One study suggested a correction formula for femoral venous access that markedly reduced the bias for GEDVI. Therefore, the last PiCCO-algorithm requires information about the CVC-position which suggests a correction of GEDV for femoral access. However, a recent study demonstrated inconsistencies of the last PiCCO algorithm using incorrected GEDV to calculate CFI despite obvious correction of GEDV. Nevertheless, this study was based on mathematical analyses of data displayed in a total of 15 patients equipped with only a femoral, but not with a jugular CVC. Therefore, this study compared CFI derived from the femoral indicator injection TPTD to data derived from jugular indicator injection in 28 patients with both a jugular and a femoral CVC.Cardiac function index (CFI) is a trans-pulmonary thermodilution (TPTD)-derived estimate of systolic function. CFI is defined as the ratio of cardiac output divided by global end-diastolic volume GEDV (CFI = CO/GEDV). Several studies demonstrated that the use of jugular venous gold standard access and the femoral access with and without information about the femoral indicator injection to evaluate, if correction for femoral GEDV also pertains to CFI. ).28 ICU-patients with PiCCO-monitoring were included. Each dataset consisted of three triplicate TPTDs using the -1; p<0.001). Similarly, CFI_fem_cor was significantly lower than CFI_jug . This is explained by the finding that CFI_fem_uncor was not different to CFI_fem_cor . This suggests that correction for femoral CVC does not pertain to CFI. Calculative correction of CFI_fem_uncor by multiplying CFI_fem_uncor by the ratio GEDVI_fem_uncor/GEDVI_jug resulted in CFI_fem_uncor_form which was slightly, but significantly different from the gold standard CFI_jug . The agreement of measurements classified in the same category of CFI (decreased (<4.5), normal (4.5–6.5) and increased (>6.5 min-1)) was high for CFI_jug and CFI_fem_uncor_form . By contrast, the agreement with CFI_jug was significantly lower for CFI_fem_cor  and CFI_fem_uncor .CFI_fem_uncor was significantly lower than CFI_jug (4.28±1.70 vs. 5.21±1.91 minWhile the last PiCCO algorithm obviously corrects GEDVI for femoral indicator injection, this correction is not applied to CFI. Therefore, femoral TPTD indicator injection results in substantially lower values for CFI compared to TPTD using a jugular CVC. Necessarily, uncorrected CFI-values derived from femoral TPTD are misleading and have to be corrected. Accurate haemodynamic monitoring is essential for the diagnosis and therapeutic management of critically ill patients with circulatory failure . DiffereBased on their mathematical derivation, CFI (CO/GEDV) and GEF (4*SV/GEDV) necessarily depend on an accurate determination of GEDV. However, several studies demonstrated that GEDV is markedly overestimated in case of using a femoral venous access for TPTD-indicator injection instead of a jugular or subclavian access –16. ConsAnother recent study in patients equipped with both jugular and femoral CVCs demonstrated that the last PiCCO-algorithm corrects GEF, but not PVPI which resulted in a substantial underestimation of PVPI in case of femoral indicator injection . HoweverTherefore, we have compared in the present study CFI values of 28 patients equipped with both jugular and femoral CVCs: Two triplicate measurements with femoral indicator injection with and without giving the information of femoral indicator injection were compared to the gold-standard of CFI derived from jugular indicator injection.This prospective observational study was conducted in a ten-bed general ICU at a university hospital between October 6, 2016 and March 31, 2017. The study was approved by institutional review board approved . Written informed consent was obtained by all patients (18 years of age or older) or their legal representatives. All patients had to be equipped with PiCCO device and with both jugular and femoral catheter. The indication for PiCCO monitoring was made independently from the study by the ICU physician in charge None of the patients had been included in one of the previous studies or databases comparing TPTD-parameters derived from jugular to femoral indicator injection , 17–19. The abbreviations and nomenclature of haemodynamic parameters used in this paper are summarized in with (TPTD_fem_cor) or without (TPTD_fem_uncor) the information about the femoral indicator injection. This was done to evaluate, if correction for femoral GEDV pertains to CFI_fem.28 datasets including triplicate TPTD with 15 ml cold saline solution were recorded in 28 patients equipped with both jugular and femoral CVC. The jugular venous access was used as the gold standard TPTD_jug. Furthermore, two triplicate TPTDs were performed via the femoral access The three TPTDs were performed in a random order with the intention to avoid a systematic bias by repeated triplicate TPTDs with a total volume of 9*15 ml.All measurements were performed in patients equipped either with conventional jugular and femoral CVC or conventional CVC and a dialysis catheter irrespective of the study. According to the local standard CVCs or dialysis catheters were inserted in different positions (one in the superior vena cava and the other one in the inferior vena cava).st indicator injection, since the larger volume of the dialysis catheters (up to 1.6 ml) might result in a loss of indicator  and in a consecutive overestimation of volumetric parameters for the 1st of TPTD-measurement. Femoral venous catheters were completely  inserted under ultrasound guidance. The position of the tip was controlled (and corrected) according to X-ray for all jugular, but not for the femoral venous catheters.A 5-lumen CVC  with a maximum intravascular length of 20 cm and a diameter of 3.15 mm (9.5 French (Fr)) or a Gambro Gam Cath Dolphin dialysis-catheter  was used for TPTD indicator injections. For femoral access dialysis catheters with a length of 250 mm and a diameter of 13 Fr were used. For jugular RRT access we used catheters with a length of 150–175 mm and a diameter of 13 F. The dialysis catheters were prefilled with ice cold saline immediately before the 1TPTD was performed as previously described , 20, 21 All data were controlled for input data error. Continuous variables are expressed as mean±standard deviation. Categorical variables are expressed as percentages. Wilcoxon-test for paired samples was used to compare continuous variables.Bland-Altman analysis was used for the analysis of the agreement between CFI derived from jugular vs. femoral venous catheter sites for CFI and to compute the percentage error (PE).-1), normal (4.5–6.5 min-1) and increased (>6.5 min-1)) derived from different measurements was primarily analyzed using Fisher´s exact test (“agreement yes or no”). Additionally, we calculated kappa-statistics and Kendall´s coefficient of correlation.The agreement of classification of CFI .-1 and significantly lower values of 3.8±1.6 min-1 derived from uncorrected femoral indicator injection, using an online statistical power calculator  Sample sizes of n = 12 and n = 16 would provide statistical powers of 80% and 90% respectively [-1 instead of 1.6 min-1 in the study by Beitz et al.. This resulted in a sample size of n = 28 to provide a statistical power of 90% and an alpha-error of 5% (two-tailed test).The sample size calculation based on the finding of the previous study by Beitz et al. with CFI-values calculated for jugular indicator-injection of 5.1±1.8 minectively . Howeverectively . A large-1; p<0.001; CFI_fem_uncor was significantly lower than CFI_jug  was in the same range as the ratio of GEDVI_fem_uncor/GEDVI_fem_cor .The relation of CFI_jug/CFI_fem_uncor  [Therefore, CFI_fem_form was calculated by correcting CFI_fem_uncor by multiplication of CFI_fem_uncor with the ratio GEDVI_fem_uncor/GEDVI_fem_cor using the recently suggested correction formula for GEDVI_fem based on uncorrected GEDVI and cardiac index (CI) derived from femoral indicator injection (GEDV_fem_uncor and CI_fem_uncor) as well as on ideal bodyweight  :GEDVI_fConsequently, for ex-post-correction of CFI_fem_uncor we calculated CFI_fem_uncor_form by multiplying CFI_fem_uncor with the ratio GEDVI_fem_uncor/GEDVI_fem_cor:-1; p = 0.024; -1 and a percentage error of 29.6%. The CV-values were comparable for CFI_fem_uncor_form and CFI_jug .CFI_fem_uncor_form was slightly, but significantly different from the gold standard CFI_jug .Despite a slightly significant difference between CFI_fem_uncor_form and CFI_jug with questionable clinical relevance, ex-post correction of CFI_uncor resulted in significantly lower amount of the bias |CFI_fem_uncor_form–CFI_jug| vs. |CFI_fem_uncor–CFI_jug|  for the “gold-standard” CFI_jug vs. the classifications according to CFI_fem_cor, CFI_fem_uncor and CFI_fem_uncor_form, respectively , which was not significant different to CFI_jug , but significantly higher compared to CFI_fem_uncor (p = 0.0019) and CFI_fem_cor (p<0.001).Furthermore, kappa-statistics and Kendall´s coefficient tau-b confirm a markedly better agreement of CFI_fem_uncor_form with CFI_jug  compared to CFI_fem_uncor  and CFI_fem_cor .TPTD-derived CFI is a bedside surrogate of LV systolic function. CFI is strongly associated with echocardiography-derived LVEF and facilitates guidance of inotropic therapy and fluid management. Repeated CFI measurement is readily available and independent of the examiner. Changes in CFI over time provide dynamic information that might be superior to single measurements, particularly when interpreted in the light of a clinical situation. According studies with their findings are summarised in femoral CVC for indicator injection, compared to the gold standard of jugular or subclavian injection. Interestingly, a similar phenomenon was found in case of misplacement of subclavian central venous catheter tip into the jugular vein [However, the validity of CFI calculation relies on the accurate determination of CO and GEDV. Several recent studies suggest marked overestimation of GEDV/GEDVI and CFI in case of using a lar vein .One study suggested a correction formula for GEDVI derived from femoral indicator injection. This correction is based on GEDVI and CI obtained from femoral access and on ideal bodyweight . SeveralThis study demonstrates that the correction formula for femoral venous access is not applied to correct CFI. The resulting underestimation of this value would have had a consequence for around 50% of our patients as demonstrated by the wrong classification of CFI in 14 out of 28 measurements. Therefore, measurement of CFI in patients with femoral venous access for indicator injection at present is misleading and has to be replaced by echocardiography as long as the correction formula is not implemented in the TPTD-algorithm. This problem might also apply to the other commercially available TPTD-device EV-1000 , since at least one study suggests that this device does not correct GEDVI, PVPI and GEF for femoral indicator injection [femoral central venous access echocardiography should be performed to assess left ventricular contractility. Irrespective of the central venous access, echocardiography enables to exclude isolated right heart failure which might impede the use of CFI and GEF also in case of a jugular or subclavian central venous access [Unless an appropriate correction is implemented in the PiCCO and the EV-1000, in patients with However, repeated echocardiography is time consuming and requires the continuous availability of experienced investigators. On the other hand, TPTD is straightforward and reliable even when performed by different investigators. Furthermore, it provides additional extra-cardiac parameters such as EVLWI and calibrates continuous measurement of CI, GEF and CFI.femoral venous access CFI has to be classified as misleading and may result in wrong therapeutic interventions due to substantial underestimation and wrong categorization of CFI.Although a correction formula for femoral venous access markedly reducing the bias for GEDVI has been published 7 years ago, and despite several studies gave hints for inconsistencies of the correction of GEDVI, PVPI, GEG and CFI, our data demonstrate that the most recent algorithm of the PICCO still does not apply this correction to CFI. Therefore, in patients with From a practical viewpoint, there are two options to overcome this dilemma in addition to the use of echocardiography:As demonstrated by this study, ex post correction by the previously suggested formula to correct GEDVI appropriately corrects CFI_fem with acceptable bias, percentage error and categorization according to clinical thresholds.Since this mathematical correction maybe cumbersome in clinical routine, GEF can be used instead of CFI. At least two studies suggest that GEF is appropriately corrected for femoral indicator injection by the most recent PiCCO algorithm , 19.This study included a small number of patients and has been conducted as a single-centre study. Furthermore, all measurements were performed in critically ill patients and not in healthy persons.femoral TPTD indicator injection results in substantially lower values for CFI compared to TPTD using a jugular CVC. Necessarily, uncorrected CFI-values derived from femoral TPTD are misleading and have to be corrected.While the last PiCCO algorithm obviously corrects GEDVI for femoral indicator injection, this correction is not applied to CFI. Therefore,


Surface modification of the Zr‐BTB structure was realized through the attachment of porphyrin‐based carboxylate ligands on the coordinatively unsaturated Zr metal sites in the MOF through strong Zr‐carboxylate bond formation. The functionalized MOF nanosheet, namely PCN‐134‐2D, acts as an efficient photocatalyst for 1O2 generation and artemisinin production. Compared to the 3D analogue (PCN‐134‐3D), PCN‐134‐2D allows for fast reaction kinetics due to the enhanced accessibility of the catalytic sites within the structure and facile substrate diffusion. Additionally, PCN‐134(Ni)‐2D exhibits an exceptional yield of artemisinin, surpassing all reported homo‐ or heterogeneous photocatalysts for the artemisinin production.2D metal–organic frameworks (2D‐MOFs) have recently emerged as promising materials for gas separations, sensing, conduction, and catalysis. However, the stability of these 2D‐MOF catalysts and the tunability over catalytic environments are limited. Herein, it is demonstrated that 2D‐MOFs can act as stable and highly accessible catalyst supports by introducing more firmly anchored photosensitizers as bridging ligands. An ultrathin MOF nanosheet‐based material, Zr‐BTB benzene), is initially constructed by connecting Zr MOFs are extended network structures constructed from inorganic metal nodes and organic ligands, typically resulting in 3D materials with inherent porosity.6 clusters and the tritopic carboxylate ligand 1,3,5‐tris(4‐carboxyphenyl)benzene (BTB), was selected as a catalyst support for our study.6 clusters that are hexacoordinate in the equatorial plane, thus leaving six pairs of terminal −OH−/H2O above and below the layer for functionalization of the material. The terminal −OH−/H2O in the structure can be replaced by carboxylate ligands, providing a facile approach for surface modification. Therefore, Zr‐BTB can be perceived as a MOF analogue of inorganic ZrP with enhanced tunability. In this work, 2D Zr‐BTB nanosheets with controlled thicknesses were synthesized by adjusting the interlayer van der Waals interactions. Porphyrin‐based ligands were subsequently anchored on the surface of the Zr‐BTB nanosheets by replacing the terminal −OH−/H2O on the Zr6 clusters. The resulting 2D material, PCN‐134‐2D, was utilized as a photocatalyst for the oxidation of dihydroartemisinic acid to produce artemisinin. When compared to the 3D counterpart (PCN‐134‐3D), the 2D nanosheets (PCN‐134‐2D) demonstrated improved reaction kinetics. This improvement was due to the more readily accessible catalytic centers that the high dispersibility of the individual nanosheets provided over that of the 3D PCN‐134 structure.With these strategies in mind, a previously reported 2D MOF based on Zr22.16 clusters with the tritopic Zr‐BTB linker into a ‐connected network. This arrangement allows for the formation of a coordinately unsaturated Zr6 cluster in the MOF. The coordinatively unsaturated Zr6 clusters endow unique tunability to the Zr‐BTB structure, with the terminal −OH−/H2O ligands being replaced by carboxylate ligands. This unique tunability allows for the functionalization of the Zr‐BTB nanosheets.FigureAmong the limited number of 2D Zr‐MOFs, Zr‐BTB stands out as a suitable catalyst support. This MOF was initially reported by Sun and co‐workers as interpenetrated nets, and was further studied by the Matzger and Zhao groups as a 2D material.This problem can be overcome by reducing the 3D MOF to a 2D regime. To obtain PCN‐134‐2D, a stepwise synthetic method was designed. The traditional synthesis of PCN‐134‐3D was carried out in a one‐pot reaction without the isolation of the Zr‐BTB intermediate , Zr‐BTB (10 mg), DMF (3 mL), terminal carboxylic acid ligand, and H2O were heated at 120 °C for 48 h to obtain the Zr‐BTB product. The resulting product was analyzed by powder X‐ray diffraction (PXRD) and scanning electron microscopy (SEM) to determine the structure and layer thickness. The addition of water significantly hindered the packing of the Zr‐BTB layers as indicated by the gradually reduction in diffraction intensity along the c‐direction in the PXRD pattern. When benzoic acid (BA) was used as the modulating agent, the peaks corresponding to the 001 and 111 crystal faces disappeared upon the addition of water to the reaction  and (111) diffraction peaks in the PXRD , whereas these peaks were not observed when bulkier modulating carboxylic acids were used . During the MOF synthesis, the modulating carboxylate acid linkers competitively coordinate with the metal cations, slowing down the crystal growth and allowing for the formation of highly crystalline products. The interaction between the metal cations and modulating carboxylates become diminished when bulky substituents are used due to steric hindrances, resulting in a weaker modulating effect. Therefore, bulky carboxylic acids such as PA and CA produced low crystallinity Zr‐BTB products . This is in line with the BET surface areas estimated by N2.31H‐NMR spectra of the digested samples  for the Zr‐BTB samples modulated by AA (12%), PA (11%), and CA (20%) were comparable to those of the samples modulated by FA (31%) and BA (32%). This was determined through thermogravimetric analysis (TGA) and indicated a greater degree of missing BTB defects in the structures . Based on the layer thickness, crystallinity, surface areas, and TCPP uptakes, the Zr‐BTB nanosheets synthesized using BA as a modulator followed by 0.5 mL water were selected for further studies. PCN‐134‐2D was obtained by the TCPP modification of aforementioned Zr‐BTB nanosheets.Porphyrin catalysts were attached to the Zr‐BTB nanosheets by incubating the as‐synthesized Zr‐BTB samples in solutions of DMF and TCPP at 100 °C. The TCPP uptakes into the structure, represented by TCPP/BTB ratios, were determined by the s Figure c. TCPP a2.4FigureThe structure and morphology of PCN‐134‐2D was systematically studied. The Tyndall scattering of PCN‐134‐2D showed a stable colloidal suspension . The Zr‐BTB sample before and after TCPP incorporation exhibits similar N2 adsorption isotherms and almost identical pore size distributions, suggesting that TCPP was attached to the surface of the Zr‐BTB layer without occupying the MOF cavities. The slightly reduced surface area and N2 total uptake is explained by the increased formula weight after TCPP incorporation.To investigate the distribution of TCPP within PCN‐134‐2D, the Ni‐porphyrin‐based ligand (TCPP‐Ni) was used for the synthesis of PCN‐134‐2D‐Ni. The Ni in the porphyrin center was intended to act as a probe indicating the position of the TCPP ligand using elemental mapping by SEM/energy‐dispersive X‐ray (EDX) analysis and TEM/EDX. Elemental mapping indicated an overlapped distribution of Zr and Ni, thus demonstrating that the TCPP linkers were evenly distributed throughout the Zr‐BTB layer . Further evidence for the distribution of the TCPP ligands were provided by the NFigure1H‐NMR, and elemental analysis, the composition of PCN‐134‐2D was determined to be [Zr6O4(OH)4](OH)4.2(H2O)4.2(BTB)2(H2TCPP)0.9.We propose that the TCPP was bridging a pair of adjacent Zr‐clusters within the PCN‐134‐2D Figure4a, sim2.51O2 generation because of their high light harvesting efficiency.1O2 has a variety of potential applications in organic synthesis, wastewater treatment, and photodynamic therapy.1O2 generation. To determine the efficiency of this process, we utilized a well‐known 1O2 scavenger, 1,3‐diphenylisobenzofuran (DPBF) to monitor the 1O2 generation by the porphyrin‐based MOF catalysts. Upon visible‐light irradiation (λ > 420 nm) in the presence of PCN‐134‐2D, the absorption at 410 nm decreased, indicating the formation of 1O2 ‐2D exhibits much higher yield of artemisinin, 64%, which has surpassed all reported photocatalysts for the semisynthesis of artemisinin  and their 3D counterparts (PCN‐134‐3D) were compared, highlighting the enhanced substrate diffusion and catalyst accessibility of the 2D MOFs. Significantly, PCN‐134(Ni)‐2D exhibits highest yield of artemisinin among all reported homo‐ or heterogeneous photocatalysts for the artemisinin production. Considering the structural diversity and functional tunability of 2D MOFs, a variety of 2D MOF catalysts can be envisioned from the results of this work. Of most important significance, 2D MOF catalysts will bring new concepts to catalysis by blurring the distinction between homogeneous and heterogeneous catalysts while still preserving the desirable attributes of both the systems.In conclusion, we have demonstrated that a 2D MOF, Zr‐BTB, can act as a suitable catalyst support. Surface modification of Zr‐BTB nanosheets with porphyrin ligands were shown to give rise to an efficient The authors declare no conflict of interest.SupplementaryClick here for additional data file.


Much attention has recently been devoted to finding ways to strengthen global governance mechanisms of AMR. The call for implementing binding global commitments and strengthening global institutions has figured prominently in this debate. Less attention has been paid to the tensions between global and local solutions. These tensions are particularly prominent in AMR stewardship, i.e., in the processes of formulating policies and strategies to deal with AMR. This commentary argues for an increased focus on glocal governance to strengthen AMR stewardship.Infectious Diseases that legally binding governance mechanisms are necessary for strong, swift, and coordinated action to curb AMR [Lancet emphasized the need for binding legal agreements and establishing a novel supranational entity to facilitate such agreements [Calls for increased global antimicrobial stewardship to curb antimicrobial resistance (AMR) have been heeded in recent years. Many have pushed for legally binding international agreements and the establishment of a supranational entity dedicated to curbing AMR. The director of the WHO’s AMR Secretariat argued in a co-authored article in curb AMR . A multireements . Severalreements . Most rereements .Interestingly, all these calls included comparisons to climate change. Indeed, the similarities between AMR and climate change are striking: both have adverse consequences today that might potentially turn catastrophic in the future; both risk tragedies of the commons where the benefits from antibiotics and emissions are local but the costs of resistance and climate change are global; both contain clear ethical dilemmas where some overuse a common resource  while others lack access to the resource; and responding to both threats includes high complexity and multiple constituencies.However, one key lesson to take away from the international efforts to curtail climate change is that binding commitments are not necessarily effective. Several key states opted out of these climate treaties. Even the most prominent global governance effort, the 1997 Kyoto protocol, had only 37 industrialized countries committed to legally binding reductions in emissions, and they have mixed results to show for it . Legallyglocalization can provide insights on how to establish such more complex global governance regime.The global challenge of AMR, more so than with climate change, requires solutions that are compatible with local needs and contexts. The equivalent to universal emissions targets is hard to find in the realm of AMR. In some countries, the most effective solution may be to provide incentives for patients, doctors, and farmers to reduce their use of antibiotics. In other countries, access to clean drinking water or fast diagnostics matter more than adjustments in antibiotic consumers’ behavior. This marked variability in both the state of public health and type of health challenge necessitates a broader pallet of solutions than the binary climate change option of either restricting the consumption of energy or lowering emission intensity. Hence, a combination of non-binding and binding agreements at multiple levels and across different sectors are more likely to be effective than the more comprehensive, wide-scoped treaties that we know from climate change. The analytical lens of Glocal governance, therefore, suggests that global challenges should be addressed by the effective reconciliation of the local and the global. Three concrete strategies can help facilitate a strong glocal governance regime of AMR.Examining AMR stewardship through the prism of glocalization places an emphasis on tensions between the universal  and the particular  . GlocaliFirst, from the perspective of glocal governance, effective AMR stewardship might be achieved by policy diffusion where ‘frontrunner’ countries or institutions set best practices/ideas that spread across nations and are adapted to local contexts . While apolycentric governance in the form of horizontal collaboration on a subnational level [Second, an effective glocal governance strategy of AMR might rely on al level . A globaProblem-Driven Iterative Adaptation (PDIA) is a glocalization strategy that focuses on solving specific problems in particular local contexts rather than transplanting “best practice” solutions [Lastly, olutions . It reliThe WHO has produced several useful scenarios for stronger global governance of AMR . Glocal The global challenge of AMR requires solutions that are compatible with local needs and contexts. Glocal governance might constitute an effective international regime of antimicrobial stewardship. Three concrete strategies can help facilitate a strong glocal governance regime of AMR: (i) policy diffusion where agenda-setting at the global level stimulates (but does not dictate) local policy innovation and practices; (ii) polycentric governance where subnational agencies form horizontal networks to solve particular problems; and (iii) problem-driven iterative adaptation practices that activate multiple levels of governance and sectors in addressing country-specific problems related to AMR.


Panax ginseng Meyer. In the present study, the potential therapeutic effect of G-F1 were investigated by computational target fishing approaches including ADMET prediction, biological activity prediction from chemical structure, molecular docking, and molecular dynamics methods. Results were suggested to express the biological activity of G-F1 against p38 MAP kinase protein. The p38 MAP kinase protein is an important signal transducing enzyme involved in many cellular regulations, including signaling pathways, pain and inflammation. Numerous studies are shown that an abnormal activation of p38 MAP kinase leads to variety of diseases. The pharmacokinetic result proves that G- F1 can act non-toxic drug like molecule. In addition, molecular level interaction results of G- F1 with p38 MAP kinase active (binding) sites residues clearly defines its inhibitory action on p38 MAP kinase. Further, molecular dynamics study also supported p38 MAP kinase and G-F1 structural stability. Findings from out study will assist to discover the active drug like molecules from Panax ginseng with help of molecular modeling techniques.Ginsenoside F1 (G-F1) is biologically an active compoud isolated from Korean  In pathological condition of PD there is successive reduction of dopamine (DA) supply levels in the striatum that causes imbalance between neurotransmitters like acetylcholine and DA as well as degeneration of non-dopaminergic mechanism such as cholinergic, noradrenergic and serotonergic systems in PD person ,2. Proba in-vivo . Dopamin in-vivo . In rece in-vivo , 6. It h in-vivo , 8. Seve in-vivo . It has  in-vivo -12. in-vivo  and in-vitro (SH-SY5Y and PC12 cells) models might assist as valuable measurement tools for the evaluation and efficacy for development of novel therapeutic agents for treatments of PD. Widespread study of these in-vitro and in-vivo models have provided significant cellular agents of cell apoptosis comprising excitotoxicity, mitochondrial dysfunction, neuro-inflammation, nitric oxide and oxidative stress  is used to treat human disorder as an oriental medicine. P. ginseng has three types of saponins commonly referred to as ginsenosides: protopanaxadiol (PPD), protopanaxatriol (PPT), and oleanolic acid  of G-F1. The primary aims of this study were to determine the biological activities of G- F1 and its molecular interactions with p38 MAPK.Various therapeutic agents are used for enhancements in dopaminergic remedies and the development of non-dopaminergic medications to treat PD, however, they are associated with side effects. In this regard, natural compounds are potent sources for treatment and management of PD. Currently the root of lic acid . Many stlic acid . It has lic acid , 18. Reclic acid , 20. In lic acid . Ginseno extract , 23. As Materials and MethodsPreparation of Protein and ligand moleculesP. ginseng saponin database. Its two-dimensional (2D) structure was drawn using ACD/ChemSketch (http://www.acdlabs.com) and converted to a three-dimensional (3D) structure by the OpenBabel program  online server (http://www.pharmaexpert.ru/passonline/) , which pMolecular docking simulationMolecular docking studies were performed with Autodock 4.2 -34 usingMolecular dynamics simulations+ or Cl-), followed by canonical ensemble and isothermal-isobaric ensemble equilibration. Individual systems underwent 25-nanosecond production. The root mean square deviation (RMSD) and root mean square fluctuation (RMSF) values were calculated using the Gromos program utilities g_rms and g_rmsf. All molecular dynamics simulations were performed using the Intel® 2.93 GHz Xenon® CPU 5679 RHEL 6 server. Molecular dynamics simulations were performed with a Gromacs96 43a2 force field  and a siPanax ginseng, plays important role in the pharmacological action of ginseng. Modern studies demonstrate that these ginsenosides are distinctive triterpenoid dammarane saponins and have a valuable beneficial effect on the protection and stimulation for CNS-related diseased conditions, mainly PD. Though, these ginsenosides are being used for decades, however, their pharmacological action and molecular level activity is still unknown. P38, one of the most probable the therapeutic target of PD, and the tested ginsenoside (G-F1) for neuroprotective behavior was selected for the current study. We examined the molecular interaction using in silico molecular docking to reveal the protein-ligand (p38-G-F1) interaction. The strength of the protein-ligand (p38-G-F1) interaction were determined by the binding free energy or docking score of the protein–ligand complex structure.Parkinson’s disease is the most common neurodegenerative disease that effects thousands of people throughout the world. During Parkinson’s disease, there is successive neurodegeneration and loss of neurons in the brain. Recently, it has been suggested that p38MAPK plays a significant role in hippocampal neurons apoptosis and cerebellar granule neurons, –46. TherScreening pharmacological properties (ADMET properties)In silico predictions have been used to determine ADMET descriptors such as serum protein binding capacity, blood-brain barrier crossing, central nervous system activity, and HERG K+ channel activity, apparent Caco-2, apparent MDCK, and skin permeability, percentage of human gastrointestinal absorption, hepatotoxicity, and CYP2D6 probability. We used Lipinski’s rule of five to determine the drug-likeness of G-F1. By this rule, most orally administered drugs have molecular weight less than 500, a distribution ratio less than 5, less than 5 hydrogen bond donors, and less than 10 hydrogen bond acceptors. G-F1 had a molecular weight of 638.88 Kda, a distribution ratio of 2.482, 7 hydrogen bond donors and 14 hydrogen bond acceptors. Although G-F1 did not obey Lipinski’s rule of 5, these values fall within the accepted ranges of 95% of known drugs. Additionally, toxicity estimation is one of the most vital task in natural compound screening (47), thus we estimated hepatotoxicity descriptors (0 is non-toxic and 1 is toxic) for tested ginsenoside by using ADMET module accessible from DS 2.5. The detailed results of predicted ADMET values of G-F1 with acceptable range were shown in Molecular interaction studiesThe p38 MAPK crystal structure was used to perform docking simulation using the Auto Dock 4.2 program. In this study we used SB203580 as a control molecule, which is a well-known drug for the inactivation and inhibition of p38 MAPK via its specific active sites including Lys 53 and Met 109. Structure stability refinement using molecular dynamicsIn order to examine and refine the structural stability of p38 MAPK, three individual molecular dynamics simulations were performed on the apo form of p38 MAPK and its ligand complexes with SB203580 and G-F1. SummaryIn conclusion, the ADMET and PASS results suggest that G-F1 is a non-toxic, drug-like compound with potential for p38 MAPK inhibition. In particular, the strong interaction between G-F1 and p38 MAPK involved four hydrogen bonds with two active sites . Molecular dynamics simulation suggested that the specified change less obviously was observed at coupling G-F1, p38 MAPK active site residues. Importantly, the p38 MAPK-G-F1 binding orientation did not affect the enzyme’s structural stability. Taken together, our results suggest that G-F1 has a potentially therapeutic role by inhibiting p38 MAPK. Our study concludes that ginsenoside F1 will be a promising compound for the development of anti-Parkinson’s therapeutic agents.


Acinetobacter baumannii, Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus were 1.92 (95% CI 0.10–35.52), 1.25 (95% CI 0.08–20.29), 1.60 (95% CI 0.13–19.10) and 1.84 (95% CI 0.04–95.58), respectively. In the complete absence of AMR bacteria, this study estimated that annually, in Thailand, there would be 111 295 fewer AMR cases and 48 258 fewer deaths.Antimicrobial resistance is a major health threat worldwide as it brings about poorer treatment outcome and places economic burden to the society. This study aims to estimate the annual relative increased in inpatient mortality from antimicrobial resistant (AMR) nosocomial infections (NI) in Thailand. A retrospective cohort study was conducted at Ramathibodi Hospital, Bangkok, Thailand, over 2008–2012. Survival model was used to estimate the hazard ratio of mortality of AMR relative to those patients without resistance (non-AMR) after controlling for nine potential confounders. The majority of NI (73.80%) were caused by AMR bacteria over the study period. Patients in the AMR and non-AMR groups had similar baseline clinical characteristics. Relative to patients in the non-AMR group, the expected hazard ratios of mortality for patients in the AMR group with  The discovery of antimicrobials was one of the most important medical advancements crucial to global health . AntimicAcinetobacter baumannii, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa and Staphylococcus aureus [P < 0.10 from the univariate analysis [A retrospective cohort study was conducted at Ramathibodi Hospital, Bangkok, a 1400-bed university hospital in Thailand, between 1 January 2008 and 31 December 2012. The population of this study were patients admitted to critical areas, namely, medical intensive care unit (MICU), intermediate medical care unit (IMCU), surgical intensive care unit (SICU), cardiovascular–thoracic surgical intensive care unit, and coronary care unit (CCU) who subsequently contracted NI in either critical areas or other wards. The patients in this cohort were identified by the infection control unit of Ramathibodi Hospital using the surveillance definitions of the United States Centers for Disease Control and Prevention (US-CDC) . There ws aureus –11. The s aureus –11. A Chs aureus . The eveanalysis  were conanalysis ; see SupThe prevalence of AMR NI reported in published national surveillance  was usedThis study protocol has been reviewed and approved by the Committee on Human Rights Related to Research Involving Human Subjects of Faculty of Medicine Ramathibodi Hospital, Mahidol University. The Institutional Review Board number for this study is ID 10-56-41. Data were anonymous, kept confidential and not linked to individuals.At the time of study, the hospital had nine intensive care units covering medical, surgical and paediatric populations, as well as general inpatient care units. There were 10 infection control nurses and one physician running the infection prevention and control based on the US-CDC surveillance strategies. Infection prevention measures were mainly those recommended by the US-CDC, the Society for Healthcare Epidemiology of America and WHO. The infection prevention and control programme has been performing prospective NI surveillance focusing on ICU patients continuously for decades.vs. 58%) in the MICU, IMCU and CCU were observed. The median length of hospital stay for patients in the AMR group was longer than those in the non-AMR group. The patients in both groups were different in the distribution of CCI score. Fifty per cent of patients acquired UTI, 30% acquired PNI, 15% acquired BSI, while the remaining patients acquired multiple site of infections. Among 523 critical care inpatients with NI, 201/386 (52%) patients with AMR and 53/137 (39%) with sensitive infections died. A. baumannii, E. coli, K. pneumoniae and S. aureus had a greater proportion of AMR than non-AMR infections but P. aeruginosa had a smaller proportion of AMR than non-AMR infections.Study results were reported following STROBE guideline for an observational study . There wA. baumannii, E. coli, P. aeruginosa and S. aureus, respectively. The other covariates adjusting for the Cox model comprised of sex, admitted ward, CCI score, site of infection, number of NI episode and the interaction effects between CCI score and site of infection.Hazard ratios of inpatient mortality and 95% confidence bounds for covariates in the Cox proportional hazard regression model were reported in A. baumannii was the most common cause of death. About 60 000 cases with resistant A. baumannii infections accounted for almost 40 000 deaths in that year.The estimated nationwide annual relative increased in inpatient mortality from AMR NI in Thailand was reported in A. baumannii, E. coli, P. aeruginosa and S. aureus were 92%, 25%, 60% and 84% higher, respectively. Because resistant E. coli in our study was ESBL-producing organisms which can be treated by widely available carbapenem antimicrobials, mortality of infection caused by this bacteria remained low. Another explanation is that the most common site of E. coli infection was UTI, which generally is not significantly associated with fatal outcome. On the other hand, A. baumannii mostly causes infection of the respiratory system which tends to carry higher mortality background. Only a few antimicrobial agents such as colistin, a nephrotoxic agent and tigecycline, a bacteriostatic agent with unpromising efficacy in the literature, are available for treatment. As a result, mortality associated with A. baumannii infection in our study was high. There were several studies that reported the mortality chances of these bacterial infections in Thailand and worldwide. Lim et al. published in 2016 [A. baumannii, E. coli, P. aeruginosa and S. aureus as 70%, 19%, 65% and 44%, respectively. Yang et al. published in 2013 [A. baumannii in hospital-acquired PNI and ventilator-associated PNI were 43.8% and 31.3%, respectively. Huang et al. published in 2012 [A. baumannii bacteraemia was 1.03 (95% CI 0.48–2.20) relative to carbapenem-susceptible A. baumannii bacteraemia. Abernethy et al. published in 2015 [E. coli was 18.2% (95% CI 17.8–18.7%). Schreiber et al. published in 2011 [S. aureus PNI was 39% compared with non-bacteria. The meta-analysis of Cosgrove et al. published in 2003 [P < 0.001) of mortality associated with methicillin-resistant S. aureus bacteraemia compared with methicillin-susceptible S. aureus bacteraemia. Our study estimated higher expected hazard ratios of mortality for AMR NI relative to non-AMR NI than other studies. Overall, mortality of AMR NI in our study was not statistically significant different from those of non-AMR. It might be due to the fact that these patients were all critically ill which posed them at risk of dying. Furthermore, with relatively more available resources for treatment as compared with other non-teaching hospitals, mortality of AMR NI might be lowered as shown by some studies that effective antibiotics played an important role in improving outcomes of these infections [We found that almost three-quarters of all patients with NI acquired resistant bacterial infections over the study period. The number of cases with AMR NI in our study was consistent with findings in a recent study in Thailand . The dis in 2013 , a retro in 2012 , a retro in 2003 , enrollefections , 28.et al. published in 2016 [et al. in 2017 [et al. in 2016 [This study estimated that the annual relative increased in inpatient mortality in Thailand would be 48 258 cases out of 111 295 (over 50%) cases with AMR NI. The study of Lim  in 2016  found 38 in 2016  used a det al. in 2003 [There are four main study limitations comprising the selection of an appropriate comparator, the lack of adjustment for acute severity of illness parameters, the potential statistical challenges to the estimation and the lack of accounting for the competing risk of being discharged in the multivariate Cox proportional hazard regression model. First, we have taken the more conservative approach of assuming that resistant infections are replacing susceptible ones so that we used the susceptible ones as comparators. The majority of studies to date have compared outcomes in patients infected with the resistant strain of an organism with patients infected with the susceptible strain of the same organism . We did  in 2003  compared in 2003 . Second, in 2003 . The datAMR has become a major public health threat and is spreading worldwide. Infectious diseases are the second main cause of death in low- to middle-income countries. This study sought to estimate the relative increased in inpatient mortality from AMR NI in Thailand to non-AMR NI using a multivariate Cox proportional hazard regression model. The Cox modelling was controlled for nine potential confounders while the AMR group has an elevated risk of inpatient mortality by 32%, it is not significant. In the absence of AMR bacteria, this study estimated that annually there would be 111 295 fewer AMR cases and 48 258 fewer deaths. The findings will assist in informing policy decision makers in their enactment of Thailand's National Strategic Plan on Antimicrobial Resistance for 2017–2021.


The mesolimbic hypothesis has been a central dogma of schizophrenia for decades, positing that aberrant functioning of midbrain dopamine projections to limbic regions causes psychotic symptoms. Recently, however, advances in neuroimaging techniques have led to the unanticipated finding that dopaminergic dysfunction in schizophrenia is greatest within nigrostriatal pathways, implicating the dorsal striatum in the pathophysiology and calling into question the mesolimbic theory. At the same time our knowledge of striatal anatomy and function has progressed, suggesting new mechanisms via which striatal dysfunction may contribute to the symptoms of schizophrenia. This Review draws together these developments, to explore what they mean for our understanding of the pathophysiology, clinical manifestations, and treatment of the disorder. Techniques for characterising the mesostriatal dopamine system, both in humans and animal models, have advanced significantly over the past decade.In vivo imaging studies in schizophrenia patients demonstrate that dopaminergic dysfunction in schizophrenia is greatest in nigrostriatal as opposed to mesolimbic pathways.Better understanding of striatal structure and function has enhanced our insight into the neurobiological basis of psychotic symptoms.The role of other neurotransmitters in modulating striatal dopamine function merits further exploration, and modulating these neurotransmitter systems has potential to offer new therapeutic strategies. Schizophrenia is a syndrome consisting of positive symptoms , negative ones (including flattened affect and lack of motivation), and cognitive ones. Dysregulated dopaminergic modulation of striatal function is fundamental to many models that seek to explain the mechanisms underlying the symptoms of schizophrenia In this paper, we review how advances in neuroscientific methods have improved our understanding of striatal structure and function. We then examine the evidence regarding striatal dysfunction in schizophrenia, and discuss how recent findings suggest a re-evaluation of prior hypotheses may be required. Finally, we ask what these developments mean for our understanding of the clinical manifestations and treatment of the disorder.The striatum is an integral part of the corticobasal ganglia circuitry. Extensive work mapping its pathways, as summarised below, suggests that it acts as an integrative hub for information processing in the brain.limbic Recent methodological advances have further refined our understanding of corticostriatal architecture In addition to its widespread cortical connectivity, the striatum has extensive bidirectional connections to the midbrain A. The deResting state functional magnetic resonance imaging (fMRI)diffusion tensor imagingMany of these more recent findings on striatal connectivity have been demonstrated only in rodents to date, and interspecies differences must be borne in mind when seeking to draw parallels with primate and human anatomy B [14]. HIn summary, it appears that, in addition to well-established parallel pathways, there also exists, across species, a high degree of pathway crossing and information funnelling, with regions in the associative striatum acting as integrative hubs. Furthermore, there appear to be a variety of pathways allowing for bidirectional information transfer across the striatum.Recent advances have refined understanding of striatal neurochemistry. In this section, we consider these findings and their implications for mechanisms by which abnormalities of nondopaminergic neurotransmitter systems may contribute to dopaminergic dysfunction. We also discuss their relevance for developing new treatment approaches to normalise striatal dopamine function and possibly treat schizophrenia without requiring dopamine D2/3 receptor blockade.Most dopamine neurons have the potential to release GABA as a cotransmitter, and a smaller proportion corelease glutamate. This cotransmission varies across the striatum, and can moderate the reciprocal relationship between dopaminergic and cholinergic neurons Corticostriatal neurons synapse upon cholinergic interneurons, which in turn modulate dopamine neurons via nicotinic receptors situated on these dopamine neurons, thereby mediating the corticostriatal control of striatal dopamine release et al. demonstrated that the extrapyramidal side effects of antipsychotic medications may primarily result from the blockade of D2 receptors on cholinergic interneurons indirect pathway MSNs, both due to the direct effect on these neurons, and as a result of increased acetylcholine-mediated activation of M1 receptors on the MSNs, occurring as a result of D2 blockade on cholinergic interneurons –fMRI studies also provide evidence that striatal dopamine has a broader role than simply encoding reward prediction errors, and is associated with the function of cortical salience networks The activity of mesostriatal dopamine neurons has been shown to signify the discrepancy between expected and actual rewards, which has been termed the reward prediction error amphetamine sensitisation encourages habit formation, and it is the dorsal striatum that is implicated in the habitual responses to drug cues experienced by addicts Striatal function along a ventral–dorsal axis is also relevant to habit formation. During the learning of action–outcome pairings, performance is goal directed and sensitive to changes in outcome values. After extensive training, however, performance moves to a stimulus–response mode that is inflexible, and no longer responds to changes in outcome. This evolution of behaviour from contingency-dependent learning to habitual responding has been associated with a shift from ventral to dorsal striatal processing positive psychotic symptoms Early postmortem studies investigating striatal dopaminergic abnormalities produced inconsistent results. Initial studies measured concentrations of dopamine, and while some reported an association between schizophrenia and increased concentrations specifically in the More recently, no differences between patients and controls in terms of either density of dopaminergic terminals in vivo. Radiolabelling of the dopamine precursor l-dihydroxyphenylalanine enables the measurement of its uptake and conversion in dopamine neurons to give an index of dopamine synthesis capacity. Alternatively, imaging the competition between endogenous dopamine and radioligands specific to postsynaptic dopamine receptors can determine baseline levels of dopamine (following depletion of endogenous dopamine using compounds such as α-methylparatyrosine), as well as the magnitude of dopamine release .PET allows quantification of the dopamine system Meta-analysis of studies using these PET and single photon emission computed tomography (SPECT) techniques shows that there is a robust increase in striatal dopamine synthesis and release in psychosis Early PET and SPECT studies of presynaptic dopamine function could distinguish anatomical subdivisions of the striatum but lacked sufficient resolution to accurately distinguish between functional subdivisions. Accordingly, these studies reported outcomes for the whole striatum or anatomical subdivisions. However, the subsequent development of higher resolution PET scanners has enabled the assessment of striatal functional subdivisions as well. The first studies to report on these subdivisions in patients with schizophrenia found that the greatest differences in dopamine function were within the associative striatum, with differences in the limbic subdivision not reaching statistical significance clinical high risk (CHR) of psychosis also found the greatest abnormality to be in the associative striatum Studies investigating presynaptic dopamine function in individuals at In a multimodal study, frontal activation during a working memory task was shown to correlate inversely with associative striatum dopamine synthesis capacity in a CHR group in vivo neuroimaging studies have provided evidence that dopaminergic dysfunction in schizophrenia is greatest within dorsal, as opposed to ventral regions of the striatum.In summary, in contrast to the mesolimbic theory, An ongoing question is how the neurobiological abnormalities identified in patients translate to the diverse psychopathology with which they present. We now suggest several mechanisms whereby striatal dysfunction could contribute to the clinical manifestations of the disorder. It is important to note, however, that many of these proposals are speculative at this stage, and furthermore, that schizophrenia is a heterogeneous disorder and no single brain region or neurotransmitter is likely to be able to account for all symptoms in all patients.Theoretical models linking biological substrates to phenomenological experience in psychosis have frequently built upon the finding that mesostriatal dopamine signalling is involved in marking the salience of environmental stimuli Existing models have proposed that delusions then develop secondary to cognitive processes attempting to construct a coherent explanation for these unusual experiences. Historically, however, discussions regarding the phenomenology of delusions have emphasised form of thought over content. Delusional form is characterised by a certainty of conviction, that is accompanied by an inability to shift perspective, and an imperviousness to counterargument Another finding of relevance to theories of aberrant salience attribution in schizophrenia is the following one. A stimulus, even if initially lacking inherent salience, once paired with dopaminergic activity, maintains the ability to evoke dopaminergic activity over time The striatum, and specifically areas of the associative striatum, can be viewed as an integrative hub. Regions of the caudate receive inputs from nearly the entire cortex inappropriate affect. However, this has yet to be definitively tested.Given that the striatum performs an integrative role, functional disruption secondary to aberrant dopamine signalling may lead to the associative impairments observed in schizophrenia. Based on preclinical models, it has been proposed that the underlying pathophysiology in schizophrenia represents a combination of increased aberrant spontaneous phasic dopamine release, and a reduction in adaptive phasic release in response to relevant stimuli Regionally targeted dopamine signalling enables the precise selection of specific behaviours over others, and collaterals between direct and indirect pathway MSNs efference copy is an internally generated replica of an outgoing motor signal, that has the effect of dampening sensory perceptions occurring as a result of the motor act, encouraging it to be perceived as self-authored and avoiding attribution to an external agent. The possibility, therefore, that disruption of efference copy mechanisms could contribute to passivity phenomena has long been suggested An Efference copies accompanying internally generated motor cortex activity travel via pyramidal tract neurons to the dorsal striatum thought echo. Inner speech is in certain respects a motor act, in that it is thought to result from motor plans for speech that are subsequently aborted In addition to motor passivity phenomena, a similar mechanism may contribute to auditory hallucinations related to inner speech, such as Efference-copy mechanisms, however, are less likely to account for auditory hallucinations that are phenomenologically unrelated to inner speech, and a predictive coding framework, which is a more generalisable model, seems more relevant in this context. Predictive coding refers to the idea that the brain compares prior expectations with new sensory evidence, and uses the discrepancy between the two (the prediction error) to update its model of the world. The certainty regarding one’s prior expectation is described as the precision of that prediction. Both the extent of the difference between prior and sensory data, and the precision of these determine the magnitude of the prediction error It addition to the role that striatal dopamine signalling plays in the development of positive symptoms, several mechanisms have been suggested for its contribution to the cognitive and negative symptoms of the disorder as well.in vivo imaging evidence for a deficit in cortical dopamine transmission has emerged Cognitive impairments in schizophrenia have been suggested to result from cortical hypodomaminergia; an idea supported by the importance of cortical dopamine signalling for prefrontal related cognition in vivo studies showing that reduced connectivity of the associative striatum and substantia nigra is related to the severity of cognitive deficits in schizophrenia Striatal hyperdopaminergia could conceivably result in cognitive impairments either by disrupting signalling between the frontal cortex and associative striatum, or by potentially driving cortical dopamine dysregulation There is also evidence that striatal dysfunction may contribute directly to negative symptoms. Studies using probabilistic learning tasks have shown that negative symptoms in individuals with schizophrenia may be related to impaired reward-based learning In this section we consider the implications of the evidence discussed earlier for the development of new treatments for schizophrenia that are not D2 receptor blockers and that might address the striatal dopamine dysfunction seen in the disorder.As previously discussed, studies have shown that the magnitude of dopaminergic abnormalities in schizophrenia varies across the striatum, with the most marked dysfunction seen in the associative striatum. These findings suggest that anatomically selective modulation of dopamine function is preferable The existing mechanistic understanding of striatal circuitry points at some possible therapeutic approaches that could allow more anatomically precise modulation of striatal dopamine function. One such mechanism, as discussed earlier, is muscarinic modulation of striatal dopamine release. Preclinical studies have shown that M4 positive allosteric modulators (PAMs) act on striatal MSNs to specifically inhibit dorsal striatum dopamine release via endocannabinoid signalling in vivo imaging evidence consistently suggests that the major abnormality in dopamine function in schizophrenia is located within the dorsal rather than the limbic striatum. Increasing knowledge regarding the structure, function, and neurochemistry of the striatum has improved our understanding of how these dopaminergic abnormalities may lead to symptoms. While these developments highlight potential pathways for the development of new treatments, the translation of these advances to meaningful clinical interventions, remains a significant challenge (See Outstanding Questions).Outstanding QuestionsIs anatomically precise modulation of dopamine signalling within the striatum possible? The variation across the striatum both in terms of dopamine receptor distributions, and of the mechanisms that control striatal dopamine and striatal function, suggests that it may be, but this remains to be tested.Do endocannabinoids, GABAgeric, cholinergic, and glutamatergic interventions have therapeutic potential given their role in modulating striatal function?Do primary striatal abnormalities exist in schizophrenia, or is dysfunction entirely secondary to upstream pathology? Genes associated with schizophrenia, overlap significantly with genes expressed by MSNs but not with those expressed by dopamine neurons. Abnormalities such as patch-matrix differentiation, or ones associated with cholinergic interneurons might also contribute to striatal dysfunction in schizophrenia. Recently, greater densities of afferent excitatory synapses have been found in the NAcc of individuals with schizophrenia, this could, via the feedforward mechanisms discussed A, drive Are specific symptoms associated with dopamine dysregulation in specific striatal loci? For instance, are verbal hallucinations associated with dopaminergic abnormalities in striatal regions displaying connectivity to the secondary auditory cortex? Are motor symptoms associated with the motor striatum?What is the optimal striatal parcellation? Preclinical findings suggest current atlases may oversimplify the picture, and point at the importance of improving our understanding of striatal connectivity. Could we improve our ability to characterise striatal dopamine dysfunction by using individual participant functional or anatomical connectivity to parcellate the striatum in multimodal studies of schizophrenia?Recent RM declares no financial conflicts of interest. AA-D has received research support from Pierre Fabre, Otsuka, Forest, Pfizer, and Neurocrine; served on advisory boards of Roche, Otsuka, Lundbeck; and given lectures sponsored by Otsuka. She is an advisor and holds shares in System 1 Biosciences and in Storm Biosciences. ODH has received investigator-initiated research funding from and/or participated in advisory/speaker meetings organised by Astra–Zeneca, Autifony, BMS, Eli Lilly, Heptares, Jansenn, Lundbeck, Lyden-Delta, Otsuka, Servier, Sunovion, Rand and Roche. Neither Dr Howes nor his family have been employed by or have holdings/a financial stake in any biomedical company.


No difference was observed between placebo and GLP‐1 (9‐36)amide, or between placebo and exenatide, although exenatide had a similar potency as GLP‐1 (7‐36)amide for cAMP formation in vitro (EC50 of 0.093 and 0.054 nmol/L). However, exenatide reached maximum plasma concentration at 90 min versus 15 min for GLP‐1 (7‐36)amide, and plasma CTX was significantly decreased during the second hour of the study after exenatide injections compared with placebo . There was no effect of the injections on bone formation markers  or on OPG, PTH and IGF‐1 levels. In conclusion, we show that GLP‐1 receptor agonists, but not the primary metabolite GLP‐1 (9‐36)amide, decrease bone resorption, and suggest that GLP‐1 may be part of the gut–bone axis. © 2019 The Authors. JBMR Plus is published by Wiley Periodicals, Inc. on behalf of the American Society for Bone and Mineral Research.Bones have been suggested to be a target for glucagon‐like peptide ‐1 (GLP‐1); however, studies of the effects on human bones so far have given diverging results. We hypothesized that GLP‐1, together with glucagon‐like peptide‐2 and glucose‐dependent insulinotropic polypeptide, plays a role in the gut–bone axis. We examined the acute effect of three GLP‐1 receptor ligands  on markers of bone remodeling. Eight healthy, normal‐weight participants, with a mean age of 24.3 years, were studied for 4 days in a double‐blinded, randomized clinical trial. Blood was collected before and after s.c. injection of GLP‐1 (7‐36)amide (1.5 nmol/kg), GLP‐1 (9‐36)amide (1.5 nmol/kg), exenatide (2.4 nmol/subject), or saline. Plasma was analyzed for bone markers and for osteoprotegerin (OPG), PTH, and IGF‐1 levels. All ligands were tested in vitro for their cAMP‐inducing activity on the human GLP‐1 receptor. GLP‐1 (7‐36)amide decreased CTX‐levels, compared with placebo (area under the curve [AUC] ±SD 0 to 120 min = –2143 ± 1294 % × min versus –883 ± 1557 % × min;  Glucagon‐like peptide‐1 (GLP‐1) is a peptide hormone secreted from enteroendocrine L‐cells upon meal ingestion.GLP‐1 is primarily known as an insulinotropic hormone, which together with glucose‐dependent insulinotropic polypeptide (GIP), is responsible for the incretin effect, ie, the markedly increased insulin secretion after oral as opposed to i.v.‐administered glucose.The metabolite, GLP‐1 9‐36)amide, has little effect on insulin secretion or glucose metabolism in humans.amide, haExtrapancreatic effects of GLP‐1 7‐36)amide include the inhibition of appetite, food intake, and gastrointestinal motility6amide inBone remodeling shows a diurnal variation, in particular, a decrease in bone resorption is observed postprandially, which can be eliminated by fasting.A study in ovariectomized mice showed that GLP‐1 administration was associated with an increase in BMD, decreases in RANKL expression, and increases in the bone formation markers, osteocalcin (OC), and P1NP secretion.GLP‐1 receptors have been observed on premature human osteoblasts, but not on mature human osteoblasts.The long‐acting GLP‐1RA, liraglutide, has been reported to promote the differentiation of both human and rat bone marrow stromal cells into osteoblasts and to inhibit adipocytogenesis.In vitro studies show that GLP‐1 may increase the number of osteoclasts, but that their resorptive area decreases, resulting in no effect on the resorption rate.Diverging effects of GLP‐1 have been found in humans. Liraglutide treatment was associated with a reduced risk of fractures, whereas exenatide was associated with an increased risk of fractures as compared with placebo or other antidiabetic drugs.To elucidate the possible acute effect of GLP‐1RA on bone remodeling, we examined the acute effect of s.c. injections of intact GLP‐1 (7‐36)amide, as well as the metabolite GLP‐1 (9‐36)amide and the degradation resistant GLP‐1RA, exenatide, on bone remodeling.2. All participants were normotensive and routine blood chemistry analyses showed normal values of creatinine, Na+, K+, CRP, leucocytes, alanine transaminase, aspartate aminotransferase, alkaline phosphatase, amylase, hemoglobin, glucose, HbA1c, and cholesterol. Data regarding the circulatory effects of GLP‐1 (7‐36)amide, GLP‐1(9‐36)amide, and exenatide, particularly with respect to mesenteric blood flow, were recently published.The study population consisted of eight volunteers , with a mean age of 24.3 years  and a mean BMI of 22.6 (SD 2.1) kg/mThe study was conducted as a double‐blinded randomized clinical trial. GLP‐1 (7‐36)amide, GLP‐1 (9‐36)amide, exenatide (exendin‐4), and saline were injected on 4 different study days under identical circumstances. Synthetic human GLP‐1 (9‐36)amide  as well as synthetic human GLP‐1 (7‐36)amide  were both dissolved in saline with 2% human serum albumin , sterile filtered, and dispensed into glass ampules that were afterwards tested for sterility and pyrogenes. The purity and structure of the peptides were examined by sequence, HPLC, and mass analysis. Exenatide  and isotonic saline are commercially available and were used without modification.The participants arrived after an overnight fast; the female participants underwent a urine hCG‐test . The participants were placed in the supine position and an i.v. access was established. Following a resting period (15 to 20 min), subjects received on the 4 study days and in random order: s.c. injections (1 mL) of isotonic saline, 2.4 nmol exenatide, 1.5 nmol/kg synthetic GLP‐1 (9‐36)amide, or 1.5 nmol/kg synthetic GLP‐1 (7‐36)amide.°C until time of analysis. Serum was stored in cryotubes at –80°C.Venous blood samples were collected in chilled EDTA tubes at time points –15, 0, 15, 30, 45, 60, 90, and 120 min, and centrifuged at 3600 rpm for 10 min at 4°C. Next, plasma was transferred to ice‐chilled tubes  and stored at –20Radioimmunoassays were used for measuring plasma concentrations of both GLP‐1  and exenatide after ethanol extraction, as described previously.Plasma insulin and C‐peptide were measured using a Cobas 8000, e602 module , using the following Cobas products: insulin reagents, insulin calibrator, C‐peptide reagents, and C‐peptide calibrator. The Cobas module uses a sandwich electro‐chemiluminescense immunoassay.Plasma samples were analyzed for OC, PTH, IGF‐1, CTX, and P1NP on an ids‐iSYS Multi‐ Discipline Automated System  by the automated chemiluminescence immunoassay method. The method is FDA‐cleared and CE marked. Osteoprotegerin (OPG) was measured using a commercially available sandwich ELISA kit . The ELISA uses specific monoclonal antibodies towards human OPG.2 and 37°C in Dulbecco's modified Eagles medium 1885 supplemented with 10% FBS, 2 mmol/L glutamine, 180 units/mL penicillin, and 45 g/mL streptomycin. Transient transfection of the human GLP‐1 receptor was performed using the calcium phosphate precipitation method.COS‐7 cells were cultured at 10% COy = 100% as baseline. Differences in AUCs and plasma levels of the hormones were examined by one‐way ANOVA for repeated measurements, followed by Dunnett's multiple comparison test. Differences resulting in a p value <0.05 were considered significant. Calculations and graphs were all made in GraphPad Prism 5 (GraphPad Software).Based on previous studies,The study was approved by the Regional Committee on Biomedical Research (SJ‐339). The study was performed in concordance with the Helsinki II Declaration, as well as the Danish Data Protection Agency. All participants gave oral and written consent to participate. The study has been registered at clinicaltrials.gov, Protocol Registration number NCT01988545.Plasma concentrations of GLP‐1 (7‐36)amide, GLP‐1 (9‐36)amide, exenatide, glucose, insulin, and C‐peptide were presented previously.50 of 0.093 and 0.054 nmol/L, respectively, whereas GLP‐1(9‐36)amide acted as a low potency partial agonist with an estimated EC50 of 188 nmol/L , as no Emax was reached even at 1 µmol/L GLP‐1(9‐36)amide.In vitro, all three GLP‐1 ligands acted as agonists of the GLP‐1 receptor. GLP‐1 (7‐36)amide and exenatide acted as full agonists for cAMP formation with ECB). The GLP‐1 (7‐36)amide injections resulted in a gradual decrease of CTX levels reaching 74.4% (SD ±10.4%) of the fasting level at 120 min, with a significant difference in the AUC0‐120min (±SD) between GLP‐1 (7‐36)amide (–2143 ± 1294 % × min) and placebo . Exenatide injections resulted in plasma levels of CTX of 79.8% (SD ± 18.6%) at 120 min. When measured over the entire period, there was no significant difference in AUC between exenatide compared with the placebo day. However, calculating the AUC during the second hour of the study  revealed a significantly larger AUC (±SD) in the exenatide group compared with placebo . The partial agonist, GLP‐1 (9‐36)amide, did not result in any significant changes in CTX over the entire period in comparison to placebo as the CTX level reached 87.5% (SD ± 12.4%) of fasting level at 120 min for GLP‐1 (9‐36)amide injections compared with 87.5 % (SD ± 20.4%) upon saline injections.Injection of the two strong agonists resulted in a decrease in the bone resorption marker CTX Fig. B. The Gp = 0.99).Absolute baseline values of CTX were 0.63 ± 0.33, 0.6 ± 0.25, 0.64 ± 0.33, and 0.59 ± 0.31 ng/mL on the study days with GLP‐1 (7‐36)amide, GLP‐1 (9‐36)amide, exenatide, and placebo, respectively (one way ANOVA A), and none of the injections led to significant changes compared with placebo.The bone formation marker P1NP showed very little variation from the baseline level during the study Fig. A, and nB; however, there were no significant differences between any of the hormones and placebo. Moreover, we found no effect of any of the injections on OC, IGF‐1, and OPG (data not shown).PTH did show some variation from baseline as apparent from Fig. The present study found an acute decrease in CTX after injection of GLP‐1 (7‐36)amide, whereas the metabolite GLP‐1(9‐36)amide had no impact. In addition, exenatide injections resulted in a decrease in CTX, but only during the last half of the experiment. However, the s.c. injections of exenatide resulted in a much slower increase in plasma concentrations as compared with GLP‐1 (7‐36)amide with peak concentrations at 90 min compared with 15 min for GLP‐1(7‐36)amide. This slower increase is probably because of the much lower dose and the extended half‐life of exenatide compared with GLP‐1 (7‐36)amide. Thus, the decrease in CTX after exenatide closely followed the plasma profile and was significantly different from placebo only in the last half of the experiments (60 to 120 min).The effect of GLP‐1 (7‐36)amide on bone resorption found in this study contrasts with the results of a study by Christensen and colleagues, where GLP‐1 did not have any acute effect on bone resorption.The findings in the present study are also in opposition to a previous study by Henriksen and colleaguesWe also examined the GLP‐1 metabolite, GLP‐1 (9‐36)amide, and found no effect of this on any of the markers of bone remodeling. This is in concordance with the very low potency observed for GLP‐1 (9‐36)amide regarding GLP‐1 receptor activation and the plasma concentrations reached (approximately 300 pmol/L), which were probably too low for sufficient activation of the GLP‐1 receptor. Previous studies have likewise described low potency, and partial agonistic properties of GLP‐1 (9‐36)amide.Previous studies of postprandial bone remodeling show that oral glucose  can lead to a decrease in bone resorption as measured by CTX levels to as low as 50% of the fasting level (baseline level).n = 8). Nevertheless, the group was large enough to reveal significant effects of the two agonists, whereas there was no indication whatsoever of an effect of the GLP‐1 metabolite. Furthermore, it might have been beneficial to increase the duration of the observation period because the CTX levels did not return (increase) to fasting levels following the GLP‐1‐mediated decrease in bone resorption. However, in previous studies of longer duration the nadir of CTX was reached at 120 min after oral glucose intake.A strength of the present study is that it was done in healthy young people, with normal glucose tolerance, eliminating skewed results because of, ie, lowered GLP‐1 sensitivity (as seen in patients with type 2 diabetes).In conclusion, the present study shows an inhibitory effect of the two GLP‐1R agonists, GLP‐1 (7‐36)amide and exenatide, on bone resorption in humans. GLP‐1 may therefore contribute to the regulation of bone turnover as part of the gut–bone axis together with the more established gut hormone mediators, GIP and GLP‐2.The authors declare that they have nothing to disclose and no conflicts of interest associated with this manuscript.


FBXW7, HLA-A, MED1, NOTCH2, NOTCH3, and TP53 had higher mutation rates in the CSMD1-inactivated subgroups. The Interferon-gamma score and immune signature score were elevated in CSMD1-inactivated subgroups. We identified several CSMD1-related pathways, such as the phosphatidylinositol signaling system and inositol phosphate metabolism. Our study using three large and independent datasets suggests that CSMD1-related gene signatures are associated with the prognosis of HNSCC patients.Although the genetic alteration of CUB and Sushi multiple domains 1 (CSMD1) is known to be associated with poor prognosis in several cancers, there is a lack of clinical relevance in head and neck cancer. The aim of this study was to offer insight into the clinical significance of CSMD1, utilizing a multimodal approach that leverages publicly available independent genome-wide expression datasets. CSMD1-related genes were found and analyzed to examine the clinical significance of CSMD1 inactivation in the HNSCC cohort of publicly available databases. We analyzed the frequency of somatic mutations, clinicopathologic characteristics, association with immunotherapy-related gene signatures, and the pathways of gene signatures. We found 363 CSMD1-related genes. The prognosis of the CSMD1-inactivated subgroup was poor.  Head and neck squamous cell carcinoma (HNSCC) arises from mucosa lining the paranasal sinuses, nasal cavities, oral cavity, nasopharynx, oropharynx, hypopharynx, and larynx . DespiteCUB and Sushi multiple domains 1 (CSMD1) is a large ~390 kDa) membrane-bound complement inhibitor 90 kDa me. CSMD1 iThe CSMD1 gene occupies over 2 Mb in the short arm of chromosome 8 (8p23) . DeletioThe aims of this study were to find the gene signature related to CSMD1 and to investigate the clinical significance of CSMD1 utilizing a multimodal approach that leverages three large, publicly available, independent genome-wide expression datasets.p < 0.001 and Pearson correlation coefficient >0.4 or <−0.4). The expression of 363 genes was tightly associated with mRNA expression of CSMD1, and was selected as the CSMD1 signature (n = 513) revealed 2 distinct subgroups of HNSCC; CSMD1-activated  subgroup and CSMD1-inactivated  subgroup (p = 3.7 × 10−48). The Kaplan–Meier plots and the log-rank test showed significantly different overall survival (OS) between two subgroups. The OS at 5 years was 52% in the CA subgroup and 46% in the CI subgroup. The OS of patients in the CI subgroup were significantly worse than those of patients in CA subgroup . For this reason, we hypothesized that several related genes would play a role together rather than the sole role of CSMD1 in HNSCC. We sought to find genes involved in the inactivation of CSMD1. This study used datasets from three independent cohorts. ignature . Using tsubgroup A. The CAn = 270; Greece cohort, n = 109). The patients from the test cohorts were classified into CA and CI subgroups by prediction model based on BCCP. The Kaplan–Meier plots and the log-rank test showed significantly different prognosis between two subgroups. Kaplan–Meier plots and the log-rank test showed significant differences in OS in the Leipzig cohort , because of the available clinical data. In the univariate analysis, age (<60 years old vs. ≥60 years old), anatomic site (oropharynx vs. other site), T stage (T1 & T2 vs. T3 & T4) and the CSMD1 inactivation were significantly associated with overall survival. In the multivariate analysis, T stage and CSMD1 inactivation were independent prognostic factors in HNSCC   status, gender, smoking status, regional lymph node (LN) metastasis and T stage , Table 3p = 2.2 × 10−16). When comparing the HPV status in groups across CSMD1 signature, 81 of 321 patients in the CA subgroup (25.23%) were HPV (+) and 240 patients (74.76%) were HPV (−) status. Of 225 CI subgroup patients, 14 (6.22%) were HPV (+) and 211 (93.77%) were HPV (−). A clinical feature of the CI subgroup presented lower frequency of HPV (+) status than the CA subgroup (p = 1.57 × 10−8). When comparing gender in groups across CSMD1 signature, 77 of 431 patients of CA subgroup (17.86%) were female and 354 patients (82.13%) were male. Of 320 CI subgroup patients, 98 (30.62%) were female and 222 (69.31%) were male (p = 6.25 × 10−5). In addition, we also assessed association with smoking status and CSMD1 signature. 77 of the 424 CA subgroup patients (18.16%) were non-smokers and 347 patients (81.83%) were smokers. 81 of the 316 CI subgroup patients (25.63%) were non-smokers and 235 patients (74.36%) were smokers (p = 0.018). LN metastasis and T stage between CA and CI subgroup were not significantly different.Comparing tumor site, patients in CA subgroup were comprised of 35.83% oral cavity cancer, 30.21% oropharynx cancer, 27.63% larynx cancer and 6.32% hypopharynx cancer, while 69.06% of the CI subgroup were classified to oral cavity cancer (n = 493). We selected the most frequently mutated genes from previous study of TCGA. We evaluated the frequency of somatic mutations in 30 genes associated with HNSCC. Among these genes, FBXW7, HLA-A, MED1, NOTCH2, NOTCH3, and TP53 showed significantly higher mutation rates for patients in the CI than in the CA subgroup . We found that the IS scores were significantly higher in the CI than in the CA subgroup (p = 0.0022). The patients with CSMD1 inactivation had higher INFG score and IS scores, suggesting that some of these might contribute to the response of immune checkpoint inhibitors such as anti-CTLA-4.We sought to assess the association between CSMD1 inactivation and tumor immunity. We compared the interferon gamma (INFG) score and immune signature (IS) score between CA and CI. The IS scores were developed for identifying the responders to immunotherapy in a mouse model treated with anti-CTLA-4 antibodies. As shown in Four GO biological process term enrichment analyses demonstrated that CSMD1 signature genes were characterized by synapse assembly, response to estradiol, positive regulation of synapse assembly and intrinsic apoptotic signaling pathway in response to DNA damage. Also, KEGG pathway function enrichment analysis for genes in CSMD1 signature was performed. Five KEGG pathways were found to be enriched, including inflammatory mediator regulation of TRP channels, phosphatidylinositol signaling system, glycine, serine and threonine metabolism, inositol phosphate metabolism, and small cell lung cancer .HNSCC is the fourth most common gene alteration of CSMD1 in pancancer (17%), with a mutation of 5.2% and a deletion of 21% A. This sOur results revealed that CSMD1 inactivation was significantly associated with low OS in training sets. In the validation set, CSMD1 inactivation was significantly associated with shorter OS and DFS at 5 years. This agrees with other studies that have reported an association between loss of the 8p23 region or CSMD1 and poor survival and high stage disease in prostate, bladder and head and neck cancer patients . This suFBXW7, HLA-A, MED1, NOTCH2, NOTCH3, and TP53 mutation rates, which could be responsible for treatment-related heterogeneity. FBXW7 and TP53 are known to be tumor suppressors [HLA-A, MED1, NOTCH2 and NOTCH3 are related with cancer prognosis [In our study, CSMD1 inactivation was accompanied by higher pressors ,15. Alsorognosis ,17,18. ITherapies targeting the immune checkpoint molecules PD-1 and PD-L1 have achieved remarkable clinical responses in multiple types of cancers, including HNSCC. Identification of biomarkers that predict clinical benefit to immune-based approaches is needed. We sought to assess the association of CSMD1 inactivation with immune activation, such as elevation of INFG and IS scores. The analysis revealed that the CSMD1 inactivation subgroup had a potential response to immunotherapy. The IS scores reliably reported responders to immunotherapy in a mouse model treated with anti-CTLA-4 antibodies . Most imp = 0.021) and inositol phosphate metabolism (p = 0.028). Another important signaling pathway in cancer, including HNSCC, is the PI3K–PTEN–AKT pathway; the class Ia PI3Ks, which are most frequently associated with cancer, are heterodimers coupled to receptor tyrosine kinases such as EGFR or adaptor molecules that may become active after receptor phosphorylation [The biological processes and pathways of CSMD1 signature genes are shown in rylation . It has rylation . ITPKA, n = 513) was downloaded from the UCSC Cancer Genomics Browser (Available online: https://genome-cancer.ucsc.edu/). The data from the Institute for Medical Informatics, Statistics and Epidemiology  and AHEPA Hospital in Thessaloniki  were obtained from the National Center for Biotechnology Information (NCBI) Gene Expression Omnibus (GEO) database (Available online: http://www.ncbi.nlm.nih.gov/geo) and used as the test sets [All clinical and gene expression data were collected previously and are available from public databases. Gene expression data of The Cancer Genome Atlas  was used for analysis of gene expression data [p-value was less than 0.001 and the correlation coefficient was more than 0.4 or less than −0.4. Hierarchical clustering analysis was done using Cluster 3.0 program and a heatmap was generated using TreeView software programs [The BRB-ArrayTools software program  was adopted . Gene exThe clincopathologic characteristics, such as tumor sites, human papillomavirus (HPV) status, gender, smoking status, regional lymph node (LN) metastasis and T stage, were collected in the TCGA and Leipzig cohorts because of available data. These data were analyzed to assess the association with CSMD1 signature.The list of genes of CSMD1 signature was submitted to the DAVID bioinformatics resources 6.7 , to discover the gene ontology (GO) categories with significantly enriched gene numbers ,31. The CXCL9, CXCL10, IDO1, IFNG, HLA-DRA, and STAT1) was previously identified in melanoma samples from the KEYNOTE-001 study [A six-gene signature of INFG-related genes .To test the prognostic significance, only gene expression data with available survival data were used. Pearson correlation was used for correlation analysis. Standardization was obtained by subtracting the median of the gene from the value of each gene and dividing by the standard deviation. Kaplan–Meier plots were constructed and log-rank tests performed for comparing the survival. Independent Our study suggests that CSMD1 inactivation is associated with the poor prognosis of HNSCC patients. CSMD1 inactivation might be related to HPV and tumor immunity.


The rise of antimicrobial resistance is a global threat and Africa like any other developing setting is heavily affected. As one of its missions, the Africa CDC is poised to ensure this rising trend takes a diminishing route in the few years ahead. Diagnostics obviously play a pivotal role in AMR control and the advanced course in diagnostics (ACDx) has been instrumental in training critical decision makers over the past 7 years. This commentary presents an analysis of discussions and exchanges during the launch of the Africa CDC AMR surveillance network at the ACDX and the way forward for its implementation. The diagnostics priorities are also highlighted. There a century .The international response has been unprecedented and encouraging. While there are ongoing efforts towards the establishment of a global fund for AMR, the World Health Assembly (WHA) in May 2015 endorsed a global action plan to tackle AMR and has called on governments and private sector commitments towards mitigating the AMR crisis . Consequth session was unique because it was dedicated to AMR. Taking into account the pervasiveness of diagnostics in tackling AMR and that the ACDx is a forum where experts and policy makers from academia, industry, government and other private sector agencies converge, there was no better opportunity to launch the Africa CDC AMR Surveillance Network (AMRSNET). This diverse gathering was an essential starting point in the creation of sustainable public private partnerships that can drive and support the AMRSNET in fulfilling its goals and challenges. The objectives of the launch of the Africa CDC Antimicrobial resistance (AMR) surveillance network was to enable participants to learn about the Africa CDC AMR Surveillance Network and how the network should function based on lessons learnt from other global, regional and country AMR strategies and policies as well as to brainstorm on the way forward for the use of diagnostics to combat AMR. This commentary presents an analysis of some of the exchanges that took place during the launch of the Africa CDC AMRSNET and the way forward for its implementation, taking into the consideration some overarching diagnostic priorities.This year's 8In a roundtable discussion with AMR experts from five African countries, immediate actions were obtained which will be very instrumental in defining specific in-country targets in the national strategic plans. From this roundtable discussion, it was clear that not all countries are at the same level in the control of AMR. Africa CDC would need to take this into consideration when implementing its plan on AMR in the African continent. The findings from the roundtable and the perceived challenges are shown in This economic case was instrumental in convincing the government of Zimbabwe in engaging in the implementation and scale up of quality assured diagnostics. A similar economic case would be needed alongside a business model to ensure the adoption of the surveillance network by the government. Such a business model will be crucial for the sustainability of the AMR surveillance network in the country. Secondly, there is need to conduct a survey on the current state of antibiotics consumption in the continent. The Global Point Prevalence survey would be a good starting point. Global Point Prevalence Survey is the initiative of Professor Herman Goossens of the Institute of Tropical Medicine, Antwerp, Belgium. This survey, with funding from the European Council, is an innovative strategy to assess global antimicrobial consumption and resistance in hospital settings. It utilizes a simple and sustainable Web-based point system for instant report of hospital data on antimicrobial consumption . Data arHowever, the developers have confirmed that the system can be adapted with ease to capture the use of antibiotics in the veterinary, environmental and agricultural sectors. From conception, the developers were keen to ensure that the system should not be costly and should be done once to reduce the human resource constraints peculiar in low resource settings. In terms of the cost, it is estimated to cost about 120,000 Euros to run the system annually, taking into consideration the cost for the soft-ware and cost of payment of salaries of part-time staff. This system will likely be affordable and sustainable if implemented in Africa. The Africa CDC can emulate from the Global PSS example as follows: adapting the same system by recruiting more hospitals in Africa in order to have a more comprehensive picture of antimicrobial consumption in the continent. This should also involve adapting the use of the web based system to capture the veterinary and agricultural sectors in a One Health approach; review the costing to include human resource and logistics specificities in African health systems. It is estimated that for a 500 bed hospital, it would take just 2 weeks to capture all the surveillance data needed. This means even if veterinary and agricultural sectors are included, it would still be a cost effective and sustainable strategy for the AMRSNET to implement; with the availability of online E-modules for training on AMR, there will not be need to spend in organizing costly workshops on AMR. A major challenge for the AMRSNET is how to implement and monitor the progress in a phase based approach. From the deliberations by participants, it was suggested that lessons could be learnt from well-structured approaches such as SLIPTA . Based on the ISO 15189:2012(E) standard, the SLIPTA approach uses a checklist that outlines requirements for quality and competency aimed to develop and improve laboratory services towards accreditation .Laboratory mentors who have been trained in the implementation of the checklist are deployed and stationed in laboratories to provide technical support for the laboratories as well as to participate in the training of trainers. This approach has been scaled up in Africa with the support of the US CDC and the Africa Society for Laboratory Medicine (ASLM). A similar system could be adapted for the AMRSNET with the training of AMR mentors who can further train other AMR mentors in clinical, veterinary and agricultural sectors towards the step wise phase based approach. A scorecard that details targets to be met by the different sectors could be adapted for use in monitoring the implementation of this approach. Diagnostics play a crucial role in the control of infectious diseases and in the control of antimicrobial resistance. Researchers, policy makers and implementers really need to be clear on the priority diagnostic needs for antimicrobial resistance surveillance. From our consultations, the following could be considered as diagnostic priorities: the availability of a rapid test to distinguish bacterial and viral infection. The need to ensure the uptake of any of such diagnostics if available in future should be high among clinicians and health personnel.There are some ongoing efforts. For example, in Tanzania there is the use the e-POCT phone system with biomarkers such as CRP and pro-Calcitonin in reducing inappropriate antibiotic prescription in children. The results of this technology have been encouraging and should be encouraged in other African countries; there is need to select the diagnostic technologies that would be needed for surveillance. For example in Africa, if gram negative bacteria surveillance is chosen, major hospitals could be selected and affordable technologies that detect gram negative bacteria should be considered. If this is implemented, it would be easier to evaluate stewardship programmes aimed at reducing antimicrobial resistance; connectivity with current diagnostics used for AMR. This would help networking and real time implementation of action. Senegal declared a Dengue epidemic on October 24, 2017 and a lot can be learnt and leveraged from their current connectivity capabilities. Using a web based system, notifications about the epidemic can be obtained from cell phones, says Dr Amadou Sall of the Pasteur Institute in Dakar during an informal consultation at the ACDx. The Africa CDC can utilize the same connected system in its AMR network which will be able to be adapted to provide real time information of the strains of bacteria circulating in the network; continuous capacity building in diagnostics to support the AMRSNET. In conclusion, the launch of the Africa CDC AMRSNET is a very important step towards the implementation of its strategic plan on reducing the rate of antimicrobial resistance in Africa. However, lessons must be learnt from ongoing efforts so that we don't re-invent the wheel. Diagnostics certainly play a crucial role in the control of AMR, hence more attention should be given the urgent diagnostic priorities in the continent.The authors declare no competing interests.


A P2X1-eYFP knock-in mouse was generated to study receptor expression and mobility in smooth muscle and blood cells. eYFP was added to the C-terminus of the P2X1R and replaced the native P2X1R. Fluorescence corresponding to P2X1-eYFPR was detected in urinary bladder smooth muscle, platelets and megakaryocytes. ATP-evoked currents from wild type and P2X1-eYFP isolated urinary bladder smooth muscle cells had the same peak current amplitude and time-course showing that the eYFP addition had no obvious effect on properties. Fluorescence recovery after photobleaching (FRAP) in bladder smooth muscle cells demonstrated that surface P2X1Rs are mobile and their movement is reduced following cholesterol depletion. Compared to the platelet and megakaryocyte, P2X1-eYFP fluorescence was negligible in red blood cells and the majority of smaller marrow cells. The spatial pattern of P2X1-eYFP fluorescence in the megakaryocyte along with FRAP assessment of mobility suggested that P2X1Rs are expressed extensively throughout the membrane invagination system of this cell type. The current study highlights that the spatiotemporal properties of P2X1R expression can be monitored in real time in smooth muscle cells and megakaryocytes/platelets using the eYFP knock-in mouse model.The online version of this article (10.1007/s11302-019-09666-1) contains supplementary material, which is available to authorized users. ATP is co-released with classical neurotransmitters and acts at P2X1 receptors (P2X1Rs) to mediate neurogenic contraction of arteries, urinary bladder and the vas deferens . In addiATP opens the P2X1R channel resulting in an inward depolarizing current that decays during the continued presence of agonist (1–2 s) through a process called desensitisation. Receptor mobility has been suggested to contribute to the ability of a cell to respond to repeated applications of ATP . In ordeThe mouse P2X1R gene was linked in frame at the end of exon 12 to the enhanced yellow fluorescent protein (eYFP) via a 18 bp linker (5′-GATCCACCGGTCGCCACC-3′ corresponding to the amino acid sequence DPPVAT) in order to obtain the protein chimera P2X1-eYFP with the downstream neomycin cassette flanked by CRE-LoxP. Embryonic stem cells incorporating the P2X1-eYFP construct were selected by neomycin resistance and screened by Southern blot for correct homologous recombination of the P2X1R with P2X1-eYFP. Mice were then generated on a C57/Bl6 background and crossed with an MF1 CRE mice to remove the neomycin cassette (determined by PCR). Mice resulting from back crossing to MF1 of at least four generations were used in this study.2 and MgCl2 1 (pH adjusted to 7.3 with NaOH). Whole cell recordings were made with patch electrodes filled with a solution containing (in millimolar): Kgluconate 140, NaCl 5, HEPES 10, EGTA 9 (pH adjusted to 7.3 with KOH) using standard patch clamp recording conditions and ATP applied by U-tube as described previously : NaCl 150, KCl 2.5, HEPES 10, CaClP2X1-eYFP receptor distribution was imaged using a 60 × oil immersion 1.35 NA UPLSAPO objective lens on an Olympus IX81 microscope equipped with a FV1000 confocal laser scanning module . The dye was excited at 515 nm and the emission bandwidth was 530–630 nm. Receptor mobility was measured using fluorescence recovery after photobleach (FRAP)  and imagt tests were used for statistical analysis.Data are plotted as mean ± SEM and unpaired Primers that detected the P2X1-eYFP construct (and the absence of cre) were used to screen mice and select those that were either homo- or heterozygous for P2X1-eYFP (data not shown). Confirmation of the correct genotype was obtained by Western blotting Fig. . For wiln = 8,7). In addition, the time-course of current decay during the continued application was indistinguishable between WT and P2X1-eYFP expressing cells . This recovery is ~ 2-fold slower than that reported previously for the P2X1-GFP receptor expressed in HEK293 cells . For P2Fluorescence imaging also demonstrated robust expression of P2X1-eYFPRs in platelets Fig. . The plaP2X1-eYFP was also strongly expressed in megakaryocytes, with uniform fluorescence evident throughout the internal area with the exception of a region consistent with the polyploidic nucleus Fig. . FluoresThe present study shows that a P2X1-eYFP knock-in mouse is viable and can be used to map the receptor expression in real time. P2X1-eYFP fluorescence was detected in the smooth muscle of the bladder  as well as platelets and megakaryocytes. There was no obvious expression over background levels in the central nervous system. These results are consistent with previous reports , and comIn the present study, FRAP measurements demonstrate that P2X1Rs are highly mobile within the cell membrane in native tissues, as previously shown for expression systems . Such mo2+-dependent signalling events ESM 2(DOC 22 kb)