Increased sEBR is commonly associated with increased dopamine function based on pharmacological evidence and patient studies. Yet, this hypothesis has not been validated using in vivo measures of dopamine function in humans. To fill this gap, we measured sEBR and striatal dopamine synthesis capacity using [18F]DOPA PET in 20 participants . Our results, based on frequentist and Bayesian statistics, as well as region‐of‐interest and voxel‐wise analyses, argue against a positive relationship between sEBR and striatal dopamine synthesis capacity. They show that, if anything, the evidence is in favour of a negative relationship. These results, which complement findings from a recent study that failed to observe a relationship between sEBR and dopamine D2 receptor availability, suggest that caution and nuance are warranted when interpreting sEBR in terms of a proxy measure of striatal dopamine.Dopamine is central to a number of cognitive functions and brain disorders. Given the cost of neurochemical imaging in humans, behavioural proxy measures of dopamine have gained in popularity in the past decade, such as spontaneous eye blink rate ( In turn, striatal dopamine dysregulation is a common feature across many psychiatric disorders in which those functions are impaired, including addiction, schizophrenia, attention deficit hyperactivity disorder and depression  receptor availability. Only two PET studies to date have tackled this question, and both of these focused on dopamine receptor availability. The first study by Groman et al.  receptor availability was positively correlated with sEBR, as well as with D2 receptor agonist‐induced changes in sEBR. However, a recent study by Dang et al.  and 15 matched healthy controls (HCs). The PET data from these participants have been reported in a previous study comparing baseline dopamine synthesis capacity between groups .Nine participants  were not included due to technical problems with the electro‐oculography (EOG) recordings . Data loss was related to EOG amplifier saturation (output voltage of the amplifier exceeding its operating range sensitivity). Three participants were kept in the analyses despite minor data loss . One gambler was further excluded because he met the DSM‐IV‐TR criteria for past year marijuana dependence, which is known to influence sEBR measures .18F]DOPA and increase tracer availability in the brain while having no psychotropic side effects. The participants further performed computerized tasks not reported here.The study took place at the Radboud University Medical Center. Upon arrival of the participants, spontaneous eye blink rate (sEBR) was measured using EOG recordings (see detailed description below). Approximately 1 h before entering the PET scanner, participants received 150 mg of carbidopa and 400 mg of entacapone to reduce peripheral metabolism of DOPA into an antecubital vein. Images were reconstructed with an ordered subset expectation maximization algorithm with weighted attenuation and time‐of‐flight recovery, scatter corrected and smoothed with a 4‐mm FWHM kernel.All PET scans were acquired at the department of Nuclear Medicine of the Radboud University Medical Center using a Siemens mCT PET/CT camera . Participants were positioned as comfortably as possible, in a supine position, with the head slightly fixated in a headrest to avoid movement. First, a low‐dose CT was made for attenuation correction of the PET images, followed by an 89‐min dynamic PET scan. The scan started at the same time as the bolus injection of the [3) was obtained using a 3 Tesla MR Siemens scanner at the Donders Centre for Cognitive Neuroimaging and used for co‐registration with the PET data.A high‐resolution anatomical scan  frame to correct for movement during scanning using SPM8 (http://www.fil.ion.ucl.ac.uk/spm/). The mean [18F]DOPA image and the realigned frames were coregistered to the structural MRI scan using SPM8. Higher [18F]DOPA uptake, indexed by higher Ki values, is established to reflect higher dopamine synthesis capacity . Specifically, we retained the template voxels in which Ki values were at least three standard deviations above the template mean, thus approximating the anatomical boundaries of the striatum , we used rank correlation coefficients (Spearman ρ). We also performed partial regression analyses, in which beta coefficients were estimated using bootstrapping in spss. Bayesian analyses were conducted using jasp .In contrast to our prediction, frequentist statistics indicated a negative relationship between sEBR and Ki values , indicating that our data are 10 times more likely under the null hypothesis of no positive relationship than under the alternative hypothesis of a positive relationship. Importantly, a Bayes factor robustness check showed that this conclusion holds even when using strong prior beliefs in the existence of a positive correlation  in favour of a negative relationship between sEBR and Ki values (H1 vs. H0 defined as an absence of negative relationship).Given the unexpectedness of this finding, we sought to quantify the evidence for the null hypothesis that sEBR and Ki values are not positively correlated (H0), as opposed to positively correlated (H1). We used a Bayesian analysis with an uninformative (flat) prior distribution over the  interval. The relative evidence in favour of H0 over H1 was strong , while the Bayesian correlation analysis indicated moderate to strong evidence against hypothesis H1 of a positive correlation (BF01 = 9.95). When splitting the data as a function of gambling status, we observed a negative association between sEBR and Ki values in both groups . Although P‐values did not reach significance, Bayesian analyses similarly revealed moderate evidence against a positive correlation , although, of course, these values should be interpreted with caution given the small sample sizes of these subgroups.To assess the robustness of our results, we performed a number of control analyses. First, we re‐ran our analyses after excluding the data from the three participants with incomplete sEBR datasets. The results remained unchanged, showing a negative correlation between sEBR and striatal dopamine syntheses , while corresponding Bayes factors again provided strong evidence in favour of the null hypothesis H0 .Finally, we verified that our results were not influenced by the method used to define eye blinks. Indeed, in our main analysis, we used a relatively low detection threshold of 100 μV, which has the advantage of capturing most blinks (high sensitivity), but has the disadvantage of increasing the likelihood of false positives due to muscular artefacts (decreased specificity), which then requires manual (i.e. subjective) editing. To circumvent that issue, we re‐ran our analyses using more conservative thresholds (200 and 300 μV), which provide increased specificity and thus eliminate the need for manual editing (at the cost of decreased sensitivity). The resulting sEBR measures were again negatively correlated with Ki values, although not significantly  of the dopamine system correlate(s) with sEBR. Here we tested the hypothesis of a positive relationship between sEBR and striatal dopamine synthesis capacity, based on the proposal that sEBR is positively related to striatal dopamine function , as this would predict a positive relationship between sEBR and dopamine synthesis capacity, in contrast to our results. These inconsistencies certainly call for further research to better elucidate the dopaminergic mechanisms underlying sEBR.Importantly, our data speak specifically to dopamine synthesis capacity and do not preclude correlations of sEBR with other aspects of the dopamine system, including (changes in) dopamine D2 receptor availability. It has been suggested that sEBR might primarily reflect striatal D2 receptor activity, based on the observation of a positive correlation with D2 receptor availability in monkeys (Groman n = 20) is relatively small, although one should note that it is larger than the samples used in many of the psychopharmacological studies that have investigated dopaminergic drug effects on sEBR . In addition, as argued by Dang et al. (This study is not devoid of limitations. First, our sample size (, et al.  between g et al. , the useet al., Another aspect that may be perceived as a limitation is the use of a mixed population of healthy participants and pathological gamblers. While we acknowledge that pathological gamblers are not typical individuals and are characterized, among other things, by elevated striatal dopamine synthesis (van Holst To conclude, our study does not support the hypothesis of a positive relationship between sEBR and striatal dopamine synthesis, and if anything, provides evidence against it. Even though it is based on a modest sample size and needs to be replicated in a larger sample – which we are currently attempting to do, it warrants caution for future studies that may be tempted to use sEBR as a proxy measure of striatal dopamine synthesis capacity.WJJ serves as a consultant to Genentech, Novartis, and Bioclinica. All other authors report no biomedical financial interests or potential conflicts of interest.https://doi.org/10.6084/m9.figshare.5878777.The study data and analysis scripts are available online at GS, RJvH and RC designed the study. RJvH and LKJ collected the data. GS, RL, RJvH, FdB, MJ, ASB and WJJ performed data analysis. GS and RL wrote the first draft of the manuscript. RJvH and RC edited the manuscript. We wish to thank Heleen Slagter for kindly sharing her analysis scripts for sEBR data.EOGElectro‐oculographyMRIMagnetic Resonance ImagingPETPositron Emission TomographysEBRspontaneaous Eye Blink Rate Click here for additional data file.


Mycobacterium bovis, which belongs to the Mycobacterium tuberculosis complex. Mycobacterium bovis have been described to be responsible of most cases of bovine tuberculosis. Although M. tuberculosis, M. africanum and non-complex mycobacteria were isolated from cattle.Bovine tuberculosis (BTB) is caused by M. bovis in Morocco.In Morocco, so far, no molecular studies were conducted to characterize the strains responsible of BTB. The present study aims to characterize The present study was conducted in slaughterhouses in Rabat and El Jadida. Samples were collected from 327 slaughtered animals with visible lesions suggesting BTB.n = 215) of them were acid-fast (AF). Sixty eight per cent of the AF positive samples were confirmed as tuberculous mycobacteria (n = 147), 99% of these (n = 146) having RD9 and among the latter, 98% (n = 143) positive while 2% (n = 3) negative for RD4A total of 225 isolates yielded cultures, 95% . The most prevalent spoligotypes were SB0121, SB0265, and SB0120, and were already identified in many other countries, such as Algeria, Spain, Tunisia, the United States and Argentina.M. bovis spoligotypes. On the other hand, the desert of Morocco could be considered as an efficient barrier preventing the introduction of BTB to Morocco from West Central and East Africa. Our findings suggest a low level endemic transmission of BTB similar to other African countries. However, more research is needed for further knowledge about the transmission patterns of BTB in Morocco.The shared borders between Algeria and Morocco, in addition to the previous importation of cattle from Europe and the US could explain the similarities found in  Mycobacterium bovis, which belongs to the Mycobacterium tuberculosis complex (MTBC). M. bovis has the particular ability to infect a wide range of host species other than cattle  and humans, [M. bovis [Meles meles), the Possum (Trichosurus vulpecula) [Bovine tuberculosis (BTB) is a chronic granulomatous caseous-necrotising inflammatory disease that mainly affects the lungs and their draining lymph nodes, it can also affect other organs, depending on the route of infection –3. BTB i humans, , howeverM. bovis . There elpecula) .M. bovis can be transmitted between animals by inhalation of aerosols and the ingestion of contaminated food. The transmission is enhanced by many risk factors, mostly related to intensive livestock system [M. bovis to humans occurs by consumption of unpasteurized infected raw milk and by the contact with infected cattle [k system . The trad cattle .While BTB is still occurring in some developed countries in low prevalence –10 becauIn Morocco, the agriculture sector is of key importance to the economy, representing approximately 14% of total gross domestic product (GDP) (75bn MAD/ €6.6bn) and approximately 7% of exports 2009). Livestock represents 38% of the total agriculture sector GDP . Both ex09. LivesOfficially, skin test and slaughter is the current control strategy applied in Morocco; however this strategy is not fully applied as it is not respected; in addition, there is no systematic BTB screening of cattle at a national level .M. bovis among human TB patients [M. bovis on certain groups of the population, and report a potential underestimation of the prevalence of zoonotic human TB [In a review published by Muller et al. in 2013, the proportion of zoonotic human tuberculosis (TB) among all TB cases was estimated at 1.4% for the non-African countries, and at 2.8% in Africa . The higpatients . Even ifhuman TB , 25.M. bovis is the most recent strain in his lineage showing the deletion of RD4 [The phylogeny of MTBC showed recently that the strains found in animals belong to a single lineage which showed the deletion of the “Region of Difference” 9 (RD9) , 26. Indn of RD4 .M. bovis from the other strains of the MTBC [Molecular deletion typing had been found to be an important tool to differentiate the MTBC . Patternthe MTBC –30.Currently there is no data available in Morocco about the molecular characterization of BTB and the prevalence of MTBC among slaughtered cattle. The aim of the present study was to characterize the strains of MTBC which are responsible for BTB among the slaughtered cattle in Morocco.The study was conducted in two slaughterhouses in Morocco, one in Rabat and another in EL Jadida, which are two coastal cities separated by 200 km. The cattle slaughtered in these two slaughterhouses come from the rural areas surrounding the two cities and also from many other areas of the country. Individual information of every animal, such as gender, age, breed, and the possible origin were recorded in our database, as well as the date of sampling. The sample collection was performed in Rabat from March to July 2015 and in El Jadida from June 2014 to April 2015.The samples were conveyed to the Veterinary and Agriculture Institute (IAV) in Rabat and stored in −20 °C until their treatment.Prior to treatment, samples were thawed overnight at 4 °C. Subsequently, samples were desiccated to remove adipose tissue, and 5 g of the desiccated lesions were mixed with sterilized sand and 10 ml of phenol red. The solution (7.5 ml) was placed in a 15 ml conic tube, 2.5 ml of NaOH 1 N was added at room temperature for 10 min, and then HCl 6 N was added for sample neutralization. As a final step, the tube was centrifuged for 25 min at 3500 rpm.The supernatant was discarded and pellet was distributed in two of the four pre-tested culture media: Lowenstein-Jensen (LJ), LJ with glycerol (LJG) or pyruvate (LJP) and Herrold according to the availability in the laboratory. Cultures were incubated for 12 weeks at 37 °C and observed daily for growing colonies during the first week then weekly from the second week onwards.All the grown cultures were deactivated by adding a loopful of mycobacterium colonies to 1 ml of sterilized water contained in small tubes. The samples were then inactivated for 1 h at 90 °C.Mycobacterium molecular characterization was performed using Multiplex polymerase chain reaction (PCR). The PCR was performed in the TB laboratory at Swiss TPH. We performed first MrpoB PCR in order to differentiate MTC and NTM as described earlier [M. bovis and M. tuberculosis from other species of the MTBC by assessing the presence or absence of Regions of Difference 9 and 4 (RD). The analysis was carried out as previously described [ earlier . Deletioescribed , 28.Mycobacterium bovis molecular typing database [Spoligotyping was performed as previously described . Spoligodatabase . All then = 215) of the total sampled animals were analysed by Ziehl-Neelsen(ZN), 68% (n = 147) of the latter were ZN positive and heat-killed for further molecular typing.In the present study, a total number of 8658 animals were examined. Three hundred and twenty seven animal presented gross visible lesions (3.7%) and were cultured, 66%  of the 215 samples which were AF positive were confirmed as MTBC strains using MrpoB PCR. Thirty two per cent (n = 69) of animals were negative for MrpoB.A total of 68%  of the samples were confirmed to be M. bovis as they were positive to the RD4 pcr. Three samples were negative for RD4.The samples confirmed as MTBC strains were all positive to RD9 deletion typing and then confirmed to be not M. bovis samples, 30% were female while 70% were male. The predominant breed in the positive animals was Crossbreed prime Holstein.Out of the total of the confirmed n = 33). 16.9% (n = 23), 9.6% (n = 13) respectively. They were shown to belong to BOV-1 family. Two other spoligotypes were found in 9 and 6 samples respectively, SB0125 and SB0869. Three spoligotype patterns had no SIT reference on the SITVIT database and were designated as orphan. Ten isolates presented nine undescribed spoligotypes, which were submitted to M. bovis website (www.mbovis.org)  was low compared to the prevalence found previously using tuberculin skin test in the last national survey performed in 2004 [Surprisingly, no non-tuberculous mycobacteria was found, while in the African context, among slaughterhouse gross visible lesions, 3.3% in Mali , and 9% M. bovis, three samples were negative to RD4. While two spoligotypes were typical for the caprae family, the third sample was a new spoligotype. Studies in other Countries  found M. tuberculosis in cattle [Almost 52.4% of the overall sample size was confirmed to be n cattle , whereasM. bovis and/or that there is probably little trade of cattle between Morocco and Mali [The most predominant spoligotype pattern found in Morocco is SB0121, belonging to the family BOV 1, this spoligopattern was already reported in Algeria , in Tuniand Mali .M. caprae family, of which one was found as well in M.bovis database as SB0866, this spoligotype was already reported in Spain from one goat, one cattle and one pig in 2011 [Two spoligotypes of three samples belonged to the  in 2011 .The similarities found in some spoligopatterns between Morocco and Algeria could be explained by the shared borders between the two countries, in addition some patterns found in Morocco were previously reported in the United States and in Argentina, two countries from where Morocco have previously imported cattle.Spoligotypes of African 1 and African 2 clonal complexes were not found among our characterized isolates , 52. AfrM. bovis infection in two Moroccan abattoirs has to be interpreted with caution. Firstly, the abattoir prevalence reflects more the prevalence in young bulls and old cows rather than the whole population. Secondly, not all granuloma yielded a bacteriological isolate, hence the true prevalence may be much higher than the one observed. Overall, our findings reflect rather the epidemiological situation of a low level endemic transmission similar to other African countries rather than the one of a peri-urban intensive system [The relatively low prevalence of proven e system , 54. More system .M. bovis represented a high amount of granulomatous lesions detected in the abattoirs of Rabat and El Jadida. Spoligotype suggests a link of Moroccan BTB to Europe, rather than to Africa, highlighting then the potential of the Moroccan desert barrier for BTB introduction to Morocco from sub-Saharan Africa. Further investigations of BTB strains using new molecular techniques such as whole genome sequencing are needed to clarify more the potential links between Moroccan BTB strains and those of Europe and other African countries.This study presents the first molecular characterization of BTB isolates from Moroccan cattle.


However, the low conductivity of ions and bulk electrons hinder its rapid development in lithium-ion batteries (LIB). In order to improve the electrochemical performances of TiO2 nanomaterials as anode for LIB, hierarchically porous TiO2 nanofibers with different tetrabutyl titanate (TBT)/paraffin oil ratios were prepared as anode for LIB via a versatile single-nozzle microemulsion electrospinning (ME-ES) method followed by calcining. The experimental results indicated that TiO2 nanofibers with the higher TBT/paraffin oil ratio demonstrated more axially aligned channels and a larger specific surface area. Furthermore, they presented superior lithium-ion storage properties in terms of specific capacity, rate capability and cycling performance compared with solid TiO2 nanofibers for LIB. The initial discharge and charge capacity of porous TiO2 nanofibers with a TBT/paraffin oil ratio of 2.25 reached up to 634.72 and 390.42 mAh·g−1, thus resulting in a coulombic efficiency of 61.51%; and the discharge capacity maintained 264.56 mAh·g−1 after 100 cycles, which was much higher than that of solid TiO2 nanofibers. TiO2 nanofibers with TBT/paraffin oil ratio of 2.25 still obtained a high reversible capacity of 204.53 mAh·g−1 when current density returned back to 40 mA·g−1 after 60 cycles at increasing stepwise current density from 40 mA·g−1 to 800 mA·g−1. Herein, hierarchically porous TiO2 nanofibers have the potential to be applied as anode for lithium-ion batteries in practical applications.Titanium dioxide (TiO The increasing demand for energy has heightened the need for the development of renewable resources because fossil fuels, which are currently the most common energy resources, are a finite resource –4. Batte2) is considered to be an alternative anode material to graphite, which can be attributed to the superior advantages of titanium dioxide such as low-cost, eco-friendliness, nontoxicity and high abundance [2 matrix at higher voltages (at least 1.5 V vs Li/Li+) [2 possesses a high theoretical specific capacity of 335 mAh·g−1 [So far, among all the commercial lithium-ion batteries, graphite plays an extremely important role in anode materials; nevertheless, structural deformation, electrical disconnection and the initial loss of capacity hinder its further development –10. Tita Li/Li+) –13. TiO2 mAh·g−1 –16. Howe2 is to prepare nanostructures [One effective method to improve the electrochemical performance of TiOructures –20 that ructures –22. Elecructures –25.2, SiO2, ZrO2, SnO2, V2O5, GeO2 and Al2O3. A microemulsion system was composed of a metal alkoxide and an oil phase as, respectively, continuous phase and dispersion phase in the system [2 nanorods via ME-ES and the application as photoanode material for dye-sensitized solar cells [2/TiO2 composite nanofibers were prepared successfully by ME-ES and subsequent calcination [Core–shell nanofibers are normally prepared by coaxial electrospinning. However, coaxial electrospinning limits the further development of core–shell nanofibers because of its complicated spinneret, the limited number of suitable fluids and the unstable structure of the fibers. Recently the method of ME-ES has been developed to fabricate hierarchical nanofibers without the need of a complex spinneret. It is a simple and versatile way to prepare nanofibers with hollow or multichannel structures . More ime system . The mice system . Chen etar cells . Accordicination .2 nanofibers as anode for LIB. Cho et al. prepared fiber-in-tube TiO2 nanofibers as anode material for LIB and the initial discharge and charge capacity were 231 and 170 mAh·g−1, respectively [2 nanofibers were fabricated by Tang et al. and demonstrated excellent rate capability and cycling performances when used as anode in LIB [2 nanofibers fabricated via ME-ES used as anode material for LIB. So in this work, the ME-ES solution was firstly electrospun to prepare nanofibers, then as-spun nanofibers were calcined at 500 °C to form porous TiO2 nanofibers. The electrochemical performances of hierarchically porous TiO2 nanofibers as anode material for LIB were investigated in detail.There are various reports about porous TiOectively . Furthere in LIB . HoweverMw = 1300000) was purchased from Tianjin Bodi Chemical Reagent Co., Ltd. . All chemicals were used as received without further purification.Tetrabutyl titanate (TBT), paraffin oil, absolute ethanol, acetic acid and cetyltrimethylammonium bromide (CTAB) were supplied by Shanghai Chemical Regents Co. . Additionally, polyvinyl pyrrolidone (PVP)  of 2.25, 1.9 and 1.55 were added slowly under stirring for 12 h to obtain a microemulsion.−1. The as-spun nanofibers with TBT/paraffin oil ratios of 2.25, 1.9 and 1.55 were marked as sample A1, sample B1 and sample C1, respectively.The prepared microemulsion was filled in a 30 mL syringe with a blunt-end stainless steel needle, which had an inside diameter of 0.3 mm. Then the microemulsion was electrospun into nanofibers by a custom-made electrospinning setup consisting of a high voltage power supply, a syringe pump and a grounded nanofiber collector covered with aluminium foil. During electrospinning, a positive voltage of 20 kV was applied between the needle tip and grounded collector at a distance of 15 cm. The flow rate of the spinning solution was maintained at 2 mL·h2 nanofibers. The porous TiO2 nanofibers obtained after calcination with TBT/paraffin oil ratios of 2.25, 1.9 and 1.55 were named as sample A2, sample B2 and sample C2, respectively. In other words, sample A2 corresponded to the calcined sample A1, sample B2 and sample C2 were the same naming method as sample A2.Subsequently, the as-spun nanofibrous mats were calcined in a quartz tube furnace in air atmosphere at 500 °C for 2 h to yield hierarchically porous TiO2 nanofibers in air atmosphere with a heating rate of 10 °C/min. X-ray diffraction patterns of porous TiO2 nanofibers were recorded on a Bruker D8 Advance X-ray diffractometer with Cu Kα radiation (wavelength λ = 1.54 Å) at a scanning speed of 4 °C/min. A Hitachi field emission scanning electron microscope  was employed to observe the surface morphology of porous TiO2 nanofibers. Prior to the FE-SEM examination, all the specimens were sputter-coated with gold to avoid charge accumulations. Transmission electron microscopy was conducted on a JEOL JEM-2100 transmission electron microscopy unit at an accelerating voltage of 120 kV. The specific surface area and pore structure of porous TiO2 nanofibers were characterized with a physisorption analyzer .Thermogravimetric analysis (1100SF) was employed to characterize the thermal decomposition of as-spun TiO2 nanofibers (60 wt %), carbon black (20 wt %) and poly(tetrafluoroethylene) (20 wt %) to form a homogeneous paste. Subsequently, the paste was coated uniformly on the nickel net and then dried in a vacuum oven at 60 °C for 12 h. Prior to cell assembly, the electrodes were cut into circular disks with a diameter of 16 mm. The testing cells employed Celgard 2300 membrane as separator, lithium foil as both reference and counter electrode, and 1 M LiPF6 dissolved in ethylene carbonate (EC), dimethyl carbonate (DMC) and ethylene methyl carbonate (EMC)  as the electrolyte, respectively. Coin cells were assembled in an argon-filled glove box. The galvanostatic discharge–charge tests were conducted at the constant current density of 40 mA·g−1 in the voltage range of 0.01–3.0 V (vs Li+/Li). The cyclic voltammetry curves were tested at a scanning rate of 0.1 mV/s over the voltage range of 0.01–3.0 V (vs Li+/Li) with a CHI660e electrochemical workstation.The electrodes were prepared by mixing thoroughly porous TiO2 nanofibers is presented in The fabrication process of hierarchically porous TiO1 are shown in 2 nanofibers. 2, sample B2 and sample C2. It can be observed that after calcination, the nanofibers had the length of a few micrometers and a diameter of hundreds of nanometers. Moreover, from Thermogravimetry (TG) and differential thermogravimetry (DTG) curves of as-spun sample A2 calcined at 500 °C for 2 h in air atmosphere. All the peaks can be indexed to anatase TiO2 . The diffraction peaks at 2θ = 25.281, 37.800, 48.049, 53.890, 55.060, 62.688, 70.309 and 75.029° correspond to the (101), (004), (200), (105), (211), (204), (220) and (215) reflections, respectively. It is notable that there was no peak of any other phase in 2 nanofibers. Considering that all the porous TiO2 nanofibers were calcined at 500 °C, all of them should be expected to be anatase TiO2. More importantly, among the various polymorphs of TiO2, the anatase polymorph possesses an open crystal structure resulting from the stacking of zigzag units, which would benefit the intercalation/deintercalation of Li+ ions.1 (a), sample B1 (c), and sample C1 (e) as well as cross-sectional SEM images of sample A2 (b), sample B2 (d), sample C2 (f) and solid TiO2 nanofibers (g), and representative TEM images of sample A2. In addition, the inset SEM images are the corresponding images with the higher magnification. The as-spun nanofibers have a smooth surface and relatively uniform distribution. Nevertheless, it can be observed that with decreasing relative content of TBT/paraffin oil, the surface morphology looks worse, and the distribution of the fiber diameter becomes increasingly non-uniform. This is because the butoxyl groups in TBT serve as additional surfactant, which is advantageous to form the stable interfaces between the two phases, thus forming a stable microemulsion. In other words, the decreased content of TBT caused increasing instability of the microemulsion, so that the morphology of the nanofibers became worse. In addition, the most significant phenomenon is that the number of channels in each nanofiber decreased as the content of paraffin oil increased. This is because the pores stemmed from the vacancies of oil droplets after calcination, and a larger oil droplets were formed when more paraffin oil was added into the microemulsion [2 nanofibers in emulsion . The SEM2, sample B2, sample C2 and solid TiO2 nanofibers are shown in 2, sample B2, sample C2 and solid TiO2 nanofibers were approximately 69.9, 48.2, 43.1 and 35.6 m2/g, respectively. It is noteworthy that the specific surface area of sample A2 was almost two times that of solid TiO2 nanofibers, which demonstrated the fact that ME-ES was indeed an effective method to enlarge the specific surface area. The specific surface area plays a very important role in the performance of electrodes for lithium-ion batteries. The merits of porous nanofibers with a higher specific surface area lie in the higher lithium-ion flux across the interfaces and the larger contact area between the electrode and electrolyte [2 should have the best performances as the electrode of lithium-ion battery in theory. In addition, 2, sample B2 and sample C2. According to the analysis of pore-size distribution with the Barrett–Joyner–Halenda (BJH) method, the average pore size increased gradually, and the average pore diameter was 17.8, 18.5 and 20.1 nm for sample A2, sample B2, sample C2, respectively. Moreover, it can be observed clearly that the average diameter of macropores increased with the ratio of TBT/paraffin oil decreasing. The results confirmed the presence of mesopores and macropores in the porous TiO2 nanofibers.Nitrogen adsorption–desorption isotherms of sample Actrolyte –35. Here2 nanofiber electrodes were investigated by cyclic voltammetry (CV) test using lithium foil as counter electrode and reference electrode. The CV curves of sample A2 and solid TiO2 nanofibers examined in the range of 0.01–3 V (vs Li+/Li) at a scanning rate of 0.1 mV/s are shown in Figure S1 , sample B2 (b) and sample C2 (c) at a current density of 40 mA·g−1. It is apparent that all the charge or discharge profiles were similar, thus implying that the processes of lithium insertion or extraction were the same. There were obvious voltage plateaus in all discharge curves at around 1.72 V and charge curves at around 1.93 V, which were due to the phase transition between tetragonal anatase and orthorhombic Li0.5TiO2 [2, sample B2 and sample C2 were 634.72, 583.44 and 522.65 mAh·g−1, and the corresponding charge capacities were 390.42, 367.85 and 279.82 mAh·g−1, resulting in coulombic efficiencies of 61.51%, 63.05% and 53.54%, respectively. It was worth noting that the specific capacities increased accordingly as the specific surface area of the samples increased, which further confirmed that specific surface area plays a vital role in the performance of the LIB anode material. Sample A2 with a specific surface area of 69.9 m2·g−1 exhibited the best performance among all the samples, and was chosen for further investigation.i0.5TiO2 . The ini2 and solid TiO2 nanofibers. As can be seen from + into TiO2 nanofibers [2 and solid TiO2 nanofibers were 264.56 (261.61) and 198.96 (198.12) mAh·g−1, respectively. It is apparent that the cycling performance of sample A2 shows significant improvement compared with solid TiO2 nanofibers, which can be attributed to the larger specific surface area of sample A2. As mentioned in 2 was 61.51% for the first cycle, whereas the coulombic efficiency of solid TiO2 nanofibers was 57.21%. And the coulombic efficiency remained relatively constant in the following cycles due to the relatively stable cycling performance. The rate capability plays a crucial role in the high-power applications of LIB, therefore rate capability of both samples was evaluated by stepwise increasing the current density. The specific capacity decreased gradually with increasing current density. The electrode delivered reversible discharge capacities of 315.14, 166.47, 142.55, 117.56, 96.42, 50.195 mAh·g−1 at the current density of 40, 80, 160, 320, 400, 800 mA·g−1, respectively. Despite the fact that the capacity could not recovered fully, the corresponding specific capacity of sample A2 was as high as 204.53 mAh·g−1 when the current density returned back to 40 mA·g−1, which was much higher than the values of solid TiO2 nanofibers. Sample A2 as anode for LIB demonstrated excellent rate capability, which stemmed mostly from the hierarchically porous structure and the large specific surface area.nofibers . The cap2 and solid TiO2 nanofibers. Nyquist impedance plots obtained for sample A2 and solid TiO2 nanofibers in a fresh cell are presented in Figure S2 (Rct) and the straight line in the low-frequency region represents the diffusion and accumulation process of lithium ions in the electrode [2 was about 235 Ω, whereas the charge–transfer resistance of solid TiO2 nanofibers was about 425 Ω. Herein, the lower charge–transfer resistance of sample A2 can be ascribed to the hierarchically porous structures of sample A2 facilitating charge transfer at the electrolyte–electrode interfaces [2 with other reference results.Electrochemical impedance spectroscopy (EIS) was carried out on the electrode of sample Aigure S2 . The plolectrode ,29,39. Aterfaces –42 (Figu2 nanofibers were fabricated successfully via single-nozzle microemulsion electrospinning. Moreover, multichannel TiO2 nanofibers displayed excellent lithium-ion storage properties compared with normal solid TiO2 nanofibers when used as anode for LIB. The initial discharge and charge capacities of porous TiO2 nanofibers with a TBT/paraffin oil ratio of 2.25 reached up to 634.72 and 390.42 mAh·g−1; and the discharge capacity was 264.56 mAh·g−1 after 100 cycles. In addition, TiO2 nanofibers with a TBT/paraffin oil ratio of 2.25 still obtained a high reversible capacity of 204.53 mAh·g−1 when the current density returned back to 40 mA·g-1. The results confirmed microemulsion electrospinning is indeed a simple and versatile method to prepare porous nanofibers with large specific surface area and the prepared nanofibers improve greatly their electrochemical performance in practical applications.Hierarchically porous TiOThe Supporting Information presents CV curves and Nyquist impedance plots.File 1Additional experimental data.


Function is like beauty—its definition lies in the eye of the beholder or, in this case, the researcher. At the broadest level, we define organismal function—the function that the protein plays in the overall organism. This function can be observed by understanding the impact on the organism of deletion or mutation of the protein. Physiological function is the function the protein plays in pathways, such as metabolic or signaling pathways. Another level of function is the cellular level. Approaches to understanding cellular function attempt to identify a protein’s interaction partners and its location within the cell. Biochemical or molecular function is another level of function, one that identifies the molecular functional details of a functional site, including reaction mechanism, substrate binding, and molecular details of the binding of regulatory molecules. The International Union of Pure and Applied Chemistry (IUPAC) Enzyme Classification (EC) system was an early approach to identifying molecular function. The Gene Ontology (GO) system of classifying function recognizes ways of defining function, using distinct cellular components, molecular function, and biological process hierarchies . The empWith the ongoing genome sequencing projects, the number of predicted proteins continues to increase exponentially. Understanding the biological role of these protein sequences requires an understanding of their functions, a goal that has resulted in many experimental and computational methods to evaluate gene product function. Large-scale experiments have aided in identification of protein-protein interactions, nonprotein binding partners, and expression levels, each of which define some manner of cellular function of the gene product. Other large-scale projects, such as the Enzyme Function Initiative  have foComputational methods also abound. Our goal here is not to provide a thorough review but rather to provide a brief overview of new work in the field, with illustrative examples. The most commonly used approach is function annotation transfer, exemplified by common application of BLAST and Position-Specific Iterative BLAST (PSI-BLAST) ,4. In itOther approaches to understanding molecular function involve motifs, either sequence motifs exemplified by PRINTS  and PROSFinally, several groups have developed methods to cluster proteins into functionally relevant groups. These approaches—exemplified by Genome Modelling and Model Annotation (GEMMA) , SubfamiThe emphasis of this Focus Feature is molecular function, particularly for enzymes. A primary goal of molecular function analysis is to identify and understand the role of mechanistic or functional determinants, patterns of amino acids that distinguish one functional group from another. These are typically the amino acids that enable substrate binding and catalytic mechanism. Such features distinguish one molecular functional family from another. Their identification has long been the subject of expert analysis in the pharmaceutical and structure-based drug discovery programs, as these features are used to design the most specific inhibitors for a given functional site.Understanding molecular functional determinants would also provide a better understanding of evolution. Contemporary protein superfamilies are the result of numerous genetic events, including gene duplications and horizontal transfers. The relationship between molecular function and evolution has long been debated . Indeed,Hierarchy is inherent in molecular function. Early on, this hierarchy was recognized in the IUPAC EC system, in which each enzyme was assigned a 4-digit EC number, W.X.Y.Z. In the EC system, wherein W represents 1 of 6 main types of chemical reactions (classes) to which the enzyme belongs, X indicates a more detailed level of reaction, i.e., subclass, Y indicates the sub-subclass , and Z is the serial number of the enzyme in its sub-subclass and describes substrate specificity, including pertinent cofactors.Hierarchy is also represented in the GO system of molecular function classification . As an eThe Structure-Function Linkage Database (SFLD) also defines a molecular functional hierarchy, in this case as superfamily, subgroup, and family  26]. Su. Su26]. A key feature of annotation transfer methods is that the level of functional hierarchy must correlate with the similarity method and mathematical and statistical comparison that is being done . An illuThe goal of this Focus Feature is to present some recent work on determining molecular function and the details of features that distinguish one molecular function from another in a superfamily. Three research groups take the approach of clustering proteins into functionally relevant groups. The fourth research group describes a computational approach to enumerating potential reaction products in a protein superfamily, i.e., an approach to identifying what each of those functionally relevant clusters within a superfamily might do.The contribution by Orengo and her colleagues describes their most recent work on clustering the beta lactamase protein superfamily . BuildinThe contribution of de Melo-Minardi and her colleagues  describeA third method, MISST , for cluNotably, the above methods are not function prediction methods because, once functionally relevant clusters are identified, the key question remains to be answered: What do the proteins in each cluster actually do? That is, what is the reaction mechanism of the proteins in each cluster, and what are the substrates utilized and products produced by each cluster? Jacobson and his colleagues present an approach to answering that question, which builds on previous work on the triterpenoid synthases that utilized docking to identify potential substrates and functional families . Their cA common feature of several of the clustering approaches in this Focus Feature is the definition of a “functional site signature.” At its simplest, a signature consists of the “mechanistic determinants” or “specificity-determining positions” or “functional determinants”—only those residues that are actively involved in substrate binding or catalytic mechanism  by Fetrow and colleagues  and an AA key challenge for clustering proteins in functionally relevant ways is the identification of the number of clusters that are optimal for defining all distinct molecular functions within a protein superfamily—that is, the number of clusters that captures biologically relevant functional boundaries that correlate with the level of molecular functional hierarchy being pursued . The preAnother key challenge for methods aimed at molecular function identification is validation. Isofunctional clusters, defined as a family by SFLD, have been defined and experimentally validated for only a small number of families. For example, the enolase superfamily and its functionally relevant families have been studied extensively –41, and Clearly, our understanding of molecular function continues to evolve. Each of the methods described in this Focus Feature builds on previous work, and each continues to aid our understanding of molecular function and relationships between functionally relevant clusters within a protein superfamily. For the future, much new work seeks to improve automated and “agnostic” approaches for functional inference, while other communities, such as Critical Assessment of Function Annotation (CAFA) , have dePLOS Computational Biology. http://dx.doi.org/10.1371/journal.pcbi.1004926Novel computational protocols for functionally classifying and characterising serine beta-lactamases. David Lee, Sayoni Das, Natalie L. Dawson, Dragana Dobrijevic, John Ward, Christine Orengo. Published 22 Jun 2016. PLOS Computational Biology. http://dx.doi.org/10.1371/journal.pcbi.1005001Isofunctional protein subfamily detection using data integration and spectral clustering. Elisa Boari de Lima, Wagner Meira Júnior, Raquel Cardoso de De Melo-Minardi. Published 27 Jun 2016. PLOS Computational Biology. http://dx.doi.org/10.1371/journal.pcbi.1005053Defining the product chemical space of monoterpenoid synthases. Boxue Tian, C. Dale Poulter, Matthew P. Jacobson. Published 12 Aug 2016. PLOS Computational Biology. http://dx.doi.org/10.1371/journal.pcbi.1005284An atlas of peroxiredoxins created using an active site profile-based approach to functionally relevant clustering of proteins. Angela F. Harper, Janelle B. Leuthaeuser, Patricia C. Babbitt, John H. Morris, Thomas E. Ferrin, Leslie B. Poole, Jacquelyn S. Fetrow. Published 10 Feb 2017.


The micronucleus assay, developed to assess DNA damage induced by noxious agents, supplies information on whether the damage is due to clastogenic or aneugenic action. Although it is the test that can be used to assess agents' toxicity, it cannot provide information on the molecular events that result in the induction of micronuclei. To study the molecular events, the combination of both microscopic and analytical techniques is required. Flow-sorting induced micronuclei, based on their DNA content, in combination with chromosomal FISH and other molecular techniques, may provide information on these events.


During the last years, the number of refugees around the world increased to about 22.5 million. The mental health of refugees, especially of unaccompanied minors (70% between the ages of 16 and 18 years) who have been exposed to traumatic events , is generally impaired with symptoms of post-traumatic stress disorder, depression, and anxiety. Several studies revealed (1) a huge variation among the prevalence rates of these mental problems, and (2) that post-migration stressors  might be at least as detrimental to mental health as the traumatic events in pre- and peri-flight. As psychotherapy is a limited resource that should be reserved for severe cases and as language trainings are often publicly offered for refugees, we recommend focusing on intercultural competence, emotion regulation, and goal setting and goal striving in primary support programs: Intercultural competence fosters adaptation by giving knowledge about cultural differences in values and norms. Emotion regulation regarding empathy, positive reappraisal, and cultural differences in emotion expression fosters both adaptation and mental health. Finally, supporting unaccompanied refugee minors in their goal setting and goal striving is necessary, as they carry many unrealistic wishes and unattainable goals, which can be threatening to their mental health. Building on these three psychological processes, we provide recommendations for primary support programs for unaccompanied refugee minors that are aged 16 to 18 years. Due to wars and disasters, the number of forcibly displaced people in 2016 increased to 65.6 million worldwide. These included 22.5 million refugees, of which 2.8 million were asylum seekers. Half of the refugees were underage and about 75,000 asylum applications were lodged by unaccompanied minors . Hence, For many years, concerns focused on URM’s well-being with regard to physical diseases and injuries that are caused by disasters and wars . HoweverThis high maintenance of mental health problems in URM during the first years after arrival might reflect post-migration stressors rather than the lasting impact of experienced traumatic events pre- or during the flight. Indeed, URM reported an increase in stressful life events within the first two years after arrival in the host country , which wIndeed, URM reported an increased amount of both acculturation-specific and general hassles as compared to their peers from the host country and their peers with immigrant background but without flight experience ,23. AssoIn addition to daily hassles, the amount of acculturative stress URM experience is influenced by their acculturation strategies ,18. AccuIn addition, acculturation is a process that occurs over time. According to Berry , there aWith significantly rising global diversity, intercultural competence (IC) is considered a significant aspect of acculturation, biculturalism, or multiculturalism . It reflKim , as the Among other factors that affect the levels of IC, emotion regulation might be particularly important as especially empathy may enable URM to reach a level of unconscious competence behavior and adapt to intercultural differences ,37 whileEmotion regulation can be described as a process of the person’s capability to evaluate, manage, experience, express and improve emotional reactions in a way that helps proper functioning . It leadEmotion regulation strategies both facilitate the acculturation process and are generally helpful to deal with traumatic experiences . For insRegarding refugees, they not only experience extreme fear or shame because of traumatic experiences, but also often have difficulties with emotion regulation and anger management ,50. In aIn addition to emotion dysregulation, post-migration living difficulties and the experience of traumatic events in refugees have been associated with difficulties in engaging in goal-directed behavior . Upon thSeveral motivational strategies have been developed in order to help people setting attainable goals and striving for them. For instance, the strategy of mental contrasting (MC) connects people’s wishes with obstacles impeding their fulfillment . During Despite the high number of resilient URM and factors influencing mental health other than pre-flight traumatic experiences, most clinical interventions addressing mental health of URM focus on PTSD . HoweverIn contrast, governmental or organizational programs mainly support acculturation processes. However, most programs focus on structural economic, educational, or health aspects, as for instance providing refugees with housing, schooling, or vocational education, or integration in the labor market ,54,55,68However, these integration courses are generally only loosely based on psychological concepts that are known to influence acculturation  and mental health. In addition, they often consider refugees as helpless individuals, who are not able to take care of their lives independently. Yet, on the contrary, refugees are mostly individuals with a strong intention to survive and pull through, who have come to the host country with their own wishes and goals . In addiTherefore, primary support programs, which are designed for groups of URM, should include both elements that directly aim to reduce sub-threshold psychopathologies as well as foster acculturation by integrating the URMs own goals. The reviewed literature on mental health and acculturation in URM revealed several key constructs—intercultural competence, emotion regulation, goal setting, and goal striving—that root in different lines of psychological research. While intercultural competence is mainly discussed in cultural and organizational psychology, emotion regulation is a key construct in clinical psychology and developmental psychopathology, whereas goal setting and goal striving are primarily rooted in social and motivational psychology. Consequently, so far, no unifying framework exists that could guide the development of primary support programs for URM. As sensitivity to cultural differences is the basis for successful intercultural communication and IC in general , traininBased on Gross’ model for emotion regulation , improviHence, as emotion regulation capabilities are generally reduced in refugees and better emotion regulation strategies foster mental health and the acculturation process, primary support programs for URM should include elements of emotion regulation.Mental contrasting is a common strategy of goal setting and has been successfully applied to various wishes  and populations, including depressive patients  and chilHence, as URM often carry unrealistic expectancies and wishes towards their life in the host country, primary support programs for URM should help them in setting realistic goals, disengage from unattainable or unrealistic wishes, and strive efficiently towards goal attainment. The strategies of MC and II might be suited for support programs for URM, in particular, as they have proven their value for different populations and goals, including children and adolescents, as well as people suffering from internalizing and externalizing problems. In addition, the strategies are easy to administer and learn. Unaccompanied refugee minors are at a special risk for mental health problems due to traumatic experiences pre or during their flight as well as difficulties in the acculturation process. So far, support programs for URM either mainly focus on their traumatic experiences and resulting PTSD or on structural aspects as language and integration courses, integration into the schooling system or the labor market. However, these programs do not directly address psychological processes, which might offer URM resources, they might be able to use subsequently during their acculturation process. Primary support programs, which include elements of IC, emotion regulation, and goal setting and goal striving, therefore fill a gap. These elements are easy to integrate into the existing programs and can be applied in group settings, thereby reducing costs in terms of time or money. However, training material will need to be translated into the different languages of the URM currently coming into the host countries and validation studies should be carried out to ensure the material is culturally appropriate. Moreover, for larger groups it might be advisable to use translators and interpreters, who are more familiar with the refugees’ cultural backgrounds and have some experiences related to trainings or psychotherapies for refugees . Due to While about 70% of the URM are between 16 and 18 years , it is uIn addition, longitudinal studies are needed in order to evaluate the effectiveness of these program elements and whether the elements affect adaptation and mental health independently of each other or are interrelated. Furthermore, future research needs to analyze the personal characteristics of URM or the program conductors, which might act as potential moderators for the success of primary support programs. Finally, the three described elements influencing mental health and adaptation are not exclusive and other constructs should be evaluated with regard to trainability and their supporting effects.


To review the current status of positron emission tomography (PET) molecular imaging research of levodopa-induced dyskinesias (LIDs) in Parkinson’s disease (PD).Recent PET studies have provided robust evidence that LIDs in PD are associated with elevated and fluctuating striatal dopamine synaptic levels, which is a consequence of the imbalance between dopaminergic and serotonergic terminals, with the latter playing a key role in mishandling presynaptic dopamine release. Long-term exposure to levodopa is no longer believed to solely induce LIDs, as studies have highlighted that PD patients who go on to develop LIDs exhibit elevated putaminal dopamine release before the initiation of levodopa treatment, suggesting the involvement of other mechanisms, including altered neuronal firing and abnormal levels of phosphodiesterase 10A.Dopaminergic, serotonergic, glutamatergic, adenosinergic and opioid systems and phosphodiesterase 10A levels have been shown to be implicated in the development of LIDs in PD. However, no system may be considered sufficient on its own for the development of LIDs, and the mechanisms underlying LIDs in PD may have a multisystem origin. In line with this notion, future studies should use multimodal PET molecular imaging in the same individuals to shed further light on the different mechanisms underlying the development of LIDs in PD. More than 50 years since its discovery , 2, levoThe mechanisms underlying LIDs are still unclear. Levodopa induces sharp increases in striatal dopamine levels, which are particularly elevated in PD patients who experience LIDs . HoweverMolecular imaging modalities are able to identify minimal alterations at the nanomolecular level, and this is a prerequisite to understand subtle changes in brain activity , 18. PosThis review describes the current status of PET molecular imaging of LIDs, and its relation with the underlying mechanisms of PD.Multiple components of the network between basal nuclei and cortex have been recognized as a substrate for the development of LIDs. At the molecular level, changes in signal transduction and neurotransmission occurring in specific populations of neurons have been linked to the emergence of LIDs .l-amino acid decarboxylase (AADC), such as serotonergic and noradrenergic terminals, which do not possess the molecular machinery to properly control the release of dopamine. This results in higher swings in synaptic levels of dopamine and pulsatile stimulation of postsynaptic receptors located on striatal projection neurons raclopride, a D2 receptor antagonist radioligand which competes with endogenous dopamine for D2 receptor binding. Changes in D2 receptor availability, as reflected by a reduction of baseline [11C]raclopride levels after levodopa administration, allows indirect measurement of synaptic dopamine release. PD patients with LIDs showed increased dopamine release after the administration of levodopa compared with PD patients with stable response to levodopa [11C]raclopride levels has also been correlated with worse LIDs fluorodopa ([18F]FDOPA), [18F]dihydrotetrabenazine, and [18F]fluoropropylcarbomethoxyiodophenylnortropane ([18F]FP-CIT) or [11C]methylphenidate ([11C]MP) are reliable tools to assess in vivo striatal AADC activity, the density of vesicular monoamine transporter 2 (VMAT2), and the availability of presynaptic dopamine transporters (DATs). The AADC catalyzes the decarboxylation of levodopa to dopamine, the VMAT2 is the vesicular transporter responsible for the transport of dopamine from the cytoplasm into secretory vesicles and a marker of dopamine storage in the dopaminergic terminals, and the DAT is responsible for the high-affinity uptake of dopamine from the synaptic cleft, respectively [PET ligands for the presynaptic dopaminergic system such as [ectively .18F]FDOPA PET, is lower in PD patients, and correlates with the severity of motor symptom rigidity and bradykinesia [18F]FDOPA PET to measure the effective distribution volume ratio, as a marker of dopamine release, and who were followed up to the development of LIDs [18F]FDOPA uptake compared with PD patients with a stable response to levodopa [18F]FDOPA in the serotonergic and noradrenergic terminals, which possibly reflects a compensatory upregulation of AADC, has also been shown, and in the globus pallidus internal segment, which possibly reflects dopamine storage capacity [18F]dihydrotetrabenazine showed that in advanced PD patients, lower VMAT2 density in the caudate, putamen and substantia nigra was correlated with the severity of motor complications [18F]FDOPA uptake in the globus pallidus internal segment is lost with the progression of the disease in advanced PD patients with LIDs propyl-9-hydroxynaphthoxazine PET, compared with PD patients with stable response to levodopa FP-CIT PET at the time of diagnosis, showed that lower putaminal DAT levels at baseline was a predictor of the development of LIDs at the 3-year follow-up. A study with [11C]MP PET showed that advanced PD patients with motor fluctuations and LIDs had reduced DAT expression in the putamen compared with advanced PD patients with motor fluctuations but no LIDs SCH23390 PET and [11C]raclopride PET, respectively dimethyl-2-(2-amino-4-cyanophenylthio)benzylamine ([11C]DASB) are reliable tools to assess in vivo serotonergic terminals [11C]raclopride challenge, compared with PD patients with stable responses to levodopa. Oral administration of the serotonin receptor type 1A agonist buspirone, a presynaptic modulator of the serotonergic system, before levodopa administration reduced levodopa-evoked striatal synaptic dopamine release and attenuated LIDs [18F]FP-CIT PET, compared with non-dyskinetic PD patients; however, the [11C]DASB to [18F]FP-CIT binding ratio  was higher in the putamen of PD patients with LIDs [11C]DASB PET. PD patients with LIDs had relative preservation of serotonergic terminals in the globus pallidus and increased pallidal dopamine release, measured with levodopa challenge and PET methoxyphenoxy)benzyl)morpholine or [11C]methyl reboxetine are not reliable tools to assess in vivo noradrenaline transporter activity methyl-4-piperidyl acetate ([11C]MP4A) and N-[11C]methylpiperidinyl propionate ([11C]PMP) are reliable tools to assess in vivo acetylcholinesterase activity [11C]MP4A and [11C]PMP have demonstrated 11–12% decreases in cortical and subcortical acetylcholinesterase activity in PD patients without dementia A-85380 and N-[11C]methylpiperidyl benzilate ([11C]NMPB) are reliable tools to assess in vivo nicotinic and muscarinic receptors respectively. A PET study using [18F]A-85380 was performed in PD patients and showed reduced levels of nicotinic receptors in the striatum and substantia nigra compared with the levels in controls [18F]A-85380 levels and disease severity, but LIDs have not been evaluated in detail. No PET studies have been performed in humans investigating the role of the cholinergic system in LIDs with [18F]A-85380, [11C]NMPB or other tracers; thus, this component needs further investigation.The main postsynaptic component of cholinergic pathways is from cholinergic neurons intrinsic to the striatum . Global patients , 77. Thicontrols . There wN-methyl-d-aspartate (NMDA) receptors, which include the NR1, NR2A and NR2B subtypes [11C]CNS51619 are reliable tools to investigate in vivo the glutamatergic system [11C]CNS51619 uptake in the caudate, putamen and precentral gyrus compared with PD patients without LIDs, suggesting that dyskinetic patients may have abnormal glutamatergic transmission in motor areas following levodopa administration 8-dicyclopropylmethyl-1-methyl-3-propylxanthine and [7-methyl-11C]-(E)-8--1,3,7-trimethylxanthine or [11C]SCH442,416 are reliable tools to measure in vivo A1A and A2A receptors, respectively. Two PET studies investigating the adenosinergic system have been performed in PD patients with LIDs diprenorphine, a non-selective opioid receptor PET ligand, is a reliable tool to assess in vivo the opioid system [11C]diprenorphine in PD patients, and showed reduced striatal, thalamic and cingulate opioid receptor binding and increased prefrontal opioid receptor binding in PD patients with LIDs compared with PD patients with stable response to levodopa [11C]diprenorphine levels and PD severity, disease duration, or duration of levodopa treatment [Three opioid receptors subtypes  are involved in regulating dopamine functions . [11C]did system . The mailevodopa . No corrreatment . Furtheri/o proteins in the direct pathway, and they counteract D2 inhibition of adenylate cyclase coupling with Gs proteins in the indirect pathway, thus having a key role in the control of movement [18F]MK-9470 are a reliable tool to measure in vivo CB1 receptors [18F]MK-9470, which demonstrated a significant decrease in CB1 receptor availability in the substantia nigra compared with healthy controls. No correlation was found between reductions in CB1 receptor levels and the severity of LIDs methoxyphenyl)-3-((R)-1-phenylethylamino)-1-(4-trifluoromethylphenyl)pyrrolidin-2-one IMA107 has been used to quantify PDE10A expression in vivo in moderate–advanced PD patients [PDE10A is mainly expressed in striatal medium spiny neurons, where it regulates the cAMP/PKA/DARPP-32 signalling cascade, thus plays a key role in the regulation of the global striatal output and in promoting neuronal survival , 95. PDEpatients . PD patipatients . This prStudies have provided robust evidence that LIDs in PD are associated with elevated and fluctuating striatal dopamine synaptic levels arising from an imbalanced dopaminergic to serotonergic terminal ratio, with the latter playing a key role in mishandling presynaptic dopamine release. However, other non-dopaminergic systems and pathways such as the glutamatergic, adenosinergic and opioid systems and phosphodiesterase 10A may play important roles in the development of LIDs in patients with PD. The implementation of novel PET ligands is warranted to unveil unexplored mechanisms of underlying pathophysiology of LIDs, and applications of multimodal PET molecular imaging approaches combining different tracers may shed further light on the mechanisms underlying the development of LIDs in PD.


Human herpes virus-8, a γ2-herpes virus, is the aetiological agent of Kaposi sarcoma. Recently, Kaposi's sarcoma cases have increased in Zambia. However, the diagnosis of this disease is based on morphological appearance of affected tissues using histological techniques, and the association with its causative agent, Human Herpes virus 8 is not sought. This means poor prognosis for affected patients since the causative agent is not targeted during diagnosis and KS lesions may be mistaken for other reactive and neoplastic vascular proliferations when only histological techniques are used. Therefore, this study was aimed at providing evidence of Human Herpes virus 8 infection in Kaposi's sarcoma tissues at the University Teaching Hospital in Lusaka, Zambia.One hundred and twenty suspected Kaposi's sarcoma archival formalin-fixed paraffin-wax embedded tissues stored from January 2013 to December 2014 in the Histopathology Laboratory at the University Teaching Hospital, Lusaka, Zambia were analysed using histology and Polymerase Chain Reaction targeting the ORF26 gene of Human Herpes virus 8.The predominant histological type of Kaposi's sarcoma detected was the Nodular type (60.7%) followed by the plaque type (22.6%) and patch type (16.7%). The nodular lesion was identified mostly in males  than females  (p=0.041). Human Herpes virus 8 DNA was detected in 53.6% (45/84) and mostly in the nodular KS lesions  (p=0.035).The findings in this study show that the Human Herpes virus-8 is detectable in Kaposi's sarcoma tissues, and, as previously reported in other settings, is closely associated with Kaposi's sarcoma. The study has provided important baseline data for use in the diagnosis of this disease and the identification of the virus in the tissues will aid in targeted therapy. Human herpes virus-8 (HHV-8), a γ2-herpes virus, also known as Kaposi sarcoma-associated herpesvirus (KSHV), is the aetiological agent of Kaposi sarcoma(KS) –3 and isKS is now the most frequently reported malignant skin tumour in some areas of Africa and was endemic in Africa even before the advent of Human Immunodeficiency Virus (HIV) infection , 16. It Specimens: The study was a laboratory-based retrospective study on 120 archival formalin-fixed paraffin-embedded (FFPE) KS tissues. It study was conducted at the University Teaching Hospital (UTH) in the Department of Pathology and Microbiology, Histopathology Laboratory in Lusaka. The UTH is a tertiary referral and teaching hospital with a bed capacity of about 1,664. The hospital has about 11 departments namely: Obstetrics and Gynaecology, Paediatrics, Surgery, Community medicine, Pathology, Radiology, Physiotherapy, Pharmacy and Blood bank. It is the biggest referral hospital and the centre for all histopathology diagnostic work in Zambia.Histological examination: Sections of tissue were cut at 6μm on a Shandon Finesse 325 microtome . The tissue sections were then stained using Haematoxylin and Eosin staining and examined by a qualified pathologist. The KS lesions were grouped into patch, plaque and nodular histological stages.DNA extraction: Tissue sections were cut as described above. Up to 30mg of tissue sections was placed in a sterile 1.5ml microfuge tube. DNA was extracted using the EZNA Tissue DNA Extraction Kit  for paraffin-embedded tissue according to the manufacturer´s protocol. The DNA was eluted in 50μl volumes, and then stored at -20°C until required.PCR amplification and detection of HHV-8 ORF26 gene: DNA extracted was used for the detection of the HHV-8 sequences. Nested PCR was performed using two sets of primers, KS-1(5'-AGCCGAAAGATTCCACCAT-3') and KS-2 (5'-TCCGTGTTGTCTACGTCCAG-5'), KS-4(5'-CGAATCCAACGGATTTGACCTC-3') and KS-5(5'-CCCATAAATGACACATTGGTGGTA-3') amplifying the ORF26 gene of the HHV-8 genome. The 2 sets of primers were used for screening the DNA for HHV-8. The first round PCR reactions were performed in a final volume of 25μl of which 2μl was genomic DNA, 1X PCR buffer, 3.5 mM MgCl2, 0.2mM of deoxynucleoside triphosphates, and 0.2 U of Invitrogen Taq DNA polymerase , 2.5μl of each primer on a Gene amplification 2700 Thermocycler . Primers were combined at a final concentration of 0.2μM. For the second-round PCR, 2μl of the first-round PCR products was used as the template DNA. The cycling procedure for the KS-1, KS-2 primer pair was 94°C for 5 minutes, then 35 cycles of 94°C for 30 seconds, 58°C for 30 seconds, 72°C for 30 seconds and 1 final extension cycle of 7 minutes at 72°C. For the KS-4 and KS-5 primers, the amplification conditions were similar except the annealing temperature was at 53°C. The PCR products were analysed by electrophoresis on a 1.5% agarose gel containing ethidium bromide (10μg/ml in distilled water) and were visualized under ultraviolet light alongside a 50bp DNA ladder using the Biotop SC-645 Gel Documentation system . A known KS case was used as a positive control and a negative control containing nuclease-free without DNA was always included. The expected product size was 233bp.As an internal control, the extracted DNA was subjected to PCR using primers for human β-actin (Forward 5'-GCC ATG TAC GTT GCT ATC C-3' and Reverse 5'-CCG CGC TCG GTG AGG-3') with the following conditions: initial denaturation of 94°C for 1 minute, 35 cycles of 95°C for 30 seconds, 62°C for 1minute, 72°C for 1 minute and 1 final extension cycle of 10minutes at 72°C. The PCR products were also analysed by electrophoresis on a 1.5% agarose gel containing ethidium bromide (10μg/ml) and visualised under ultraviolet light alongside the 50bp DNA ladder. The expected product size was 200bp.Statistical analysis: Statistical analysis was performed using GraphPad Prism Software Version 6 . Logistic regression was used to determine associations among HHV-8, histological types of KS, gender and age. Chi-square and Fisher's exact test were used to determine associations among histological types of KS, gender and age. A p-value of less than 0.05 was chosen to indicate statistical significance.Ethics considerations: This study was laboratory-based with no direct contact with patients. Permission to use the archived histopathology tissues was sought from the Head of the Department of Pathology and Microbiology at the University Teaching Hospital. Ethics approval for the research was sought from the University of Zambia Biomedical and Research Ethics Committee (UNZABREC) (Ethics clearance number: 002-04-14/B).Histology: Out of the 120 suspected FFPE KS tissues analysed, 70% (84/120) were confirmed KS cases in mostly males    A than f, 56/84)  B. The K, 56/84)  A. The n, 56/84)  B. The a, 56/84) .Detection of HHV-8 ORF26 gene: The presence of HHV-8 DNA was confirmed by running nested PCR products on a 1.5% agarose gel and a product size of 233bp was observed as shown in HHV-8 has been proven to be the causal agent of all forms of KS , 21. TheThis study also showed that the occurrence of KS was proportionally higher in men than in women. Men presented with more nodular lesions of KS compared to women. These findings are similar to studies carried out in other sub-Saharan countries , 26. A UIn this study, HHV-8 detection was higher in the nodular lesions of KS than other lesions. The nodular lesions were more likely to harbour HHV-8 DNA (OR=1.43 95% CI 0.06-2.04 p=0.035) than the other histological types of KS. These findings were consistent to those in a US study in which higher levels of HHV-8 DNA were also reported in the nodular stage compared to the patch and plaque stages . Data obData generated in this study shows that KS lesions consist of three histological types: patch, plaque and nodular lesions. Of the three, the nodular lesion was the most common (60.7%) and was mainly identified in males than in females, supporting the notion that KS is a male- associated disease. Another significant finding was the detection of HHV-8 DNA in more than half of the KS tissues analysed, and this was mostly detected in the nodular lesions. Our data corroborate those from other studies suggest a role of HHV-8 in KS pathogenesis. We propose detection of HHV-8 in tissues by PCR as a way of making a definitive diagnosis for KS, especially in early vascular proliferations when the characteristic histopathological features are not present.It is already known that HHV-8 is aetiologically linked to KS;HHV-8 infection is more common in males than females.This study is the first of its kind to be done in Zambia and hence adds to the knowledge that HHV-8 is detectable in FFPE tissues of KS;The nodular KS are the most common type of KS in Zambia.The authors declare no competing interest.


The purpose of this study was to review the recent randomised controlled trials of coronary computed tomography angiography (CCTA) for patients with stable coronary artery disease.The initial results and subsequent papers from the SCOT-HEART (Scottish COmputed Tomography of the HEART) and PROMISE  trials have shown that CCTA is a safe and appropriate addition to standard care or alternative to functional testing. The SCOT-HEART study showed that CCTA changes diagnoses, improves diagnostic certainty, changes management, leads to more appropriate use of invasive coronary angiography, and reduces fatal and non-fatal myocardial infarction. A meta-analysis of the four randomised controlled trials showed that CCTA leads to a major reduction in myocardial infarction in patients with stable chest pain.CCTA is now an established technique for the assessment of coronary artery disease. Recent ‘test and treat’ randomised controlled trials have shown that CCTA guided changes in management can improve clinical outcomes. Coronary computed tomography angiography (CCTA) is now an established technique for the assessment of patients with suspected coronary artery disease (CAD). It has an excellent diagnostic accuracy for the identification of CAD, with a high negative predictive value . Large-sA normal CCTA is associated with a good prognosis in large registry studies of patients with suspected CAD –4. Both The PROMISE  trial recruited 10,003 symptomatic stable outpatients who were due to undergo non-invasive investigation for suspected CAD ••. PartiThe mean age of the participants was 61 years, 53% were female and the pre-test probability of obstructive CAD was 53%. Over one quarter of patients 27%) presented with a primary symptom other than chest pain, such as breathlessness or fatigue. The characterisation of chest pain was typical angina for 12%, atypical angina for 78% and non-anginal chest pain for 11% (Table 7% presenP = 0.049). However, at 25 months of follow-up, there was no difference in the primary outcome between the two groups . Thus, PROMISE showed that CCTA is a safe alternative to functional testing in a low-risk population with similar outcomes in both groups after 2 years of follow-up.The primary outcome of the PROMISE study was a composite of all-cause mortality, myocardial infarction, hospitalisation for unstable angina and major complications of cardiovascular procedures or diagnostic testing. At 12 months of follow-up, the risk of death or non-fatal myocardial infarction was lower in the CCTA group than in the functional imaging group (hazard ratio (HR) 0.66, 95% confidence interval (CI) 0.44–1.00, The SCOT-HEART (Scottish COmputed Tomography of the HEART) trial randomised 4146 outpatients with suspected angina due to CAD to standard care or standard care plus CCTA . ParticiThe mean age of participants was 57 years, 44% were female and the pre-test probability of obstructive coronary heart disease was 47%. The presence of typical angina was higher in the SCOT-HEART trial as compared to the PROMISE trial (35 vs 12% of participants). Atypical angina was the presenting complaint in 24% of patients (cf. 78% in PROMISE) and 41% (cf. 11%) had non-anginal chest pain (Table p < 0.0001). CCTA improved the certainty of the diagnosis for both the presence of CAD and the diagnosis of angina due to (CAD) . CCTA increased the frequency of the diagnosis of CAD (relative risk (RR) 1.09, 95% CI 1.02–1.17, P = 0.0172), but tended to decrease the frequency of the diagnosis of angina due to significant CAD .The primary endpoint of the SCOT-HEART trial was the certainty of the diagnosis angina pectoris secondary to significant CAD at 6 weeks Table . At 6 weP = 0.0527) [P = 0.020) [At 1.7 years of follow-up, the CCTA group had a 38% lower rate of fatal and non-fatal myocardial infarction as compared to the control group, but this difference did not quite reach statistical significance  ••. The o= 0.020) ••.There are two other smaller randomised controlled trials which have recently assessed CCTA in patients with stable chest pain: the CAPP trial • and a sP = 0.34)  Table . The priP = 0.04), and 44% were female . • (Table le Table . Pre-tesP = 0.038), but there was no difference in all-cause mortality [A meta-analysis has combined the primary results of PROMISE, SCOT-HEART, CAPP and the study by Min et al. •. It ideortality •. This 3P < 0.0001) [P = 0.451). In the study by Min et al., there were also similar rates of ICA between the CCTA and the MPS groups [In the SCOT-HEART trial, CCTA led to a change in the planned investigations in 15% of participants, compared to 1% in the standard care group ( 0.0001) ••. For sS groups •. HoweveS groups ••, 11••.P < 0.0001). This included an increase in the use of preventative therapies  and decrease in the use of anti-anginal therapies [In the SCOT-HEART study, treatment was changed in 23% of patients in the CCTA group compared to 5% in the standard care group  [P = 0.0611) [In the PROMISE trial, there was an increase in the numbers of patients going on to have revascularisation after invasive coronary angiography in the CCTA group  ••. In th 0.0611) ••. ThereIn both the SCOT-HEART and the PROMISE trials, there was a small increase in cost in the CCTA group. In the PROMISE study, the mean cost difference at 90 days was $254  , and in In SCOT-HEART, PROMISE and CAPP trials, symptoms improved in both groups of patients indicating general satisfaction after attendance at the RACPC or cardiology outpatient clinic , 17. In P < 0.001). However, as 33% of patients in the functional testing group did not have any radiation exposure at all, and therefore, the median cumulative radiation exposure per patient was lower in the CCTA group (10.0 vs 11 mSv). In the SCOT-HEART trial, the median radiation dose for CCTA and non-contrast coronary artery calcium score was 4.1 (interquartile range 3.0–5.6) mSv. More than one third of the radiation exposure in the SCOT-HEART trial was attributed to the coronary artery calcium score. The additional benefit of performing a non-contrast scan for calcium scoring in patients who are undergoing CCTA is low, and indeed, in many cases, this additional examination is not required. The mean radiation dose in the CCTA group of the CAPP trial was 5.37 mSv. In the study by Min et al., the CCTA group had a significantly lower radiation dose than the MPS group . CCTA can now be performed at a low radiation dose for patients with a range of heart rates and body size  vs 10.1 mSv, P = 0.0001) [In addition to assessing stenosis severity, CCTA can assess the constituents and morphology of atherosclerotic plaques. Motoyama et al. identified that the presence of low attenuation plaque, positive remodelling and spotty calcification were associated with an increased risk of acute coronary syndrome . A meta- 0.0001) . HoweverCardiac CT can assess myocardial perfusion using static or dynamic techniques with good diagnostic accuracy compared to other imaging modalities . ComputaCCTA is a safe diagnostic test that can be performed at a low radiation dose and is now widely available. Recent clinical trials have established the role of CCTA in the assessment of patients with suspected CAD. The use of CCTA not only changes the diagnosis but it also alters management that ultimately reduces the future risk of fatal and non-fatal myocardial infarction. National guidelines, such as the UK NICE guidelines, have recently changed to incorporate CCTA as the first-line test in stable chest pain  and other international guidelines are likely to follow suit in time.