Deep brain stimulation of the mediodorsal thalamic nucleus yields increases in the expression of zif-268 but not c-fos in the frontal cortex

This study explores the regions activated by deep brain stimulation of the mediodorsal thalamic nucleus through examination of immediate early genes as markers of neuronal activation. Stimulation was delivered unilaterally with constant current 100 μs duration pulses at a frequency of 130 Hz delivered at an amplitude of 200 μA for 3 h. Brains were removed, sectioned and radio-labelled for the IEGs zif-268 and c-fos. In anaesthetised rats, deep brain stimulation of mediodorsal thalamic nucleus produced robust increases in the expression of zif-268 but not c-fos localised to regions that are reciprocally connected with the mediodorsal thalamic nucleus, including the prelimbic and orbitofrontal cortices, and the premotor cortex indicating an increase in synaptic activity in these regions. These findings map those brain regions that are persistently, rather than transiently, activated by high frequency electrical stimulation of the mediodorsal thalamic nucleus by a putatively antidromic mechanism which may be relevant to neuropsychiatric disorders such as schizophrenia in which thalamocortical systems are disrupted and in which DBS protocols are being considered.     

Saliency-guided neural prosthesis for visual attention: Design and simulation

Recently the authors showed that a computational model of visual saliency could account for changes in gaze behavior of monkeys with damage in the primary visual cortex. Here we propose a neural prosthesis to restore eye gaze behavior by electrically stimulating the superior colliculus to drive visual attention. The saliency computational model is used to calculate the stimulation parameters from a real-time camera stream. Our simulations demonstrate that electrodes implanted in the superior colliculus at 1.0 mm spacing are, in principle, able to recover specifically those visual attention behaviors which are lost when the primary visual cortex is damaged.     

Effect of low frequency electrical stimulation on spike and wave discharges of perioral somatosensory cortex in WAG/Rij rats

Low frequency electrical stimulation has been revealed that as a potential cure in patient with drug resistant to epilepsy. This study tries to evaluate the effect of low frequency electrical stimulation (LFS) on absence seizure of perioral region primary somatosensory cortex (S1po). Eighteen male WAG/Rij rats were received LFS (3 Hz, square wave, monophasic, 200 μs, and 400 μA) for 25 min into S1po for a period of five days. There is 6 animals per group .The stimulating electrodes were implanted according to stereotaxic landmarks and EEG recording was obtained 30 min before and after LFS to analyse frequency, number and duration of spike–wave discharges (SWD). The results showed that in animals with unilateral stimulating electrodes (Exp1) in first and second days and also in animals with bilateral stimulating electrodes (Exp2) in days 3rd and 4th. LFS had significant decrease effects (p < 0.05) on mean number of SWD between pre-LFS. In comparison pre-LFS to post-LFS, mean of duration in Exp2 decreased significantly. In continuous application of LFS (5 days) only the data of first day was differently significant (p < 0.05) but data of other days had no difference. Comparison of data between Exp1, Exp2 and control groups showed that the mean number of Exp1 was significantly different (p < 0.05) and mean pick frequency in Exp2 was significantly decreased in comparison with Exp1 group (p < 0.05). The LFS of S1po produces significant antiepileptic effect on absence seizure but it was not persistent till the next day and shows a short time effect.     

Cortical activation during temporal preparation assessed by transcranial magnetic stimulation

Preparatory modulations relative to the timing of upcoming stimuli may involve activation or suppression mechanisms. Here, we assessed the interplay between these mechanisms with transcranial magnetic stimulation (TMS) of the motor cortex. Single- or paired-pulse TMS with 3- or 15-ms interstimulus intervals was delivered during the interval between the warning and the imperative stimuli (i.e., the foreperiod) of a choice reaction time task. Temporal uncertainty was manipulated through between-block variation of the foreperiod duration (500 or 2500 ms). The shortening of reaction time for the short foreperiod was accompanied with a decrease in amplitude of the single-pulse motor evoked potential (MEP), indicating corticospinal suppression. The co-occurring increase in amplitude of both paired-pulse MEPs (3 and 15 ms) expressed relative to single-pulse MEPs reveals released short intracortical inhibition (SICI) and enhanced intracortical facilitation (ICF). These results suggest that temporal preparation is associated with both corticospinal suppression and cortical activation.     

Facilitatory effect of paired-pulse stimulation by transcranial magnetic stimulation with biphasic wave-form

Transcranial magnetic stimulation (TMS) is used to probe corticospinal excitability by stimulating the motor cortex. Our aim was to enhance the effects of biphasic TMS by coupling a suprathreshold test pulse and a following subthreshold priming pulse to induce short-interval intracortical facilitation (SICF), which is conventionally produced with monophasic TMS. Biphasic TMS could potentially induce the SICF effect with better energy-efficiency and with lower stimulus intensities. This would make the biphasic paired-pulses better applicable in patients with reduced cortical excitability.A prototype stimulator was built to produce biphasic paired-pulses. Resting motor thresholds (rMTs) from the right and left hand abductor pollicis brevis muscles, and the right tibialis anterior muscle of eight healthy volunteers were determined using single-pulse paradigm with neuronavigated TMS. The rMTs and MEPs were measured using single-pulses and three paired-pulse setups (interstimulus interval, ISI of 3, 7 or 15 ms).The rMTs were lower and MEPs were higher with biphasic paired-pulses compared to single-pulses. The SICF effect was greatest at 3 ms ISI. This suggests that the application of biphasic paired-pulses to enhance stimulation effects is possible.     

Effect of low estrogen on neurons in the preoptic area of hypothalamus of ovariectomized rats

The purpose of this study was to investigate the difference in neuronal activity in the preoptic area of the hypothalamus (POAH) under low estrogen condition induced by ovariectomy. One hundred and twenty sham-operated (SHAM) and ovariectomized (OVX) rats were placed in different temperatures for 2 h. Twelve rats from each group were stimulated by 4 °C, 10 °C, 25 °C, 33 °C and 38 °C, respectively. c-Fos expression in the POAH was detected by immunohistochemistry. Following exposure to warm and cold stimuli, there were markedly lower c-Fos-positive cell densities in the OVX group compared with the SHAM group in the median preoptic nucleus (MnPO) at 4 °C, 10 °C, 33 °C and 38 °C, in the medial preoptic area (MPA) at 25 °C and 38 °C, in the ventromedial preoptic nucleus (VMPO) at 4 °C, 10 °C and 38 °C and in the ventrolateral preoptic nucleus (VLPO) at 4 °C and 38 °C. Both temperature and surgery had an impact on c-Fos expression by two-way ANOVA method except in the lateral preoptic area (LPO). c-Fos expression differed within different nuclei of the two groups in the same and different temperature stimuli. This indicated that the temperature-sensitive nuclei in the POAH exhibited lower and different activities during temperature stimuli following ovariectomy, which possibly resulted in abnormal thermoregulation and menopausal symptoms.     

The suitability of rat hepatoma cell line H4IIE for evaluating the potentials of compounds to induce CYP3A23 expression

To investigate the suitability of H4IIE cells for detecting cytochrome P450 (CYP) induction in vitro, we compared CYP induction by typical CYP inducers in H4IIE cells and rat primary hepatocytes by examining gene expression and enzyme activity, and by immunocytochemistry. The cells were preincubated with 0.1 μM of dexamethasone (DEX) for 24 h, followed by 48 h of exposure to 10 μM of beta-naphthoflavone (bNF), 100 μM of phenobarbital (PB) and 10 μM of DEX. Cyp1a1, Cyp2b1/2 and Cyp3a23/3a1 (Cyp3a23) expressions in H4IIE cells were up-regulated 280-, 1.5- and 65-fold relative to those in vehicle-treated cells, respectively. The fold inductions of those expressions in rat primary hepatocytes were 80-, 33- and 152-fold, respectively. Comprehensive gene expression analysis using DNA microarrays showed that Cyp3a23, Gsta2, Ugt2b12, Udpgt and Sult2a1 expressions were up-regulated in H4IIE cells exposed to 10 μM of DEX. CYP3A activity was not increased, but some H4IIE cells exposed to DEX were stained strongly with anti-CYP3A antibody. We cloned these cells and obtained cloned H4IIE (cH4IIE) cells with expression level of Cyp3a23 higher than those of vehicle-treated cells. It was confirmed that preincubation with 0.1 μM of DEX increased pregnane X receptor (Pxr) expression level and enhanced the Cyp3a23 induction effects of test compounds significantly. Retrospective examination of in vitro CYP induction assay using cH4IIE cells resulted in 80% correlation with the data from in vivo rat toxicity studies. These results suggested that cH4IIE cells are suitable for evaluating the potentials of a compound to induce CYP3A23 expression.     

Poly(3,4-ethylenedioxythiophene)/multiwall carbon nanotube composite coatings for improving the stability of microelectrodes in neural prostheses applications

With the purpose of improving the stability of microelectrodes under continuous high charge density stimulation, which is required for neural prostheses applications such as visual prostheses, multiwall carbon nanotube (MWCNT)-doped poly(3,4-ethylenedioxythiophene) (PEDOT) composite films were coated onto a platinum microelectrode by electrochemical polymerization. Galvanostatically polymerized PEDOT/MWCNT films demonstrated superior characteristics compared to polystyrene sulfonate doping and potentiostatic polymerization, including a three-dimensional cone morphology and enhanced electrochemical performance (the safe charge injection limit reached 6.2 mC cm^?2 for cathodic-first pulses). Most important of all, the improved stability of the coatings has been revealed through stimulation for 96 h using 3.0 mC cm^?2 current pulses in bicarbonate- and phosphate-buffered saline solution. Cell assays revealed that PEDOT/MWCNT films could promote the adhesion and neurite outgrowth of rat pheochromocytoma cells. Finally, platinum wires coated with PEDOT/MWCNT films were implanted into rat cortex for 6 weeks for histological evaluation. Glial fibrillary acidic protein and neuronal nuclei staining revealed that the films elicit a lower tissue response compared to platinum implants. These results suggest that the galvanostatically polymerized PEDOT/MWCNT films can improve the stability of stimulation microelectrodes and that PEDOT/MWCNT is an excellent candidate material for electrode coating for neural prostheses applications.     

Micropatterning–retinoic acid co-control of neuronal cell morphology and neurite outgrowth

Creating physical–biochemical superposed microenvironments optimal for stimulating neurite outgrowth would be beneficial for neuronal regenerative medicine. We investigated potential co-regulatory effects of cell micropatterning and retinoic acid (RA) soluble factor on neuronal cell morphology and neurite outgrowth. Human neuroblastoma (SH-SY5Y) cell patterning sensitivity could be enhanced by poly-l-lysine-g-polyethylene glycol cell-repellent back-filling, enabling cell confinement in lanes as narrow as 5 μm. Cells patterned on narrow (5 and 10 μm) lanes showed preferred nucleus orientation following the patterning direction. These cells also showed high nucleus aspect ratio but constrained nucleus spreading. On the other hand, cells on wide (20 μm and above) lanes showed random nucleus orientation and cell and nucleus sizes similar to those on unpatterned controls. All these changes were generally maintained with or without RA. Confining cells on narrow (5 and 10 μm) lanes, even without RA, significantly enhanced neurite extension relative to unpatterned control, which was further stimulated by RA. Interestingly, cell patterning on 5 and 10 μm lanes without RA produced longer neurites relative to the RA treatment alone case. Our data on the potential interplay between microscale physical cell confinement and RA-soluble stimulation may provide a new, integrative insight on how to trigger neurite/axon formation for neuronal regenerative medicine.     

Conducting polymers on hydrogel-coated neural electrode provide sensitive neural recordings in auditory cortex

Recently, a significant amount of effort has been dedicated to understanding factors that influence the functionality of bio-electronic sensors and to development of novel coating technologies for modifying biosensor surfaces. Due to its well-known biocompatibility, alginate hydrogel (HG) has been used as a coating material on neural electrodes for promoting intimate cellular integration, providing a scaffold for local drug delivery, and creating a mechanical buffer between hard electrodes and the soft tissues of the central nervous system. However, neural signal recordings using HG-coated electrodes in animal models are still poorly evaluated. Here, we investigated the effect of the proximity of source neurons around the electrode sites using HG coatings with various thicknesses deposited on microfabricated electrodes, implanted in auditory cortex of guinea pigs. We also evaluated the role of the conducting polymer poly(3,4-ethylenedioxythiophene) (PEDOT) in improving the recording functionality of the HG-coated neural electrodes. A significant loss in recording functionality was observed with thicker HG coatings, as determined by the number of clearly detectable units (30% with 80 μm thick coatings) and average signal-to-noise ratios (3.91 with 80 μm thick coatings). However, deposition of the conducting polymer PEDOT on the electrode sites restored the lost functionality of the electrodes caused by the HG coatings (30 μm). These conducting polymer/HG coatings have the potential to improve long-term performance of the neural electrodes not only by improving the electrode biocompatibility but also by facilitating more efficient signal transmission.     

The effect of FES of the tibial nerve on physiological activation of leg muscles during gait

The effects of surface functional electrical stimulation (FES) of the tibial nerve of healthy subjects were evaluated. The FES was applied at three different times during gait: early, mid and late stances. The purpose of this work is to understand the effect of unilateral stimulation on the bilateral activation patterns of leg muscles, because FES is used in practice to improve gait, while associated neuromuscular change is not often measured. The experimental protocol presented here will be transferred to stroke subjects, who could benefit from improved push-off during gait. Results show that FES of the tibial nerve changes the offset timing of the tibialis anterior muscle on the stimulated side and the on- and offset timings of the tibialis anterior muscle of the leg contralateral to stimulation. Additionally, activity levels of the semitendinosus ipsilateral and tibialis anterior contralateral to the stimulated leg significantly decreased, with respect to the non-stimulated condition. For the semitendinosus, this was a difference of 6–7 μV, with p < 0.05. For the tibialis anterior, this was a difference of 7–15 μV, with a significance of p = 0.00, respectively.This information is important for future applications of stimulation as it means that stimulation not only affects the stimulated muscle but also the physiological motor control by the CNS.     

Irisin protects against neuronal injury induced by oxygen-glucose deprivation in part depends on the inhibition of ROS-NLRP3 inflammatory signaling pathway

Recent studies found that irisin, a newly discovered skeletal muscle-derived myokine during exercise, is also synthesized in various tissues of different species and protects against neuronal injury in cerebral ischemia. The NOD-like receptor pyrin 3 (NLRP3) inflammasome play an important role in detecting cellular damage and mediating inflammatory responses to aseptic tissue injury during ischemic stroke. However, it is unclear whether irisin is involved in the regulation of NLRP3 inflammasome activation during ischemic stroke. In the present study, PC12 neuronal cells were exposed to oxygen-glucose deprivation (OGD), exogenous irisin (12.5, 25, 50 nmol/L) or NLRP3 inhibitor glyburide (50, 100, 200 μmol/L) were used as an intervention reagent, NLRP3 was over-expressed or suppressed by transfection with a NLRP3 expressing vector or NLRP3-specifc siRNA, respectively. Our data showed that both irisin and its precursor protein fibronectin type III domain containing 5 (FNDC5) expression were significantly down-regulated (p < 0.05); but oxidative stress and ROS-NLRP3 inflammasome signaling were activated by OGD (p < 0.05); treatment with irisin or inhibition of NLRP3 reversed OGD-induced oxidative stress and inflammation (p < 0.05). However, these irisin-mediated effects were blunted by over-expression NLRP3 (p < 0.05). Taken together, our results firstly revealed that irisin mitigated OGD-induced neuronal injury in part via inhibiting ROS-NLRP3 inflammatory signaling pathway, suggesting a likely mechanism for irisin-induced therapeutic effect in ischemic stroke.     

Metabotropic glutamate receptor agonists protect from oxygen–glucose deprivation- and colchicine-induced apoptosis in primary cultures of cerebellar granule cells

The effects of the metabotropic glutamate receptor agonists against apoptosis induced by oxygen–glucose deprivation or colchicine were studied in the primary cultures of mature cerebellar granule cells. Oxygen–glucose deprivation (90 min) or addition of colchicine (1 μM) resulted in neuronal damage as revealed by Trypan Blue assay 12 h later. Further analysis demonstrated that the cells exposed to oxygen–glucose deprivation or colchicine exhibit typical features of apoptosis: internucleosomal DNA fragmentation, condensation and fragmentation of chromatin and typical DNA ladder on agarose gel electrophoresis. Metabotropic glutamate receptor agonist, (1S,3R)-1-aminocycloheptane-trans-1,3-dicarboxylic acid, acting at group I and II receptors, and selective agonist, (2S,1′R,2R′,3R′)-2(2,3-dicarboxycyclopropyl)glycine, acting at group II receptors, added to cells recovering from oxygen-glucose deprivation exerted neuroprotective action against oxygen–glucose deprivation-induced apoptosis. Similar neuroprotective effects of metabotropic glutamate receptor agonists were observed against colchicine-induced apoptosis. The results thereby provide evidence that metabotropic glutamate receptor agonists have therapeutic potential in the treatment of pathologies associated with increased neuronal apoptosis. The selective protein kinase C inhibitor bisindolylmaleimide (100 nM) abolished the neuroprotective action of (1S,3R)-1-aminocycloheptane-trans-1,3-dicarboxylic acid, whereas the activator of adenylyl cyclase forskolin (10 μM) abolished the neuroprotective action of (2S,1′R,2R′,3R′)-2(2,3-dicarboxycyclopropyl)glycine (30 μM) against colchicine-induced apoptosis.It is concluded that both phosphoinositide hydrolysis with consequent activation of protein kinase C and inhibition of adenylyl cyclase seem to contribute to the neuroprotective action of metabotropic glutamate receptor agonists against neuronal apoptosis in the primary culture of cerebellar granule cells.     

Sodium and water intake are not affected by GABAC receptor activation in the lateral parabrachial nucleus of sodium-depleted rats

The activation of GABAergic receptors, GABA_A and GABA_B, in the lateral parabrachial nucleus (LPBN) increases water and sodium intake in satiated and fluid-depleted rats. The present study investigated the presence of the GABA_C receptor in the LPBN, its involvement in water and sodium intake, and its effects on cardiovascular parameters during the acute fluid depletion induced by furosemide combined with captopril (Furo/Cap). One group of male Wistar rats (290–300 g) with bilateral stainless steel LPBN cannulas was used to test the effects of a GABA_C receptor agonist and antagonist on the fluid intake and cardiovascular parameters. We investigated the effects of bilateral LPBN injections of trans-4-aminocrotonic acid (TACA) on the intake of water and 0.3 M NaCl induced by acute fluid depletion (subcutaneous injection of Furo/Cap). c-Fos expression increased (P<0.05), suggesting LPBN neuronal activation. The injection of different doses of TACA (0.5, 2.0 and 160 nmol) in the LPBN did not change the sodium or water intake in Furo/Cap-treated rats (P > 0.05). Treatment with the GABA_C receptor antagonist (Z)-3-[(aminoiminomethyl)thio]prop-2-enoic acid sulfate (ZAPA, 10 nmol) or with ZAPA (10 nmol) plus TACA (160 nmol) did not change the sodium or water intake compared with that for vehicle (saline) (P > 0.05). Bilateral injections of the GABA_C agonist in the LPBN of Furo/Cap-treated rats did not affect the mean arterial pressure (MAP) or heart rate (HR). The GABA_C receptor expression in the LPBN was confirmed by the presence of a 50 kDa band. Although LPBN neurons might express GABA_C receptors, their activation produced no change in water and sodium intake or in the cardiovascular parameters in the acute fluid depletion rats. Therefore, the GABA_C receptors in the LPBN might not interfere with fluid and blood pressure regulation.     

Size-controlled insulin-secreting cell clusters

The search for an effective cure for type I diabetes from the transplantation of encapsulated pancreatic β-cell clusters has so far produced sub-optimal clinical outcomes. Previous efforts have not controlled the size of transplanted clusters, a parameter implicated in affecting long-term viability and the secretion of therapeutically sufficient insulin. Here we demonstrate a method based on covalent attachment of patterned laminin for fabricating uniformly size-controlled insulin-secreting cell clusters. We show that cluster size within the range 40–120 μm in diameter affects a variety of therapeutically relevant cellular responses including insulin expression, content and secretion. Our studies elucidate two size-dependent phenomena: (1) as the cluster size increases from 40 μm to 60 μm, glucose stimulation results in a greater amount of insulin produced per cell; and (2) as the cluster size increases beyond 60 μm, sustained glucose stimulation results in a greater amount of insulin secreted per cell. Our study describes a method for producing uniformly sized insulin-secreting cell clusters, and since larger cluster sizes risk nutrient availability limitations, our data suggest that 100–120 μm clusters may provide optimal viability and efficacy for encapsulated β-cell transplants as a treatment for type I diabetes and that further in vivo evaluation is warranted.     

Stronger Toll-like receptor 1/2, 4, and 7/8 but less 9 responses in peripheral blood mononuclear cells in non-infectious exacerbated asthmatic children

Toll-like receptors (TLR) initiate innate and often affect adaptive immune response. This study aimed to determine if TLR response and T regulatory cell (Treg) function in peripheral blood mononuclear cells (PBMC) correlate with clinical severity in non-infectious asthma. TLR1–9 expression and representative response cytokine TNF-α, IL-6, and IFN-β secretions were analyzed after stimulation by TLR1–9 ligands from 17 non-infectious asthmatic children. TNF-α production was higher in TLR1/2 (median 385.4 vs. 250.3 pg/ml in 1 μg/ml Pam3CSK4, p = 0.0078), TLR4 (2392.4 vs. 1355.9 in 1 μg/ml LPS; p = 0.0005), and TLR7/8 (10,776.2 vs. 4237.0 pg/ml in 1 μg/ml R848, p = 0.0079) of patients in exacerbation than those in convalescence and healthy controls despite equal TLR expression. TNF-α production stimulated by TLR9 agonist was significantly lower in exacerbation (17.7 vs. 34.9 pg/ml in 1 μg/ml ODN2216, p = 0.0175), while IL-6 production had similar patterns but was significantly lower in TLR3 signaling (119.7 vs. 245.0 pg/ml in 0.1 μg/ml poly(I:C), p = 0.0033). IFN-β production by TLR3 agonist also decreased in exacerbation but not statistically significant. Six older children showed decreased FOXP3 percentage in CD4 + CD25^high and decreased suppression capability in exacerbation but restored in stabilization (82.8% vs. 90.0%, p = 0.0061 and 60.9% vs. 81.7%, p = 0.0071; respectively). In conclusion, normalizing imbalanced TLR signaling and enhancing Treg cell capability may guide possible therapeutic strategies for non-infectious asthma in exacerbation.     

Parvalbumin expression in the claustrum of the adult dog. An immunohistochemical and topographical study with comparative notes on the structure of the nucleus

Although the detailed structure and function of the claustrum remain enigmatic, its extensive reciprocal connection with the cortex suggests a role in the integration of multisensory information.Claustrum samples, obtained from necropsy of four dogs, were formalin fixed for paraffin embedding. Sections were either stained for morpho-histological analysis or immunostained for parvalbumin (PV). We focused on PV because in cortical and hippocampal areas it is a marker of the fast-spiking interneurons which have an important role in the information transmission and processing. Soma area, perimeter and circularity were considered as morphological parameters to quantitatively group the PV positive somata by k-means clustering.The histological investigation revealed a superior pyramidoid puddle and a posterior puddle characterized by a “cloud” of neurons in its dorso-lateral part. Immunostaining showed positive somata and fibers throughout the rostro-caudal extent of the dog claustrum, localized principally in the dorsal region. k-Means clustering analysis enabled neuron classification according to size, identifying respectively big (radius = 11.42 ± 1.99 μm) and small (radius = 6.33 ± 1.08 μm) cells. No statistical differences in soma shape were observed. The topographical distribution of PV immunoreactivity suggests that the dog dorsal claustrum might be functionally related to the processing of visual inputs.Taken together our findings may help in the understanding the physiology of claustrum when compared with anatomical and functional data obtained in other species.     

In vitro studies of the impact of maribavir on CMV-specific cellular immune responses

BackgroundGanciclovir has demonstrated immunosuppressive effects in vitro which may lead to delayed cytomegalovirus (CMV)-specific immune reconstitution when the drug is given prophylactically. Maribavir is a new and more potent anti-CMV drug that is under evaluation for therapeutic use in transplant recipients.ObjectivesThe objective of this study was to evaluate the potential effect of maribavir on CMV-specific T cell function in comparison to ganciclovir.Study designIn ten immunocompetent CMV seropositive donors, maribavir and ganciclovir were compared over a broad range of concentrations (0.2–500 μM) regarding their effects on lymphoproliferation, CMV-specific CD4+ and CD8+ cytokine expression, T cell multifunctionality, degranulation and apoptosis.ResultsMaribavir inhibited lymphocyte proliferation at concentrations of 50 μM and above, however, cytokine expression, cellular degranulation and multifunctionality of CD4+ and CD8+ T cells in response to CMV lysate and pp65 peptide mix were not impaired except at the highest concentration of 500 μM. Ganciclovir inhibited lymphoproliferative responses starting at 10 μM. As with maribavir, other cellular responses following stimulation with CMV lysate and pp65 peptide mix were only impaired at the highest concentration of 500 μM of ganciclovir. Neither maribavir nor ganciclovir showed induction of lymphocyte apoptosis.ConclusionsMaribavir exhibits a low potential to suppress CMV-specific T cell function. This finding supports the use of higher doses in the prophylactic setting than originally proposed.     

Automated setup of functional electrical stimulation for drop foot using a novel 64 channel prototype stimulator and electrode array: Results from a gait-lab based study

Functional electrical stimulation is commonly used to correct drop foot following stroke or multiple sclerosis. This technique is successful for many patients, but previous studies have shown that a significant minority have difficulty identifying correct sites to place the electrodes in order to produce acceptable foot movement. Recently there has been some interest in the use of ‘virtual electrodes’, the process of stimulating a subset of electrodes chosen from an array, thus allowing the site of stimulation to be moved electronically rather than physically. We have developed an algorithm for automatically determining the best site of stimulation and tested it on a computer linked to a small, battery-powered prototype stimulator with 64 individual output channels. Stimulation was delivered via an 8 × 8 array adhered to the leg by high-resistivity self-adhesive hydrogel. Ten participants with stroke (ages 53–71 years) and 11 with MS (ages 40–80 years) were recruited onto the study and performed two walks of 10 m for each of the following conditions: own setup (PS), clinician setup (CS), automated setup (AS) and no stimulation (NS). The PS and CS conditions used the participant's own stimulator with two conventional electrodes; the AS condition used the new stimulator and algorithm. Outcome measures were walking speed, foot angle at initial contact and the Borg Rating of Perceived Exertion.Mean walking speed with no stimulation was 0.61 m/s; all FES setups significantly increased speed relative to this (AS p < 0.05, PS p < 0.01, CS p < 0.01). Speed for PS (0.72 m/s) was faster than both AS (0.65 m/s, p < 0.01) and CS (0.68 m/s, p < 0.05). Frontal plane foot orientation at heel-strike was more neutral for AS (0.3° everted) than in the NS (11.2° inverted, p < 0.01), PS (4.5° inverted, p < 0.05) and CS (3.1° inverted, p < 0.05) conditions. Dorsiflexion angles for AS (4.2°) were larger than NS (?3.0°, p < 0.01), not different to PS (4.3°, p > 0.05) and less dorsiflexed than CS (6.0°, p < 0.05).This proof of principle study has demonstrated that automated setup of an array stimulator produces results broadly comparable to clinician setup. Slower walking speed for automated and clinician setups compared to the participants’ own setup may be due to the participants’ lack of familiarity with responses different to their usual setups. Automated setup using the method described here seems sufficiently reliable for future longer-term investigation outside the laboratory and may lead to FES becoming more viable for patients who, at present, have difficulty setting up conventional stimulators.     

Neurochemical study of amino acids in rodent brain structures using an improved gas chromatography–mass spectrometry method

The analysis of amino acid levels is crucial for neuroscience studies because of the roles of these molecules as neurotransmitters and their influence on behavior. The present study describes the distribution and levels of 16 amino acids (alanine, asparagine, aspartic acid, cysteine, glycine, glutamic acid, isoleucine, leucine, lysine, methionine, phenylalanine, proline, sarcosine, serine, valine, and threonine) in brain tissues (prefrontal cortex, striatum, hippocampus and cerebellum) and the serum. Neurochemical analysis was performed on Wistar rats and C57BL/6 mice using an efficient method for extraction, a fast microwave-assisted derivatization and gas chromatography–mass spectrometry analysis. The amino acid concentration varied across brain regions for 14 of the 16 analyzed molecules, with detection limits ranging from 0.02 ± 0.005 μmol L⁻¹ to 7.07 ± 0.05 μmol L⁻¹. In rats, the concentrations of alanine, glycine, methionine, serine and threonine were higher in prefrontal cortex than in other areas, whereas in mice, the concentrations of glutamic acid, leucine and proline were highest in the hippocampus. In conclusion, this study provides a cerebral profile of amino acids in brain regions and the serum of rats and mice.