on the locomotor task began 31 days after surgery and continued until preoperative performance levels were achieved. Preoperative enrichment had the most potent influence on initial deficit and recovery of locomotion. Animals that were enriched preoperatively failed to demonstrate any deficit postoperatively, and the topology of their hindlimb movement appeared to be normal. In preoperatively impoverished animals, postoperative enrichment reduced the degree of initial deficit and speeded recovery of locomotion when compared with animals not enriched at all. However, preoperatively impoverished rats demonstrated an aberrant topology of hindlimb movement even after they were "behaviorally recovered".     

Long-lasting recovery of locomotor function in chronic spinal rat following chronic combined pharmacological stimulation of serotonergic receptors with 8-OHDPAT and quipazine

In chronic spinal rats, long-term stimulation of 5-HT receptors with quipazine or 8-OHDPAT by means of daily injection, promotes robust locomotor recovery. The question of a possible potentiation between treatments when applied together was addressed. Daily injections of both 8-OHDPAT and quipazine, were performed for a month in spinal animals. Animals were placed on a treadmill and the bipedal hindlimb locomotion was tested. Motor performances (behavioural test) and locomotor parameters (EMG and kinematic) were analysed weekly during the treatment. Furthermore, the locomotor performances were evaluated during two supplemental months following the end of the treatment. Our results suggest that association of both agonists induced long-lasting positive effects on locomotor function. Motor performances were significantly better after combined injection of both drugs than when the agonists were used separately. But, the most significant and new result is that the locomotor scores did not decrease during the weeks that followed the end of the treatment. These results suggests a long-lasting and 5-HT-dependent reorganisation of spinal networks.     

Delayed inhibition of Nogo-A does not alter injury-induced axonal sprouting but enhances recovery of cognitive function following experimental traumatic brain injury in rats

Traumatic brain injury causes long-term neurological motor and cognitive deficits, often with limited recovery. The inability of CNS axons to regenerate following traumatic brain injury may be due, in part, to inhibitory molecules associated with myelin. One of these myelin-associated proteins, Nogo-A, inhibits neurite outgrowth in vitro, and inhibition of Nogo-A in vivo enhances axonal outgrowth and sprouting and improves outcome following experimental CNS insults. However, the involvement of Nogo-A in the neurobehavioral deficits observed in experimental traumatic brain injury remains unknown and was evaluated in the present study using the 11C7 monoclonal antibody against Nogo-A. Anesthetized, male Sprague-Dawley rats were subjected to either lateral fluid percussion brain injury of moderate severity (2.5-2.6 atm) or sham injury. Beginning 24 h post-injury, monoclonal antibody 11C7 (n=17 injured, n=6 shams included) or control Ab (IgG) (n=16 injured, n=5 shams included) was infused at a rate of 5 microl/h over 14 days into the ipsilateral ventricle using osmotic minipumps connected to an implanted cannula. Rats were assessed up to 4 weeks post-injury using tests for neurological motor function (composite neuroscore, and sensorimotor test of adhesive paper removal) and, at 4 weeks, cognition was assessed using the Morris water maze. Hippocampal CA3 pyramidal neuron damage and corticospinal tract sprouting, using an anterograde tracer (biotinylated dextran amine), were also evaluated. Brain injury significantly increased sprouting from the uninjured corticospinal tract but treatment with monoclonal antibody 11C7 did not further increase the extent of sprouting nor did it alter the extent of CA3 cell damage. Animals treated with 11C7 showed no improvement in neurologic motor deficits but did show significantly improved cognitive function at 4 weeks post-injury when compared with brain-injured, IgG-treated animals. To our knowledge, the present findings are the first to suggest that (1) traumatic brain injury induces axonal sprouting in the corticospinal tract and this sprouting may be independent of myelin-associated inhibitory factors and (2) that post-traumatic inhibition of Nogo-A may promote cognitive recovery unrelated to sprouting in the corticospinal tract or neuroprotective effects on hippocampal cell loss following experimental traumatic brain injury.     

Human mesenchymal stem cell transplantation promotes functional recovery following acute spinal cord injury in rats

Many attempts have been made in animals to produce cellular regeneration in the spinal cord using a variety of transplanted cell types. The present study was to investigate whether transplantation of human mesenchymal stem cells (hMSCs) into the spinal cord after contusion injury promotes a functional outcome. Spinal cord injury (SCI) was induced using an NYU impactor and hMSCs were transplanted 1 week after SCI. Behavioral testing was performed weekly for 2 months. Somatosensory (SSEPs) and motor evoked potentials (MEPs) were recorded to determine functional recovery. Hindlimb performance was modestly improved in the transplanted group based on BBB scaling and pain tests. SSEP latencies in the transplanted group were significantly shorter than in the media-treated group. Pathologically, LacZ and hTau positive cells were located at the injury and adjacent sites. The data indicate improvement in functional outcome in animals treated with hMSC transplantation compared to media-treated animals.     

Zinc has no effect on IL-3-mediated apoptosis of BAF-3 cells but enhances CD95-mediated apoptosis of jurkat cells

The feasibility of using a zinc-inducible gene expression system for the study of apoptosis-controlling genes in BAF-3 murine B cells and Jurkat human T cells was evaluated. Initially, cell sensitivity to a range of zinc concentrations was examined. It was found that zinc concentrations above 60 microM were toxic to BAF-3 cells and those above 50 microM were toxic to Jurkat cells. Secondly, the zinc concentration required to achieve maximal gene expression was examined. BAF-3 cells transiently transfected with the pMTCB6+/luciferase vector were exposed to zinc concentrations ranging from 0-120 microM, whilst stably transfected Jurkat cells were exposed to 0-70 microM zinc. At zinc concentrations nontoxic to each cell type, the maximum induction achieved was 20-fold (at 60 microM) in BAF-3 cells, and 7.5-fold (at 50 microM) in Jurkat cells. Thirdly, the effect of zinc on apoptosis was examined. It was shown that exposure to nontoxic zinc concentrations had no effect on IL-3 withdrawal-mediated apoptosis of BAF-3 cells. However, in the case of Jurkat cells, pre-exposure to zinc augmented CD95-mediated apoptosis. These results illustrate the importance of characterizing individual cell lines when using zinc-inducible gene expression systems.     

Route of nutrition has no effect on the development of infectious complications

BACKGROUND: Infection is a serious complication of nutritional support, causing a high rate of mortality and morbidity. Critically ill patients having nutritional support are prone to infectious complications. Questions regarding the effects of the route of nutrition in infectious complications have been asked. We aimed to determine the relationship between the route of nutrition and the risk of developing infectious complications in severely ill patients on nutritional support in an intensive care unit. METHODS: A retrospective review was performed on the files of 144 severely ill patients who had either enteral or parenteral nutrition during follow-up in an intensive care unit. The primary diagnoses of patients were heterogenous. RESULTS: Sixty-eight (35.8%) of them acquired novel infections during the hospitalization period. Forty-nine and 19 of the 68 infected patients had enteral and parenteral nutrition support, respectively. Seventy-six (40%) of the patients were free of infection. Fifty-one of 76 infection-free patients had enteral nutrition support, and 25 of them had parenteral nutrition support. Pulmonary infections, urinary tract infections, catheter infections and septicemia were the most frequent types of infectious complications. There was no significant difference in the rate of infectious complications between enteral nutrition and parenteral nutrition groups (p > 0.05). CONCLUSION: We conclude that the route of the nutritional support in severely ill patients having nutritional support in an intensive care unit does not affect the rate of infectious complications. We think that comorbid medical conditions and the need of intensive care unit support are more important parameters that determine the risk of development of infectious complications.     

慢性压迫性神经损伤大鼠中透明质酸合成变化及其在神经源性疼痛中的作用

慢性神经源性疼痛由于其发病机制尚不完全明确,目前还没有十分有效的治疗手段;神经损伤后炎症反应和免疫调节机制在疼痛的发生中发挥着重要作用,透明质酸(HA)近来被认为是炎症和免疫调节中一个重要的调节分子。为了进一步研究HA是否参与到神经损伤后的病理过程中,我们检测了慢性压迫性神经损伤(CC I)大鼠损伤神经的HA含量。结果显示:与正常神经比较,HA的含量在损伤后7 d明显增加,HA合成酶(HAS)的表达也明显上调。4-甲基伞形酮(4-MU)是HAS的一种抑制剂,我们通过给予4-MU抑制HA的合成,研究HA在慢性神经源性疼痛中的作用,发现给药组CC I大鼠损伤足对热痛刺激的敏感性低于未给药组,同时IL-1β的表达量低于未给药组。以上结果提示HA可能通过对炎症因子的调控参与到损伤后的疼痛机制中,这一结果将有助于慢性神经源性疼痛的治疗。     

Pharmacological manipulation with the descending serotonergic system or transection of the mouse spinal cord has no effect on ependymal ultrastructure

In order to investigate an effect of descending nerve fibres on mouse spinal cord ependymal ultrastructure, pharmacological manipulation with the serotonergic system or transection of the spinal cord was done. Biochemical analysis showed an 83% reduction of serotonin content in spinal cord tissue after p-chlorophenylalanine injections and a 93% reduction after transection. However, none of the experimental animals showed changes in ependymal ultrastructure compared to control animals as revealed by electron microscopy.     

Phenolic glycolipid 1 of Mycobacterium leprae causes nonspecific inflammation but has no effect on cell-mediated responses in mice

The involvement of the phenolic glycolipid from Mycobacterium leprae in cell-mediated immunity has been investigated in this study. The phenolic glycolipid itself does not appear to stimulate cell-mediated immunity directly, as shown by its failure to elicit a classical delayed-type hypersensitivity response in mice immunized with M. leprae or to stimulate M. leprae-immune lymph node cells in a lymphoproliferative assay. Intradermal vaccination with the phenolic glycolipid failed to influence the growth of M. leprae in mouse footpads. A nonspecific inflammatory response to the sonicated glycolipid was observed in mice vaccinated with whole M. leprae and in control animals. No evidence was obtained for any adjuvant or suppressive effect on cell-mediated immunity by the phenolic glycolipid either to M. leprae or an unrelated antigen (sheep erythrocytes); neither sensitization nor elicitation to either antigen was affected.     

The different mechanisms of peripheral and central TLR4 on chronic postsurgical pain in rats

Chronic postsurgical pain (CPSP) is a common complication after surgery; however, the underlying mechanisms of CPSP are poorly understood. As one of the most important inflammatory pathways, the Toll-like receptor 4/nuclear factor-kappa B (TLR4/NF-κB) signaling pathway plays an important role in chronic pain. However, the precise role of the TLR4/NF-κB signaling pathway in CPSP remains unclear. In the present study, we established a rat model of CPSP induced by skin/muscle incision and retraction (SMIR) and verified the effects and mechanisms of central and peripheral TLR4 and NF-κB on hyperalgesia in SMIR rats. The results showed that TLR4 expression was increased in both the spinal dorsal horn and dorsal root ganglia (DRGs) of SMIR rats. However, the TLR4 expression pattern in the spinal cord was different from that in DRGs. In the spinal cord, TLR4 was expressed in both neurons and microglia, whereas it was expressed in neurons but not in satellite glial cells in DRGs. Further results demonstrate that the central and peripheral TLR4/NF-κB signaling pathway is involved in the SMIR-induced CPSP by different mechanisms. In the peripheral nervous system, we revealed that the TLR4/NF-κB signaling pathway induced upregulation of voltage-gated sodium channel 1.7 (Nav1.7) in DRGs, triggering peripheral hyperalgesia in SMIR-induced CPSP. In the central nervous system, the TLR4/NF-κB signaling pathway participated in SMIR-induced CPSP by activating microglia in the spinal cord. Ultimately, our findings demonstrated that activation of the peripheral and central TLR4/NF-κB signaling pathway involved in the development of SMIR-induced CPSP.     

Almitrine has no effect on gas exchange after bilateral carotid body resection in severe chronic airflow obstruction

Using a double-blind cross-over design, a single dose of 100 mg almitrine bismesylate and placebo were administered orally to eight patients with chronic airflow obstruction having undergone bilateral carotid body resection (BCBR) up to two years earlier to alleviate their extreme dyspnoea. In an open study, two other patients have been given almitrine before and three weeks after BCBR. In all patients, arterial blood gases, ventilation and breathing patterns, neuromuscular drive and hypoxic responsiveness have been studied before and three hours after drug administration. Almitrine failed to improve gas exchange in the patients with BCBR, nor did it affect ventilation, ventilatory or hypoxic drive. In the patients studied before and after BCBR, almitrine only improved gas exchange significantly before BCBR. It is concluded that in man almitrine acts solely as a peripheral chemoreceptor agonist and that the well-documented improvement in V/Q relationship is mediated through carotid body stimulation.     

Recovery of locomotor activity in the adult chronic spinal rat after sublesional transplantation of embryonic nervous cells: specific role of serotonergic neurons

 Locomotor movements are programmed in a specialised neuronal network that is localised in the central nervous system and referred to as the central pattern generator (CPG) for locomotion. This CPG can be activated by pharmacological agents such as monoamines. The aim of the present study was to try to activate the CPGs by using cells that are supposed to release serotonin locally. Adult chronic spinal rats were injected with embryonic brainstem neurons within the spinal cord under a thoracic transection. This procedure resulted in a monoaminergic reinnervation of the lumbar enlargement. With the help of a specific neurotoxin for noradrenergic neurons (6-hydroxydopamine), it was possible to isolate the serotonergic system. After such transplantation of monoaminergic neurons and even with serotonergic neurons alone, a bilateral, alternating, rhythmic locomotor-like activity recovered in hindlimbs. Furthermore, this locomotor-like activity was clearly facilitated when the re-uptake of serotonin was blocked by zimelidine. Therefore, we conclude that transplanted embryonic serotonergic neurons are able to activate the CPG for locomotion. Received: 5 January 1996 / Accepted: 20 August 1996     

Alkylation of striatal dopamine receptors abolishes stereotyped behavior but has no effect on dopamine stimulated adenylate cyclase activity

The role of dopamine stimulated adenylate cyclase (AC) activity in behaviours elicited by apomorphine stimulation of striatal dopamine receptors was investigated. Rats were treated with the irreversible dopamine receptor alkylating agent N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ) alone or after pretreatment with either a D1 or D2 receptor antagonist and subsequently challenged with apomorphine. Animals that received no antagonist pretreatment showed significantly decreased striatal concentrations of D1 and D2 receptors and an abolition of apomorphine induced sniffing behaviour despite showing no change in striatal AC activity. In the groups which received antagonist pretreatment the reduction in the sniffing response paralleled the reduction in D2 receptor concentration whereas the incidence of vacuous oral movements was inversely related. In no case were the behavioural responses associated with changes in AC activity. We conclude that these behavioural effects observed in response to dopamine stimulation by apomorphine may be mediated through another second messenger system.     

Induced Syk deletion leads to suppressed allergic responses but has no effect on neutrophil or monocyte migration in vivo

The spleen tyrosine kinase (Syk) is a key mediator of immunoreceptor signaling in immune cells. Thus, interfering with the function of Syk by genetic deletion or pharmacological inhibition might influence a variety of allergic and autoimmune processes. Since conventional Syk knockout mice are not viable, studies addressing the effect of Syk deletion in adult animals have been limited. To further explore functions of Syk in animal models of allergy and to shed light on the role of Syk in the in vivo migration of neutrophils and monocytes, we generated inducible Syk knockout mice. These mice harbor a floxed Syk gene and a tamoxifen-inducible Cre recombinase under the control of the ubiquitously active Rosa26-promoter. Thus, treatment of mice with tamoxifen leads to the deletion of Syk in all organs. Syk-deleted mice were analyzed in mast cell-dependent models and in models focusing on neutrophil and monocyte migration. We show that Syk deletion in adult mice reduces inflammatory responses in mast cell-driven animal models of allergy and asthma but has no effect on the migration of neutrophils and monocytes. Therefore, the inducible Syk knockout mice presented here provide a valuable tool to further explore the role of Syk in disease-related animal models.     

Transient cold pain has no effect on cutaneous vasodilatation induced by capsaicin: a randomized-control-crossover study in healthy subjects

Cooling the skin induces sympathetically driven vasoconstriction, along with some vasoparalytic dilatation at lowermost temperatures. Neurogenic inflammation, on the other hand, entails vasodilatation. In the present study, we examined the dynamic vasomotor balance of capsaicin-induced vasodilatation within the area of the induced neurogenic inflammation, with and without superimposed cooling. In a randomized-control-crossover fashion, a sample of 14 healthy volunteers participated in three experiments: (1) exposure to each 0°C cold pain stimulus and a neutral 30°C stimulus (control) for 30 s to the volar forearms by contact thermal thermode (1.6×1.6 cm²), (2) injection of 50 μg intradermal capsaicin without cooling and (3) injection of capsaicin followed by application of 0°C cold pain stimulation for 30 s within the area of the secondary hyperalgesia. Repetitive vascular measurements over skin area of 4.0×4.0 cm² of blood flux (BF) were acquired before and during the 5 min after stimulation. A marked increase in BF (i.e. vasodilatation) at the location of the cold stimulus in comparison to control (30°C) (F=11.97, p=0.004) within the first 3 min was demonstrated. Two-way repeated-measures ANOVA indicated no interaction between the experimental conditions (capsaicin with or without cold) and time (F=0.934, p=0.454). The cold pain stimulation was found to be insignificant in its influence on BF evoked by capsaicin (F=0.018, p=0.894). The results of our study indicate that (1) transient cooling causes significant vasodilatation, (2) intradermal injection of capsaicin is dominant in inducing vasodilatation, and (3) the cold-pain-evoked vasodilatation has no modulative effect on the capsaicin-evoked cutaneous vasodilatation.     

Longitudinal myelotomy of lumbar spinal cord has little effect on coordinated locomotor activities of bilateral hindlimbs of the chronic cats

Phase relations between muscle activities of bilateral hindlimbs during walking were investigated in chronic cats whose spinal cord was longitudinally separated into halves in order to investigate if segmental commissural connections are essential for the coordination. EMGs were recorded from bilateral triceps brachii, bilateral vastus lateralis and one tibialis anterior muscles during overground locomotion. The results show that coordination between bilateral hindlimbs is well preserved after the longitudinal myelotomy of the lumbar cord, indicating structures located supra-lumbar cord are essentially important for the coordination.     

Correction of endothelial dysfunction after selective homocysteine lowering gene therapy reduces arterial thrombogenicity but has no effect on atherogenesis

Hyperhomocysteinemia is an independent risk factor for ischemic cardiovascular diseases, but its causal role in atherothrombosis remains controversial. Proatherogenic and/or prothrombotic effects may underlie the potential causal relation between hyperhomocysteinemia and cardiovascular events. Here, the effects of selective lowering of plasma homocysteine, plasma cholesterol, or both on endothelial function and on atherogenesis in male hyperlipidemic and hyperhomocysteinemic C57BL/6 low-density lipoprotein receptor (LDLr)^−/−/cystathionine-β-synthase (CBS)^+/−-deficient mice were investigated. Second, we evaluated whether selective homocysteine lowering has anti-thrombotic effects in a model of arterial thrombosis. A hyperhomocysteinemic and atherogenic diet was started at the age of 12 weeks. Three weeks later, gene transfer was performed with E1E3E4-deleted adenoviral vectors for hepatocyte-restricted overexpression of CBS (AdCBS) or of the LDLr (AdLDLr), or with the control vector Adnull. In a fourth group, AdCBS and AdLDLr were co-administered. Selective homocysteine lowering but not selective cholesterol lowering restored endothelial function at 6 weeks after gene transfer. Intimal area in the aortic root and in the brachiocephalic artery at 13 weeks was more than 100-fold (p < 0.001) smaller in AdLDLr and AdCBS/AdLDLr mice than in control mice and AdCBS mice. No differences in intimal area were observed between control mice and AdCBS mice. In a model of carotid artery thrombosis, the average time to first occlusion and to stable occlusion were 1.9-fold (p < 0.01) and 2.1-fold longer (p < 0.01), respectively, in AdCBS-treated mice than in control mice. Taken together, these data show that correction of endothelial dysfunction following selective homocysteine lowering has anti-thrombotic but no anti-atherogenic effects.     

内质网应激在慢性间歇低氧幼鼠脑损害中的作用

目的:探讨内质网应激在慢性间歇低氧幼鼠脑损害中的作用机制及salubrinal的干预作用。方法:取SPF级健康雄性SD幼鼠64只,随机分为8组:间歇低氧(intermittent hypoxia,IH)2、4周组(2IH、4IH),对照(control,C)2、4周组(2C、4C),Salubrinal(SAL)干预2、4周组(2SAL、4SAL),二甲基亚砜(DMSO)溶剂对照2、4周组(2DMSO、4DMSO),每组8只。八臂迷宫测试各组幼鼠参考记忆错误(RME)、工作记忆错误(WME)及总错误(TE)次数,观察海马神经元凋亡变化,测定超氧化物歧化酶(SOD)活性,及内质网应激标志物C/EBP同源蛋白(CHOP)、磷酸化真核翻译起始因子2α(p-e IF2α)和磷酸化蛋白激酶R样内质网激酶(p-PERK)的蛋白水平。结果:与相应对照2C、4C组比较,间歇低氧2IH、4IH组幼鼠的RME、WME和TE升高(P0.01),海马神经元凋亡指数(AI)升高(P0.01),SOD活性下降(P0.01),p-PERK和CHOP蛋白水平升高(P0.01),p-e IF2α蛋白水平下降(P0.05),4周组最明显;与对应间歇性低氧2IH、4IH组比较,药物干预组2SAL、4SAL组RME、WME和TE次数下降(P0.05),AI下降(P0.01),SOD活性升高(P0.01),p-e IF2α的蛋白水平升高(P0.01),CHOP表达下降(P0.01)。结论:慢性间歇低氧可上调记忆相关脑区p-PERK表达,启动内质网应激,从而诱导CHOP所介导的细胞凋亡,可能在慢性间歇低氧所致脑损伤中起重要作用。Salubrinal选择性抑制e IF2α去磷酸化,下调CHOP蛋白的水平,提高SOD活性,从而缓解内质网应激,减轻氧化应激,减少细胞凋亡。     

Lactobacillus rhamnosus reduces parasite load on Toxocara canis experimental infection in mice,but has no effect on the parasite in vitro

Parasitology Research - Human toxocariasis is a neglected global parasitic zoonosis. The efficacy of drug treatment for this disease has been hindered by the biological complexity of the main...