From planetary to regional boundaries for agricultural nitrogen pollution.

Excessive agricultural nitrogen use causes environmental problems globally1, to an extent that it has been suggested that a safe planetary boundary has been exceeded2. Earlier estimates for the planetary nitrogen boundary3,4, however, did not account for the spatial variability in both ecosystems' sensitivity to nitrogen pollution and agricultural nitrogen losses. Here we use a spatially explicit model to establish regional boundaries for agricultural nitrogen surplus from thresholds for eutrophication of terrestrial and aquatic ecosystems and nitrate in groundwater. We estimate regional boundaries for agricultural nitrogen pollution and find both overuse and room for intensification of agricultural nitrogen. The aggregated global surplus boundary with respect to all thresholds is 43 megatonnes of nitrogen per year, which is 64 per cent lower than the current (2010) nitrogen surplus (119 megatonnes of nitrogen per year). Allowing the nitrogen surplus to increase to close yield gaps in regions where environmental thresholds are not exceeded lifts the planetary nitrogen boundary to 57 megatonnes of nitrogen per year. Feeding the world without trespassing regional and planetary nitrogen boundaries requires large increases in nitrogen use efficiencies accompanied by mitigation of non-agricultural nitrogen sources such as sewage water. This asks for coordinated action that recognizes the heterogeneity of agricultural systems, non-agricultural nitrogen losses and environmental vulnerabilities.

Medical students and physical education students as CPR instructors: an appropriate solution to the CPR-instructor shortage in secondary schools?

BACKGROUND: Integrating cardiopulmonary resuscitation (CPR) training in secondary schools will increase the number of potential CPR providers. However, currently too few certified instructors are available for this purpose. Training medical students and physical education student teachers to become CPR instructors could decrease this shortage. AIM: Examine whether medical students and physical education student teachers can provide CPR training for secondary school pupils as well as (i. e., non-inferior to) registered nurses. METHODS: A total of 144 secondary school pupils were randomly assigned to CPR training by a registered nurse (n = 12), a  medical student (n = 17) or a physical education student teacher (n = 15). CPR performance was assessed after training and after eight weeks in a simulated cardiac arrest scenario on a resuscitation manikin, using manikin software and video recordings. RESULTS: No significant differences were found between the groups on the overall Cardiff Test scores and the correctness of the CPR techniques during the post-training and retention test. All pupils showed sufficient CPR competence, even after eight weeks. CONCLUSION: Training by medical students or physical education student teachers is non-inferior to training by a registered nurse, suggesting that school teachers, student teachers and medical students can be recruited for CPR training in secondary schools.

Assessing nitrate groundwater hotspots in Europe reveals an inadequate designation of Nitrate Vulnerable Zones.

Monitoring networks show that the European Union Nitrates Directive (ND) has had mixed success in reducing nitrate concentrations in groundwater. By combining machine learning and monitored nitrate concentrations (1992-2019), we estimate the total area of nitrate hotspots in Europe to be 401,000 km2, with 47% occurring outside of Nitrate Vulnerable Zones (NVZs). We also found contrasting increasing or decreasing trends, varying per country and time periods. We estimate that only 5% of the 122,000 km2 of hotspots in 2019 will meet nitrate quality standards by 2040 and that these may be offset by the appearance of new hotspots. Our results reveal that the effectiveness of the ND is limited by both time-lags between the implementation of good practices and pollution reduction and an inadequate designation of NVZs. Substantial improvements in the designation and regulation of NVZs are necessary, as well as in the quality of monitoring stations in terms of spatial density and information available concerning sampling depth, if the objectives of EU legislation to protect groundwater are to be achieved.

Biomechanics of wheelchair turning manoeuvres: novel insights into wheelchair propulsion.

Introduction: Wheelchair turning biomechanics is an under researched area despite its obvious relevance to functional mobility of wheelchair users. Wheelchair turns might be linked to a higher risk of upper limb injuries due to the increased forces and torques potentially associated with asymmetric movement. Our aim was to obtain a better theoretical understanding of wheelchair turning by biomechanically analyzing turns compared to steady-state straightforward propulsion (SSSFP). Methods: Ten able-bodied men received 12-min familiarization and 10 trials (in a random order) of SSSFP and multiple left and right turns around a rectangular course. A Smartwheel was mounted at the right wheel of a standard wheelchair to measure kinetic parameters during SSSFP and of the inner hand during right turns and the outer hand during left turns. A repeated measures ANOVA was used to detect differences across tasks. Results: Two strategies were identified: 3% demonstrated roll turns and 97% spin turns. Spin turns consisted of three phases: approach, turning and depart phase. The turning phase was accomplished by increasing peak force (72.9 ± 25.1 N vs. 43.38 ± 15.9 N in SSSFP) of the inner hand, while maintaining high push frequency of the outer hand (1.09 ± 0.20 push/s vs. 0.95 ± 0.13 push/s in SSSFP). Peak negative force and force impulse during the turning phase were much higher than SSSFP, 15.3 ± 15.7 and 4.5 ± 1.7 times higher, respectively. Conclusion: The spin turn strategy might carry an increased risk of upper limb injuries due to higher braking force and requires particular attention by rehabilitation professionals to preserve upper limb function of long-term wheelchair users.

Large variations between NICU nurses in predicting nasogastric tube insertion length in a mannequin study.

Aim: To evaluate the inter- and intraindividual variation of predicted nasogastric tube insertion lengths by nurses working in two neonatal intensive care units in the Netherlands, using a mannequin model. Methods: A total of 110 nurses (55 nurses from Center A and 55 from Center B) were asked to predict the nasogastric tube insertion length on a neonatal mannequin. We evaluated the length and prediction method used by the nurses. We also estimated the number of tubes that would have correctly been placed in the stomach of a neonate according to the seize of the mannequin. Results: The mean predicted insertion length of the nasogastric tube was 30.0 cm with an interindividual variation of 12 cm (range 24-36 cm). The mean intraindividual variation was 0.75 cm. The two centers used two different prediction methods in their local guidelines, but overall at least 6 different methods were used by the nurses. We estimated that 77% (85/110) of the tubes would have ended in the body of the mannequins stomach, while 10% (11/110) would have ended in the esophagus and 13% (14/110) would have ended against the stomach lining or in the duodenum. Conclusion: Nurses in two neonatal intensive care units used many different methods which lead to a large interindividual variation in predicted insertion lengths of the nasogastric tubes. Regular evaluations using this mannequin model could lead to more uniformity and reduce the risk of tube misplacement in neonates.

Can we detect non-functional overreaching in young elite soccer players and middle-long distance runners using field performance tests?

OBJECTIVE: To study whether field performance tests can make a valid distinction between non-functionally overreaching (NFO) athletes and control athletes. DESIGN: Monthly field performance tests were used to determine a performance decrement (PD) throughout a season. Athletes with a minimum of 1 month PD were compared with control athletes without a PD on mood characteristics and resting levels of stress hormones. SETTING: Sporting field and sports medical laboratory. PARTICIPANTS: 129 young elite athletes, 77 soccer players and 52 middle-long distance runners were followed prospectively during the 2006-2007 season. Fifteen of them were invited to the laboratory. Eight athletes showed a performance decrease lasting longer than 1 month, and seven athletes without a performance decrease acted as their controls. MAIN OUTCOME MEASURES: Performance changes over time were measured using field tests. Profile of Mood States and resting levels of adrenocorticotrophic hormone (ACTH) and cortisol in blood were measured in the laboratory. RESULTS: PD athletes showed several symptoms typical of the non-functional state of overreaching (OR). The PD group scored higher on depression and anger than controls. They also showed a specific pattern of correlations between negative mood subscales (tension, fatigue and depression), which was absent in controls. ACTH levels at rest were similar, but lower cortisol levels in PD athletes pointed at a blunted cortisol response. Cortisol levels were decoupled from ACTH levels only in PD athletes. CONCLUSIONS: Implementing performance-related criteria in field tests can help coaches and sports physicians to distinguish NFO athletes from athletes with balanced workload and recovery.

Global assessment of nitrogen deposition effects on terrestrial plant diversity: a synthesis.

Atmospheric nitrogen (N) deposition is a recognized threat to plant diversity in temperate and northern parts of Europe and North America. This paper assesses evidence from field experiments for N deposition effects and thresholds for terrestrial plant diversity protection across a latitudinal range of main categories of ecosystems, from arctic and boreal systems to tropical forests. Current thinking on the mechanisms of N deposition effects on plant diversity, the global distribution of G200 ecoregions, and current and future (2030) estimates of atmospheric N-deposition rates are then used to identify the risks to plant diversity in all major ecosystem types now and in the future. This synthesis paper clearly shows that N accumulation is the main driver of changes to species composition across the whole range of different ecosystem types by driving the competitive interactions that lead to composition change and/or making conditions unfavorable for some species. Other effects such as direct toxicity of nitrogen gases and aerosols, long-term negative effects of increased ammonium and ammonia availability, soil-mediated effects of acidification, and secondary stress and disturbance are more ecosystem- and site-specific and often play a supporting role. N deposition effects in mediterranean ecosystems have now been identified, leading to a first estimate of an effect threshold. Importantly, ecosystems thought of as not N limited, such as tropical and subtropical systems, may be more vulnerable in the regeneration phase, in situations where heterogeneity in N availability is reduced by atmospheric N deposition, on sandy soils, or in montane areas. Critical loads are effect thresholds for N deposition, and the critical load concept has helped European governments make progress toward reducing N loads on sensitive ecosystems. More needs to be done in Europe and North America, especially for the more sensitive ecosystem types, including several ecosystems of high conservation importance. The results of this assessment show that the vulnerable regions outside Europe and North America which have not received enough attention are ecoregions in eastern and southern Asia (China, India), an important part of the mediterranean ecoregion (California, southern Europe), and in the coming decades several subtropical and tropical parts of Latin America and Africa. Reductions in plant diversity by increased atmospheric N deposition may be more widespread than first thought, and more targeted studies are required in low background areas, especially in the G200 ecoregions.

Use of dynamic soil-vegetation models to assess impacts of nitrogen deposition on plant species composition: an overview.

Field observations and experimental data of effects of nitrogen (N) deposition on plant species diversity have been used to derive empirical critical N loads for various ecosystems. The great advantage of such an approach is the inclusion of field evidence, but there are also restrictions, such as the absence of explicit criteria regarding significant effects on the vegetation, and the impossibility to predict future impacts when N deposition changes. Model approaches can account for this. In this paper, we review the possibilities of static and dynamic multispecies models in combination with dynamic soil-vegetation models to (1) predict plant species composition as a function of atmospheric N deposition and (2) calculate critical N loads in relation to a prescribed protection level of the species composition. The similarities between the models are presented, but also several important differences, including the use of different indicators for N and acidity and the prediction of individual plant species vs. plant communities. A summary of the strengths and weaknesses of the various models, including their validation status, is given. Furthermore, examples are given of critical load calculations with the model chains and their comparison with empirical critical N loads. We show that linked biogeochemistry-biodiversity models for N have potential for applications to support European policy to reduce N input, but the definition of damage thresholds for terrestrial biodiversity represents a major challenge. There is also a clear need for further testing and validation of the models against long-term monitoring or long-term experimental data sets and against large-scale survey data. This requires a focused data collection in Europe, combing vegetation descriptions with variables affecting the species diversity, such as soil acidity, nutrient status and water availability. Finally, there is a need for adaptation and upscaling of the models beyond the regions for which dose-response relationships have been parameterized, to make them generally applicable.

Hydrolysed formula is a risk factor for vitamin K deficiency in infants with unrecognised cholestasis.

OBJECTIVES: Vitamin K deficiency (VKD) may cause life-threatening haemorrhages, especially in breast-fed infants with unrecognised cholestasis. Interestingly, hypoallergenic formulas appear overrepresented in reported cases of VKD bleeding (VKDB) in formula-fed infants. We therefore assessed whether the risk of VKD in formula-fed infants with cholestasis is associated with hypoallergenic formulas. PATIENTS AND METHODS: Infants born in the Netherlands between January 1991 and December 2006 with cholestatic jaundice due to biliary atresia (BA) or to α-1-antitrypsin deficiency (A1ATD) were identified in the Netherlands Study Group for Biliary Atresia Registry and the A1ATD registry, respectively. The relative risk (RR) of VKDB in patients with BA or A1ATD was calculated for different formula types. The influence of prior or ongoing breast-feeding on the RR of VKDB was also assessed. RESULTS: A total of 179 infants with either BA (139) or A1ATD (40) were included. One hundred eighteen infants were formula fed; 8 presented with VKD. Six of these 8 infants (75%) received hypoallergenic formula (whey-based hydrolysate in 4). One infant on whey-based hydrolysed formula presented with VKDB. Risk factor analysis revealed that infants receiving hydrolysed, especially whey-based, formula, had a strongly increased risk of VKD (RR 25.0 [6.4-97.2], P < 0.001)) compared with infants receiving regular formula. Prior or ongoing breast-feeding was not significantly associated with VKD. CONCLUSIONS: Infants with cholestasis receiving (whey-based) hydrolysed formula are at increased risk of developing VKD, compared with infants receiving regular formula. Because VKD may lead to serious haemorrhages, infants receiving whey-based hydrolysed formulas may need additional vitamin K supplementation.

Impacts of sampling design and estimation methods on nutrient leaching of intensively monitored forest plots in the Netherlands.

Element fluxes through forest ecosystems are generally based on measurements of concentrations in soil solution at regular time intervals at plot locations sampled in a regular grid. Here we present spatially averaged annual element leaching fluxes in three Dutch forest monitoring plots using a new sampling strategy in which both sampling locations and sampling times are selected by probability sampling. Locations were selected by stratified random sampling with compact geographical blocks of equal surface area as strata. In each sampling round, six composite soil solution samples were collected, consisting of five aliquots, one per stratum. The plot-mean concentration was estimated by linear regression, so that the bias due to one or more strata being not represented in the composite samples is eliminated. The sampling times were selected in such a way that the cumulative precipitation surplus of the time interval between two consecutive sampling times was constant, using an estimated precipitation surplus averaged over the past 30 years. The spatially averaged annual leaching flux was estimated by using the modeled daily water flux as an ancillary variable. An important advantage of the new method is that the uncertainty in the estimated annual leaching fluxes due to spatial and temporal variation and resulting sampling errors can be quantified. Results of this new method were compared with the reference approach in which daily leaching fluxes were calculated by multiplying daily interpolated element concentrations with daily water fluxes and then aggregated to a year. Results show that the annual fluxes calculated with the reference method for the period 2003-2005, including all plots, elements and depths, lies only in 53% of the cases within the range of the average +/-2 times the standard error of the new method. Despite the differences in results, both methods indicate comparable N retention and strong Al mobilization in all plots, with Al leaching being nearly equal to the leaching of SO(4) and NO(3) with fluxes expressed in mol(c) ha(-1) yr(-1). This illustrates that Al release, which is the clearest signal of soil acidification, is mainly due to the external input of SO(4) and NO(3).

Prediction of plant species occurrence as affected by nitrogen deposition and climate change on a European scale.

Plant species occurrence in Europe is affected by changes in nitrogen deposition and climate. Insight into potential future effects of those changes can be derived by a model approach based on field-based empirical evidence on a continental scale. In this paper, we present a newly developed empirical model PROPS, predicting the occurrence probabilities of plant species in response to a combination of climatic factors, nitrogen deposition and soil properties. Parameters included were temperature, precipitation, nitrogen deposition, soil pH and soil C/N ratio. The PROPS model was fitted to plant species occurrence data of about 800,000 European relevés with estimated values for pH and soil C/N ratio and interpolated climate and modelled N deposition data obtained from the Ensemble meteo data set and EMEP model results, respectively. The model was validated on an independent data set. The test of ten species against field data gave an average Pearson's r-value of 0.79. PROPS was applied to a grassland and a heathland site to evaluate the effect of scenarios for nitrogen deposition and climate change on the Habitat Suitability Index (HSI), being the average of the relative probabilities, compared to the maximum probability, of all target species in a habitat. Results for the period 1930-2050 showed that an initial increase and later decrease in nitrogen deposition led to a pronounced decrease in HSI, and with dropping nitrogen deposition to an increase of the HSI. The effect of climate change appeared to be limited, resulting in a slight increase in HSI.

Towards a global-scale soil climate mitigation strategy.

Sustainable soil carbon sequestration practices need to be rapidly scaled up and implemented to contribute to climate change mitigation. We highlight that the major potential for carbon sequestration is in cropland soils, especially those with large yield gaps and/or large historic soil organic carbon losses. The implementation of soil carbon sequestration measures requires a diverse set of options, each adapted to local soil conditions and management opportunities, and accounting for site-specific trade-offs. We propose the establishment of a soil information system containing localised information on soil group, degradation status, crop yield gap, and the associated carbon-sequestration potentials, as well as the provision of incentives and policies to translate management options into region- and soil-specific practices.

The persistent sciatic artery.

BACKGROUND: A persistent sciatic artery (PSA) is a rare vascular anomaly with an estimated incidence of 0.03-0.06%. During early embryonic development, the sciatic artery usually disappears when the superficial femoral artery has developed properly. This study aimed to assess the clinical presentation and outcome of a PSA. METHOD: A systematic review of all cases of PSA published between 1964 and 2007 was performed. RESULTS: In this review, 159 PSAs were described in 122 patients. The mean age at which the PSA was discovered was 57 years, and the incidence was equally distributed with regards to gender. The majority of PSAs was unilateral (70%) and of the complete type (79%). Ninety-one patients (80%) presented with symptoms including intermittent claudication, ischaemia, a pulsating mass or neurological symptoms. An aneurysm was found in 48%, a stenosis in 7%, an occlusion of the PSA in 9% and an occlusion of an artery distal to the PSA in 6% of the subjects. The treatment depended on the symptoms and classification of the PSA. In nine cases (8%), an amputation was required eventually. CONCLUSION: The PSA is a rare anomaly with a high incidence of complications including aneurysm formation and ischaemia that may lead to amputation. Strategies for follow-up could not be deduced from the available literature.

Respiratory muscle strength in stable adolescent and adult patients with cystic fibrosis.

BACKGROUND: Since available studies have provided conflicting results, this study investigated respiratory muscle function and its relationship with exercise capacity, degree of dyspnoea and leg discomfort, and quality of life in patients with Cystic Fibrosis (CF). METHODS: Using a cross-sectional design, 27 clinically stable adolescent and adult patients (f/m: 14/13, age: 26+/-7 years) were included. Data of respiratory muscle strength (P(i)max and P(e)max), lung function (spirometry), peripheral muscle strength (peak isometric quadriceps and hand-grip strength), symptom-limited exercise capacity (modified shuttle test, MST), post-exercise dyspnoea and leg discomfort (Borg scores), and quality of life (CFQ-14+, MRC) were obtained for further analysis. RESULTS: P(i)max of the total patient group was significantly higher than reference values (P(i)max=124+/-32% predicted), and correlated positively with the walk/run distance of the MST (r(s)=0.59, p=0.00). Female patients showed more dyspnoea and a more impaired lung function than male patients. However, P(i)max and P(e)max (% predicted) showed a tendency to be higher in female than in male patients. CONCLUSION: Increased work of breathing will have a conditioning effect on the respiratory muscles, suggesting that training-related inspiratory muscle strength can play a positive role in the limited exercise capacity of CF patients.

Predicting dissolved inorganic nitrogen leaching in European forests using two independent databases.

Regional-scale databases can be particularly useful for identifying relationships between dissolved inorganic nitrogen (N) leaching in forests and environmental drivers, which in turn allow an assessment of the risk of ecosystem damage, such as forest acidification and eutrophication of downstream water bodies. However, detecting the 'signal' of a significant correlate to N leaching against a background of wide variability in other factors requires a large number of sites, and the validation of models developed requires a similarly large number of independent sites. Here we use two large and fully independent databases of forest ecosystems across Europe to develop and validate indicators of N saturation and leaching. One database was used for model development and the other for validating these models. Among 35 variables considered, the most significant indicators of N leaching in the model development database were: the flux of dissolved inorganic N in deposition, mean annual temperature, mean altitude, the site drainage (plot vs catchment), needle- and litter-N concentration, organic horizon C:N ratio, and subsoil pH. Altitude was not a consistent predictor (it was significant in the development database but not in the validation database), and needle and litter N concentration, plot vs catchment, and subsoil pH all showed high intercorrelation with N deposition and so were not significant in models already including N deposition. The most consistent and useful indicators of N leaching were throughfall N deposition, organic horizon C:N ratio and mean annual temperature. Sites receiving low levels of N deposition (<8 kg N ha(-1) y(-1)) showed very low output fluxes of N and were simulated separately from more polluted forests. In general, the models successfully predicted N leaching (mean of +/-5 kg N ha(-1) y(-1) between observed and predicted) from forests at early to intermediate stages of nitrogen saturation but not from nitrogen-saturated sites. Thus, simple relationships developed from combining (1) external drivers (deposition, temperature) and (2) site conditions (nitrogen status of soils) can successfully estimate nitrogen leaching from forests that have not yet been highly damaged by N deposition.

Assessment of the optimal time interval for repeated soil surveys at intensively monitored forest plots.

Statistical methods were developed to assess required changes in the contents and pools of major nutrients and exchangeable base cations in the organic layer and the mineral soil of European forest soils, to derive significant differences. Furthermore a simple element retention model is described and applied to assess the variation time periods, as a function of site and soil characteristics and atmospheric inputs, that are needed before repeating soil surveys in order to assess significant differences in element pools. Time periods that are needed to assess a significant difference have been limited to N in the organic layer and base cations in the mineral layer, since those pools are liable to change caused by nitrogen or acid deposition. Results showed that a time interval of 10 years, which is generally considered for a repetition of the soil survey, might give a significant difference in N and exchangeable base cation pools for approximately 25% and 10% of the plots, only.

Oxygen uptake kinetics in chronic heart failure: clinical and physiological aspects.

One of the hallmark symptoms of patients with chronic heart failure (CHF) is exercise intolerance. Therefore, exercise testing has become an important tool for the evaluation and monitoring of heart failure. Whereas the maximal aerobic capacity (peak VO(2)) is a reliable indicator of the severity and prognosis of heart failure, submaximal exercise parameters may be more closely related to the ability to perform daily activities. As such, oxygen (O(2)) uptake kinetics, describing the rate change of O(2) uptake during onset or recovery of submaximal constant-load exercise (O(2) onset and recovery kinetics, respectively), have been shown to be useful parameters for objectively evaluating the functional capacity of CHF patients. However, their evaluation in this population is not a routine part of daily clinical practice. Possible reasons for this include a lack of standardisation of the assessment methodology and a limited number of studies evaluating the clinical use of O(2) uptake kinetics in CHF patients. In addition, the pathophysiological mechanisms underlying the delay in O(2) uptake kinetics in these patients are not completely understood. This review discusses the current literature on the clinical potency and physiological determinants of O(2) uptake kinetics in CHF patients and provides directions for future research. (Neth Heart J 2009;17:238-44.Neth Heart J 2009;17:238-44.).

M-CSF and IL-34 expression as indicators for growth in sporadic vestibular schwannoma.

Macrophage colony stimulating factor and IL-34 are associated with clinical vestibular schwannoma progression. Investigating the biology behind vestibular schwannoma progression helps understanding tumor growth. Inflammation is important in the microenvironment of neoplasms. Macrophages are major players in the intratumoral infiltrate. These tumor-associated macrophages are known to stimulate angiogenesis and cell growth. M-CSF and IL-34 are cytokines that can regulate tumor-infiltrating macrophages. They are expressed by tumors and form potential targets for therapy. The goal of this study was to investigate these cytokines in vestibular schwannomas and to see if their expression is related to angiogenesis, macrophage numbers, cystic degeneration, and volumetric tumor progression. Immunohistochemical expression of M-CSF and IL-34 was analyzed in ten fast-growing vestibular schwannomas and in ten slow-growing vestibular schwannomas. Expression M-CSF and IL-34 were compared between fast- versus slow-growing and cystic versus non-cystic tumors. Data on macrophage numbers and microvessel density, known from earlier research, was also included. All tumors expressed M-CSF and its expression was higher in fast-growing tumors (p = 0.003) and in cystic tumors (p = 0.035). CD163 expression was higher in tumors with strong M-CSF expression (p = 0.003). All tumors expressed IL-34 as well, but no significant differences were found in relation to clinicopathological characteristics. This study demonstrated the expression of M-CSF and IL-34 in vestibular schwannomas. The results suggest that M-CSF is related to macrophage activity and tumor progression, making it a potential target for therapy. If a similar assumption can be made for IL-34 remains unclear.

Increased virus replication in mammalian cells by blocking intracellular innate defense responses.

The mammalian innate immune system senses viral infection by recognizing viral signatures and activates potent antiviral responses. Besides the interferon (IFN) response, there is accumulating evidence that RNA silencing or RNA interference (RNAi) serves as an antiviral mechanism in mammalian cells. Mammalian viruses encode IFN antagonists to counteract the IFN response in infected cells. A number of IFN antagonists are also capable of blocking RNAi in infected cells and therefore serve as RNA-silencing suppressors. Virus replication in infected cells is restricted by these innate antiviral mechanisms, which may kick in earlier than the viral antagonistic or suppressor protein can accumulate. The yield of virus vaccines and viral gene delivery vectors produced in mammalian producer cells may therefore be suboptimal. To investigate whether blocking of the innate antiviral responses in mammalian cells leads to increased viral vector production, we expressed a number of immunity suppressors derived from plant and mammalian viruses in human cells. We measured that the yield of infectious human immunodeficiency virus-1 particles produced in these cells was increased 5- to 10-fold. In addition, the production of lentiviral and adenoviral vector particles was increased 5- to 10-fold, whereas Sindbis virus particle production was increased approximately 100-fold. These results can be employed for improving the production of viral gene transfer vectors and viral vaccine strains.

Contextual renewal of nicotine seeking in rats and its suppression by the cannabinoid-1 receptor antagonist Rimonabant (SR141716A).

Nicotine-associated paraphernalia such as cigarettes and ashtrays are potent smoking relapse triggers. In addition to these discrete cues, environmental contexts previously associated with smoking elicit strong cigarette craving, indicating that contextual stimuli also contribute to high smoking relapse rates. Nonetheless, little is known about the precise role of these stimuli in smoking relapse and the neuropharmacological mechanisms implicated herein. To address this issue, we determined whether re-exposure to the nicotine self-administration context after long-term extinction reinstates nicotine seeking behavior in rats. Further, we examined the effects of SR141716A (Rimonabant), a selective cannabinoid CB1 receptor antagonist which has been shown to attenuate cue-induced relapse to nicotine seeking, on context-induced reinstatement of nicotine seeking. Rats were trained to self-administer nicotine intravenously (30microg/kg/infusion). Nicotine infusions were paired with an audiovisual compound stimulus. Subsequently, nose poking behavior was extinguished in the presence of this discrete cue in a context different from the self-administration context. Hereafter, rats were injected with 0, 1, or 3mg/kg Rimonabant (i.p.) prior to re-exposure to either the self-administration or the extinction context. Re-exposure to the self-administration context, but not to the extinction context robustly reinstated responding for the discrete nicotine cues, an effect that was dose-dependently attenuated by Rimonabant. This is the first demonstration of contextual renewal of nicotine seeking in rodents after prolonged withdrawal. Further, our results indicate that the endocannabinoid system is involved in context-induced relapse to nicotine seeking, and as such these data provide further evidence for the use of CB1 antagonists in smoking cessation.