Murine epidermal Langerhans cells do not express inducible nitric oxide synthase

In Leishmania-infected macrophages (MΦ), the formation of reactive nitrogen intermediates by the inducible isoform of nitric oxide synthase (iNOS) is critical for the killing of the intracellular parasites. We have recently shown that, in addition to MΦ, epidermal Langerhans cells (LC) can phagocytose Leishmania major, but they do not allow parasite replication. Therefore, we analyzed whether LC and MΦ display the same leishmanicidal effector mechanism. Unlike MΦ, stimulation of unselected epidermal cells with interferon-γ/lipopoly-saccharide did not lead to the release of nitric oxide (NO), and inhibition of NO production had no effect on the rate of infection of LC. iNOS mRNA was clearly detectable in MΦ as well as unselected epidermal cells (the majority of which consists of keratinocytes) after stimulation with different cytokines. In contrast, pure LC obtained by single-cell picking from cytokine-activated or L. major-infected epidermal cells did not express iNOS mRNA. Addition of the NO donor S-nitroso-N-acetylpenicillamine to already-infected LC did not alter their rate of infection, indicating that LC do not utilize exogenous NO for the control of intracellular Leishmania. These results suggest that in the L. major-infected skin, activated MΦ and keratinocytes, but not LC have the ability to express iNOS activity. Therefore, an as yet unidentified, NO-independent mechanism appears to be responsible for the control of parasite replication in LC.     

Streptococcus pneumoniae exposure is associated with human metapneumovirus seroconversion and increased susceptibility to in vitro HMPV infection

It remains largely unknown which factors determine the clinical outcome of human metapneumovirus (HMPV) infections. The aim of the present study was to analyse whether exposure to bacterial pathogens can influence HMPV infections. From 57 children, serum samples and colonization data for Haemophilus influenzae, Moraxella catarrhalis, Staphylococcus aureus and Streptococcus pneumoniae were collected at 1.5, 6, 14 and 24 months of age. Seroconversion rates to HMPV were determined and related to bacterial carriage. Frequent nasopharyngeal carriage (≥2 times in the first 2 years of life) of S. pneumoniae, but not of the other three pathogens, was associated with increased seroconversion rates of infants to HMPV at the age of 2 years (frequently vs. less exposed, 93% vs. 59%; p <0.05). Subsequently, the susceptibility of well-differentiated normal human bronchial epithelial cells (wd-NHBE) pre-incubated with bacterial pathogens to in vitro HMPV infection was evaluated. Pre-incubation of wd-NHBE with S. pneumoniae resulted in increased susceptibility to infection with HMPV-enhanced green fluorescent protein (EGFP), as determined by enumeration of EGFP-positive cells. This was not the case for cells pre-incubated with H. influenzae, M. catarrhalis on S. aureus. We conclude that exposure to S. pneumoniae can modulate HMPV infection.     

Acute respiratory distress syndrome leads to reduced ratio of ACE/ACE2 activities and is prevented by angiotensin-(1-7) or an angiotensin II receptor antagonist

Acute respiratory distress syndrome (ARDS) is a devastating clinical syndrome. Angiotensin-converting enzyme (ACE) and its effector peptide angiotensin (Ang) II have been implicated in the pathogenesis of ARDS. A counter-regulatory enzyme of ACE, ie ACE2 that degrades Ang II to Ang-(1-7), offers a promising novel treatment modality for this syndrome. As the involvement of ACE and ACE2 in ARDS is still unclear, this study investigated the role of these two enzymes in an animal model of ARDS. ARDS was induced in rats by intratracheal administration of LPS followed by mechanical ventilation. During ventilation, animals were treated with saline (placebo), losartan (Ang II receptor antagonist), or with a protease-resistant, cyclic form of Ang-(1-7) [cAng-(1-7)]. In bronchoalveolar lavage fluid (BALF) of ventilated LPS-exposed animals, ACE activity was enhanced, whereas ACE2 activity was reduced. This was matched by enhanced BALF levels of Ang II and reduced levels of Ang-(1-7). Therapeutic intervention with cAng-(1-7) attenuated the inflammatory mediator response, markedly decreased lung injury scores, and improved lung function, as evidenced by increased oxygenation. These data indicate that ARDS develops, in part, due to reduced pulmonary levels of Ang-(1-7) and that repletion of this peptide halts the development of ARDS.     

NKG2D performs two functions in invariant NKT cells: direct TCR-independent activation of NK-like cytolysis and co-stimulation of activation by CD1d

Invariant NKT cells are important in the activation and regulation of immune responses. They can also function as CD1d-restricted killer cells. However, the role of activating innate NK-cell receptors expressed on NKT cells in triggering cytolytic function is poorly characterized. Here, we initially confirmed that the cellular stress-ligand receptor NKG2D is expressed on CD4- NKT cells, whereas most CD4+ NKT cells lack this receptor. Interestingly, NKG2D+ NKT cells frequently expressed perforin, and both NKG2D and perforin localized at the site of contact with NKG2D ligand-expressing target cells. CD4- NKT cells degranulated in response to NKG2D engagement in a redirected activation assay independent of stimulation via their invariant TCR. NKT cells killed P815 cells coated with anti-NKG2D mAb and CD1d-negative K562 tumor target cells in an NKG2D-dependent manner. Furthermore, NKG2D engagement co-stimulated TCR-mediated NKT-cell activation in response to endogenous CD1d-presented ligands or suboptimal levels of anti-CD3 triggering. These data indicate that the CD4- subset of human NKT cells can mediate direct lysis of target cells via NKG2D engagement independent of CD1d, and that NKG2D also functions as a co-stimulatory receptor in these cells. NKG2D thus plays both a direct and a co-stimulatory role in the activation of NKT cells.     

Combined spiroergometry and 31P–MRS of human calf muscle during high‐intensity exercise

Simultaneous measurements of pulmonary oxygen consumption (VO₂), carbon dioxide exhalation (VCO₂) and phosphorus magnetic resonance spectroscopy (³¹P–MRS) are valuable in physiological studies to evaluate muscle metabolism during specific loads. Therefore, the aim of this study was to adapt a commercially available spirometric device to enable measurements of VO₂ and VCO₂ whilst simultaneously performing ³¹P–MRS at 3 T. Volunteers performed intense plantar flexion of their right calf muscle inside the MR scanner against a pneumatic MR‐compatible pedal ergometer. The use of a non‐magnetic pneumotachograph and extension of the sampling line from 3 m to 5 m to place the spirometric device outside the MR scanner room did not affect adversely the measurements of VO₂ and VCO₂. Response and delay times increased, on average, by at most 0.05 s and 0.79 s, respectively. Overall, we were able to demonstrate a feasible ventilation response (VO₂ = 1.05 ± 0.31 L/min; VCO₂ = 1.11 ± 0.33 L/min) during the exercise of a single calf muscle, as well as a good correlation between local energy metabolism and muscular acidification (τ_{PCr fast} and pH; R² = 0.73, p < 0.005) and global respiration (τ_{PCr fast} and VO₂; R² = 0.55, p = 0.01). This provides improved insights into aerobic and anaerobic energy supply during strong muscular performances.     

Interplay of neuronal processes during response inhibition: Results from a combined event-related potentials (ERPs)/transcranial magnetic stimulation (TMS) study on methylphenidate

The neuronal processes underlying response inhibition are often studied using either event-related potentials (ERPs) or by applying transcranial magnetic stimulation (TMS) to investigate excitatory and inhibitory processes in the motor system. We performed a more refined analysis of response inhibition by combining both approaches with the aim of identifying an interplay between ERPs and TMS parameters.During a go/nogo task, motor system excitability was measured using TMS single and double pulses and brain electrical activity was recorded in healthy adults (n = 14). Each participant completed two testing sessions, once on placebo and once on methylphenidate (double-blind, crossover design). Studying the effects of methylphenidate served as an example application for this combined approach.Developing regression models, inhibition-related TMS measures (e.g., short intracortical inhibition) and the contingent negative variation explained about 85% of the variance of the nogo-P3 under both MPH and placebo medication. The smaller the inhibitory effect in the motor system, the more terminal response control was required and the more resources were allocated for the evaluation of the inhibitory process, respectively, as indicated by a larger P3.Thus, an interplay between processes in the motor system (cortex) and control processes with sources in the prefrontal cortex and the anterior cingulate cortex (ACC) may take place, acting complementarily to facilitate a correct nogo-response.While ERPs rather represent initiation and monitoring of inhibitory processes and response control, motor inhibition may be best analyzed using TMS. A combined ERP/TMS analysis may allow for the development of distinct models concerning the interplay of processes involved in response inhibition.     

Attentional processes in children with ADHD: An event-related potential study using the attention network test

A variety of event-related potential (ERP) based studies have shown differences in neuronal processes underlying attention, inhibition and error processing in children with attention-deficit/hyperactivity disorder (ADHD) compared to controls. However, so far there are no studies that have compared children with ADHD and typically developing (TD) children regarding effects in ERP components associated with the attention network test (ANT). The ANT allows to differentiate between three particular aspects of attention: alerting, orienting, conflict.Twenty-five children with ADHD and 19 TD children (comparable with respect to age, sex, and IQ) performed the ANT while ERPs were recorded. Based on DSM-IV, the group of children with ADHD was divided in an inattentive (ADHDin, n = 10) and a combined (ADHDcom, n = 15) subgroup.On the performance level, the ADHD group showed a significantly higher variability of reaction times. Concerning ERP measures, smaller cue-P3 amplitudes were found in the ADHD group indicating that children with ADHD allocate less attentional resources for cue processing. In addition, the target-P3 in ADHD showed smaller amplitudes. Subgroup analysis revealed reduced cue-P3 amplitudes in both subgroups and reduced target-P3 amplitudes in ADHDin compared to TD children. Except for a higher alerting score in ADHD after correction for cue-P3 group differences, performance data revealed no group differences specific for the three attention networks. No group differences related to the attention networks were observed at the ERP level.Our results suggest that deviant attentional processing in children with ADHD is only partly related to ANT-specific effects. Findings are compatible with the model of a suboptimal energetic state regulation in ADHD. Furthermore, our results suggest that deviant cue processing in ADHD and related differences in task modulations should be accounted for in data analysis.     

Costes y adherencia del tratamiento antirretroviral

ObjectiveTo develop a system of data management that allows us to estimate the comparative effectiveness of the various antiretroviral treatment (ART) regimens.MethodRestrospective observational study in patients infected with HIV with stable ART. Adherence to treatment and unit cost for each patient's treatment was determined. The cost/patient/day was calculated and, multiplying by an adherence factor (f_ADH), the (cost/patient/day)_ADH. The comparison of both allowed us to obtain the Δ_cost/patient, which estimates the additional costs caused by lack of adherence. The incremental cost-effectiveness (iCER), grouping the results by the various coformulated drugs (“combos”). A study of the budgetary impact of these combos was carried out.Results468 patients were evaluated (62% adherent). Average adherence was 88±18%. The average value of (cost/patient/day) _ADH was significantly higher than the cost/patient/day (27.3 ± 9.8€ compared to 24.3±7.6€. p < 0.001). Just as with the f_ADH, no differences were found in the Δ_cost/patient between the different ART combinations. The combo with the least deviation from the cost/patient/day due to lack of adherence was that composed of abacavir/zedovudine/lamivudine (ABC/AZT/3TC,Δ_cost/patient=8.72±14.18%), and that with the greatest deviation AZT/3TC (Δ_cost/patient=13.52±17.68%). No significant differences were found in the iCER calcluated for any combo. The ART that included abacavir/lamivudine (ABC/3TC) obtained the least budgetary impact.ConclusionsThe greatest cost and percentage of adherent patients associated with the combos composed of Tenovovir/Emtricitabine(TDF/FTC) and ABC/3TC, and the least cost and effectiveness of those composed of AZT/#TC and ABC/AZT/3TC, does not allow us to identify any option as significantly dominant. The regimens with ABC/3TC were shown to be the most favourable from the combined point of view of cost and adherence.     

Symptomatic vertebral artery stent fracture: a case report

Percutaneous transluminal angioplasty (PTA) with stent placement is the preferred treatment modality at present for atherosclerotic stenotic lesions of vertebral artery origin. A complication of stent placement in the vertebral artery origin that has received little attention is the risk of stent fracture. A case with four-vessel pathology treated with PTA and stent placement in the left vertebral artery origin is presented. Symptoms recurred 4 months after stent placement, and arteriogram revealed a fractured stent, which was treated surgically with stent removal and vertebral artery-common carotid artery reimplantation.