Effect of acute isooxic hypercapnia on oxidative activity of systemic neutrophils in endotoxemic rabbits.

INTRODUCTION: Whether carbon dioxide (CO2) affects systemic oxidative phenomena under conditions of endotoxemia is not sufficiently clarified. The study aimed to assess the impact of moderate acute hypercapnia on the respiratory burst of circulating neutrophils in mechanically ventilated endotoxemic rabbits. MATERIAL AND METHODS: Twenty-four endotoxemic rabbits were mechanically ventilated with standard or CO2-enriched gas mixture in order to obtain isooxic hypercapnia. At a baseline point and following 180 min of hypercapnic ventilation, luminol-dependent chemiluminescence (CL) of circulating neutrophils and serum 2-thiobarbituric acid reactive substance (TBARS) concentrations were measured. Throughout the study, leukocyte and neutrophil counts, pH status, circulatory parameters and body temperature were also assessed. RESULTS: Following 180 min of hypercapnic ventilation, opsonized zymosan (OZ)-stimulated neutrophils showed lower CL vs. the control group (p = 0.004). Other parameters studied were not affected. CONCLUSIONS: Short-term isooxic hypercapnia in endotoxemic rabbits preserves circulating neutrophil count pattern and reactive oxygen species (ROS) generation, but it may reduce phagocytosis.

Learning-related feedforward inhibitory connectivity growth required for memory precision.

In the adult brain, new synapses are formed and pre-existing ones are lost, but the function of this structural plasticity has remained unclear. Learning of new skills is correlated with formation of new synapses. These may directly encode new memories, but they may also have more general roles in memory encoding and retrieval processes. Here we investigated how mossy fibre terminal complexes at the entry of hippocampal and cerebellar circuits rearrange upon learning in mice, and what is the functional role of the rearrangements. We show that one-trial and incremental learning lead to robust, circuit-specific, long-lasting and reversible increases in the numbers of filopodial synapses onto fast-spiking interneurons that trigger feedforward inhibition. The increase in feedforward inhibition connectivity involved a majority of the presynaptic terminals, restricted the numbers of c-Fos-expressing postsynaptic neurons at memory retrieval, and correlated temporally with the quality of the memory. We then show that for contextual fear conditioning and Morris water maze learning, increased feedforward inhibition connectivity by hippocampal mossy fibres has a critical role for the precision of the memory and the learned behaviour. In the absence of mossy fibre long-term potentiation in Rab3a(-/-) mice, c-Fos ensemble reorganization and feedforward inhibition growth were both absent in CA3 upon learning, and the memory was imprecise. By contrast, in the absence of adducin 2 (Add2; also known as ß-adducin) c-Fos reorganization was normal, but feedforward inhibition growth was abolished. In parallel, c-Fos ensembles in CA3 were greatly enlarged, and the memory was imprecise. Feedforward inhibition growth and memory precision were both rescued by re-expression of Add2 specifically in hippocampal mossy fibres. These results establish a causal relationship between learning-related increases in the numbers of defined synapses and the precision of learning and memory in the adult. The results further relate plasticity and feedforward inhibition growth at hippocampal mossy fibres to the precision of hippocampus-dependent memories.

Effects of local anesthetics on the respiratory burst of cord blood neutrophils in vitro.

BACKGROUND: Whether local anesthetics exert anti-inflammatory effects in fetal and newborn systemic neutrophils is unclear. The aim of the present study was to assess the effects of bupivacaine and lidocaine on the respiratory burst of cord blood neutrophils in vitro compared with adult cells. METHODS: Whole cord blood (n = 12) and control adult blood samples (n = 7) were incubated with bupivacaine (0.0005, 0.005, 0.05, 1 mmol/l) and lidocaine (0.002, 0.02, 0.2, 4 mmol/l) for 1 and 4 h. The production of reactive oxygen species (ROS) by unstimulated neutrophils and the phorbol myristate acetate-induced oxidative burst were assessed by flow cytometry. A subset of neutrophils showing high fluorescence intensity (rho+) was analyzed separately. RESULTS: After 1 h incubation, local anesthetics decreased the respiratory burst in whole cord blood and adult neutrophils in a similar manner. At the clinically relevant concentration of 0.0005 mmol/l, bupivacaine was active, but its effect in cord blood cells could not be detected after 4 h. The cord blood rho+ cell subset was unresponsive to the inhibitory action of bupivacaine. In rho+ neutrophils, basal ROS production was stimulated by lidocaine at the lowest concentration tested. CONCLUSION: Bupivacaine and lidocaine can decrease the respiratory burst in neutrophils of term newborns.

Mouse δ opioid receptors are located on presynaptic afferents to hippocampal pyramidal cells.

Delta opioid receptors participate in the control of chronic pain and emotional responses. Recent data have also identified their implication in drug-context associations pointing to a modulatory role on hippocampal activity. We used fluorescent knock-in mice that express a functional delta opioid receptor fused at its carboxy terminus with the green fluorescent protein in place of the native receptor to investigate the receptor neuroanatomical distribution in this structure. Fine mapping of the pyramidal layer was performed in hippocampal acute brain slices and organotypic cultures using fluorescence confocal imaging, co-localization with pre- and postsynaptic markers and correlative light-electron microscopy. The different approaches concurred to identify delta opioid receptors on presynaptic afferents to glutamatergic principal cells. In the latter, only scarce receptors were detected that were confined within the Golgi or vesicular intracellular compartments with no receptor present at the cell surface. In the mouse hippocampus, expression of functional delta opioid receptors is therefore mostly associated with interneurons emphasizing a presynaptic modulatory effect on the pyramidal cell firing rate.

The influence of bupivacaine on the whole blood chemiluminescence in laboring women--a preliminary report.

OBJECTIVE: Local anesthetics are able to inhibit inflammatory phenomena. The influence of bupivacaine broadly applied in obstetrical anesthesia on the reactive oxygen species (ROS) production is a matter of controversy. The study was aimed at elucidation of the influence of racemic bupivacaine on the opsonized zymosan (OZ) stimulated-peripheral blood chemiluminescence (CL) in laboring women, reflecting the reactive oxygen species (ROS) production associated with phagocytosis. MATERIAL AND METHODS: Blood samples drawn from 8 healthy parturients in active spontaneous labor and from 5 healthy non-pregnant controls were incubated with the 0.3, 30, and 3000 microM bupivacaine concentrations and then luminol-dependent OZ-stimulated whole blood CL was assessed. RESULTS: Bupivacaine depressed CL; however the inhibitory effect was significant only at the highest, clinically irrelevant concentration (3000 microM), in parturients being comparable to that observed in non-pregnant women. CONCLUSIONS: Bupivacaine at clinically relevant concentrations does not influence ROS production accompanying phagocytosis in peripheral blood of laboring women. The effect is comparable in parturients and non-pregnant controls.

The influence of the time of day on midazolam pharmacokinetics and pharmacodynamics in rabbits.

BACKGROUND: This study evaluates the time-of-day effect on midazolam and 1-OH midazolam pharmacokinetics, and on the sedative pharmacodynamic response in rabbits. Also, circadian fluctuations in rabbits' vital signs, such as the blood pressure, heart rate and body temperature were examined. The water intake was measured in order to confirm the presence of the animals' diurnal activity. The secondary aim involved the comparison of two methods of data analysis: a noncompartmental and a population modeling approach. METHODS: Twelve rabbits were sedated with intravenous midazolam 0.35 mg/kg at four local times: 09.00, 14.00, 18.00 and 22.00 h. Each rabbit served as its own control by being given a single infusion at the four different times of the day on four separate occasions. The values of the monitored physiological parameters were recorded during the experiment and arterial blood samples were collected for midazolam assay. The pedal withdrawal reflex was used as the measurement of the sedation response. Two and one compartmental models were successfully used to describe midazolam and 1-OH midazolam pharmacokinetics. The categorical pharmacodynamic data were described with a logistic model. RESULTS AND CONCLUSIONS: We did not find any time-of-day effects for the pharmacokinetic and pharmacodynamics parameters of midazolam. For 1-OH midazolam, statistically significant time-of-day differences in the apparent volume of distribution and clearance were noticed. They corresponded well with the rabbits' water intake. The noncompartmental and model-based parameters were essentially similar. However, more information can be obtained from the population model and this method should be preferred in chronopharmacokinetic and chronopharmacodynamic studies.

ß-Adducin is required for stable assembly of new synapses and improved memory upon environmental enrichment.

Learning is correlated with the assembly of new synapses, but the roles of synaptogenesis processes in memory are poorly understood. Here, we show that mice lacking ß-Adducin fail to assemble new synapses upon enhanced plasticity and exhibit diminished long-term hippocampal memory upon environmental enrichment. Enrichment-enhanced the disassembly and assembly of dynamic subpopulations of synapses. Upon enrichment, stable assembly of new synapses depended on the presence of ß-Adducin, disassembly involved ß-Adducin phosphorylation through PKC, and both were required for augmented learning. In the absence of ß-Adducin, enrichment still led to an increase in spine structures, but the assembly of synapses at those spines was compromised. Virus-mediated re-expression of ß-Adducin in hippocampal granule cells of ß-Adducin(-/-) mice rescued new synapse assembly and learning upon enrichment. Our results provide evidence that synapse disassembly and the establishment of new synapses are both critically important for augmented long-term learning and memory upon environmental enrichment.

EphA4 signaling in juveniles establishes topographic specificity of structural plasticity in the hippocampus.

The formation and loss of synapses is involved in learning and memory. Distinct subpopulations of permanent and plastic synapses coexist in the adult brain, but the principles and mechanisms underlying the establishment of these distinctions remain unclear. Here we show that in the hippocampus, terminal arborizations (TAs) with high plasticity properties are specified at juvenile stages, and account for most synapse turnover of adult mossy fibers. Out of 9-12 giant terminals along CA3, distinct subpopulations of granule neurons revealed by mouse reporter lines exhibit 0, 1, or >2 TAs. TA specification involves a topographic rule based on cell body position and EphA4 signaling. Upon disruption of EphA4 signaling or PSA-NCAM in juvenile circuits, single-TA mossy fibers establish >2 TAs, suggesting that intra-axonal competition influences plasticity site selection. Therefore, plastic synapse specification in juveniles defines sites of synaptic remodeling in the adult, and hippocampal circuit plasticity follows unexpected topographic principles.

The plasma membrane calcium ATPase MCA-3 is required for clathrin-mediated endocytosis in scavenger cells of Caenorhabditis elegans.

Plasma membrane Ca2+ ATPases (PMCAs) maintain proper intracellular Ca2+ levels by extruding Ca2+ from the cytosol. PMCA genes and splice forms are expressed in tissue-specific patterns in vertebrates, suggesting that these isoforms may regulate specific biological processes. However, knockout mutants die as embryos or undergo cell death; thus, it is unclear whether other cell processes utilize PMCAs or whether these pumps are largely committed to the control of toxic levels of calcium. Here, we analyze the role of the PMCA gene, mca-3, in Caenorhabditis elegans. We report that partial loss-of-function mutations disrupt clathrin-mediated endocytosis in a class of scavenger cells called coelomocytes. Moreover, components of early endocytic machinery are mislocalized in mca-3 mutants, including phosphatidylinositol-4,5-bisphosphate, clathrin and the Eps15 homology (EH) domain protein RME-1. This defect in endocytosis in the coelomocytes can be reversed by lowering calcium. Together, these data support a function for PMCAs in the regulation of endocytosis in the C. elegans coelomocytes. In addition, they suggest that endocytosis can be blocked by high calcium levels.