F = kx that describes AFM cantilever deflection, has not been dynamically controllable in the same way. Here we present and demonstrate a ‘stiffness clamp’ that can vary the apparent stiffness of an AFM cantilever. This method, employable on any AFM system by modifying feedback control of the cantilever, allows rapid and reversible tuning of the stiffness exposed to the sample in a way that can decouple the role of stiffness from force and deformation. We demonstrated the AFM stiffness clamp on two different samples: a contracting fibroblast cell and an expanding polyacrylamide hydrogel. We found that the fibroblast, a cell type that secretes and organizes the extracellular matrix, exhibited a rapid, sub-second change in traction rate (dF/dt) and contraction velocity (dx/dt) in response to step changes in stiffness between 1–100 nN/µm. This response was independent of the absolute contractile force and cell height, demonstrating that cells can react directly to changes in stiffness alone. In contrast, the hydrogel used in our experiment maintained a constant expansion velocity (dx/dt) over this range of stiffness, while the traction rate (dF/dt) changed with stiffness, showing that passive materials can also behave differently in different stiffness environments. The AFM stiffness clamp presented here, which is applicable to mechanical measurements on both biological and non-biological samples, may be used to investigate cellular mechanotransduction under a wide range of controlled mechanical boundary conditions.Atomic force microscopy (AFM) has become a powerful tool for measuring material properties in biology and imposing mechanical boundary conditions on samples from single molecules to cells and tissues. Constant force or constant height can be maintained in an AFM experiment through feedback control of cantilever deflection, known respectively as a ‘force clamp’ or ‘position clamp’. However, stiffness, the third variable in the Hookean relation  Atomic force microscopy (AFM), initially developed as a topographical imaging modality, has become an important tool for investigating the mechanical properties and dynamic behavior of biological molecules, materials, cells, and tissues The rigidity of the cellular microenvironment has been shown to be an important input signal that influences a range of biological processes The predominant methods for studying the effects of microenvironmental rigidity on cellular behaviors involve culturing cells on deformable substrates  proteins and other cells, can be modeled most simply as a cell pulling on a spring . Settingchange in force over the change in displacement and therefore cannot be controlled using conventional feedback routines. The AFM stiffness clamp presented here is able to dynamically tune apparent stiffness between the extremes of infinitely soft and stiff.In theory, a wide range of apparent stiffnesses may be achieved using only a single spring together with feedback control . If the ually is . If the We tested the ability of the AFM stiffness clamp algorithm to produce a range of apparent stiffnesses with an expanding hydrogel, and we characterized the material's response to step changes in stiffness. Addition of phosphate buffered saline (PBS) to a dehydrated ∼1 kPa polyacrylamide hydrogel caused it to gradually expand. As the gel expanded and increased in height, it pushed against the cantilever applying an increasing force . WithoutWith a single AFM cantilever with spring constant tiffness . The appFibroblast cells are used extensively as a model system to investigate the effect of substrate rigidity ntracted . Once coWe found that when the apparent stiffness changed to a larger value, the cell's traction rate ecreased . Notablyecreased  inset, iThe AFM stiffness clamp provides a high-resolution method for varying apparent stiffness and evaluating cellular responses including contraction behavior. Using the AFM stiffness clamp, we show that cells rapidly change their traction rate and contraction velocity in response to step changes in apparent stiffness. Importantly, the stiffness clamp algorithm dynamically changes the apparent stiffness while the force and height are unchanged in the instant before and after the stiffness change. Therefore, any cellular response is a function of the step change in stiffness and not force or height. This decoupling of stiffness from force and height unambiguously shows that stiffness changes alone caused the change in contraction.Our observation of stiffness dependent contraction of single cells is consistent with several previous studies. We recently used the high-resolution of AFM to characterize the contraction dynamics of single human platelet cells It is worthwhile to note that the AFM stiffness clamp presented here only alters stiffness in one axis, though as demonstrated above, this appears to be sufficient to elicit a response from the contracting cell. Due to the fact that stiffness can only be measured by displacing a sample, the apparent stiffness can only be applied when cell height is actively changing, for example during fibroblast contraction, cardiomyocyte beating, neutrophil shape change in response to chemoattractants, and cell rounding during mitosis.This AFM-based approach to dynamically tuning microenvironmental rigidity is broadly applicable to both biological and non-biological experimental situations. In essence, the algorithm we present can be applied to any system with a spring where there is precise knowledge of the force and a single means of adjusting the position of the spring base  experiments were conducted using a modified Veeco Bioscope I mounted on a Zeiss Axiovert 25 inverted microscope. The Bioscope I z-axis piezo in our system has a range of only 4 To monitor drift in both the cell and gel experiments, we placed the cantilever in contact with the glass substrate in force clamp mode, immediately before each experiment. Experiments were not started until the system had equilibrated, such that a force clamp could be maintained with no significant change in stage position . Drift over the course of the experiment was measured in two ways. First, the zero deflection point of the cantilever was compared before and after each experiment to measure any cantilever drift. Second, for cell experiments, the contact point between the surface and cantilever was measured before and after each experiment. These measurements confirmed that the drift over the course of the experiment was negligible compared to the active contraction of the cell and expansion of the gel. Drift accounted for <10% of the total deflection for all experiments used.The ∼1 kPa polyacrylamide hydrogel was dehydrated at 4°C overnight and was rehydrated immediately before the AFM experiment with a standard phosphate buffered saline (PBS) solution. The cantilever was brought into contact as the gel rehydrated and expanded.2 atmosphere. A trypsin solution was used to detach cells at which point trypsin neutralizer was added and cells were then centrifuged at 300 g for 5 minutes. The resulting supernatant was discarded and cells were resuspended in their culture medium (DMEM plus supplements). KOH cleaned glass substrates and cantilevers were immersed for 30 min in a 50 NIH 3T3 fibroblast cells were cultured in DMEM (GIBCO) supplemented with 10% fetal bovine serum, 100 U/ml penicillin, and 100 The inset of the


Hearing is especially important for most primate species as they live in habitats of dense vegetation that limits vision. Stebbins  summed uOur vision is limited not only in the dark but also outside the visual field. In fact, if we observe the behavior of a predator like a feline, oriented toward its prey, and at the same time a sound occurs behind, we might note three principal different behaviors: the predator could maintain its gaze and ears on the prey neglecting the sound source; the predator could maintain its gaze on the prey rotating ears and then shifting its auditory attention toward the sound source; finally the predator could break its attention and orient gaze and ears toward the sound source. A similar behavior is seen in human beings during social interaction with two or more interlocutors.In humans, orienting movements are carried out by the eyes, head, and/or body operating alone or in various combinations depending on the behavioral situation. However, in non-human primates, such as macaque monkeys, head orienting movements and, more generally, gaze-shift are accompanied by ear orienting movements, which allow the shifting of auditory attention toward a sound of interest  support a medio-lateral organization for both auditory and visual systems. In particular, the lateral portion, corresponding to Area 8A or Frontal Eye Field (FEF), could play a role in receiving visual and auditory information from a central part of the visual field and then in organizing gaze-shift motor commands toward it. Otherwise, the medial portion, corresponding to Area 8B or Premotor Ear-Eye Field (PEEF), could play a role in receiving principally auditory information from a peripheral region of the space and then in organizing gaze-shift motor commands toward it.The  is a functional field in the prefrontal cortex, located in the rostral bank of the arcuate sulcus of macaques. FEF participates in the transformation of visual signals into saccade motor commands Schall, . FEF is Otherwise, it is well known, that FEF receive also information from the auditory system, participating thus, in the transformation of auditory signals into motor commands. An interesting study show in fact as FEF, in human, is a multimodal area involved in ultra-rapid responses for auditory and visual stimuli (Kirchner et al., The auditory cortical system is constituted by two different streams, termed dorsal and ventral, projecting to the frontal cortex in non-human primates (Romanski et al., In accord to anatomical evidence, many neurons in FEF respond to auditory stimuli. FEF has an interesting medio-lateral gradient regarding both the auditory spatial selectivity and proportion of auditory neurons. In particular, Azuma and Suzuki  showed tRecently, area 8B, located medially to FEF, has been renamed as a new frontal field: PEEF (Lucchetti et al., Through this opinion article, in light of PEEF discovery (Lucchetti et al.,


Metallo beta lactamases (MBL) are enzymes that have wide spread of activity and they confer a high level of resistance to all β-lactams including carbapenem, except aztreonam . MBLs rePseudomonas and 76 Acinetobacter isolates were obtained from Bangabandhu Sheikh Mujib Medical University (BSMMU), Bangladesh Institute of Research and Rehabilitation for Diabetes, and Endocrine and Metabolic disorders (BIRDEM) hospitals of Dhaka city. A total of 53 and 29 IPM resistant Pseudomonas and Acinetobacter isolates respectively were selected. EDTA- IPM micro dilution minimum inhibitory concentration (EDTA-IPM micro dilution MIC) method detected MBL in 44 (83%) IPM resistant Pseudomonas and 19(65.5%) Acinetobacter isolates. DDST with CAZ-0.1M EDTA and CAZ-2-MPA detected MBL in 73.6% and 67.9% of IPM resistant Pseudomonas and 55.2% and 48.3% of Acinetobacter isolates respectively. The detection rate was 67.9% and 66.1% in Pseudomonas and 51.7% and 44.8% in Acinetobacter isolates by EDTA-IPM and IPM-2-MPA methods respectively. In comparison to DDST, DPT with 0.1M EDTA showed higher sensitivity (89.7%) and specificity (100%) for detection of MBL in Pseudomonas and Acinetobacter. Isolates were also tested for AmpC β lactamase by DPT using chelating agent - aminophenyl boronic acid (APB) and it detected AmpC β lactamase in 6.9% and 21.1% MBL positive Pseudomonas and Acinetobacter respectively.There are no standard methods for the detection of MBL production in gram negative organism in routine Microbiology practice. The present study was undertaken to evaluate the screening tests like double disk synergy test (DDST) and disk potentiation test (DPT) using ceftazidime (CAZ) and imipenem (IPM) disks with chelating agents like EDTA and 2-MPA. A total of 132 Pseudomonas and Acinetobacter are emerging in our country. Rapid detection of these MBLs is necessary to institute appropriate treatment and effective infection control measures. Simple screening test like DPT using CAZ/IPM with 0.1M EDTA can be introduced into the routine clinical laboratories for their early detection.MBL producing


As 2012 draws to a close, I am delighted to report to our readers that IJCH has successfully transformed itself into an Open Access and completely online journal, joining the family of journals published by the capable hands of our colleagues in Co-Action.Without fixed publication dates, manuscripts are now published online as soon as they are copyedited and laid out, greatly reducing the time lapse between submission and publication.nd highest quartile in the Medicine (Misc) category. Clearly, continuing improvement of the quality of the journal is our priority.The Thomson impact factor (IF) for 2011 has been released. Our IF is 1.060, a slight decline from 2010 (1.065). We ranked 78 out of 131 in the category of Public, Environmental and Occupational Health. Another competing IF is the SJR which uses different math and is based on Scopus data rather than the Web of Science. Our score is 0.049, which places us in the 2The year 2012 also witnessed important changes in the journal's parent organization, the International Association of Circumpolar Health Publishers (IACHP). It has merged with the International Network for Circumpolar Health Research (INCHR) to form a new, stronger organization called Circumpolar Health Research Network (CircHNet). The new network consists of both institutional and individual members, and in addition to publishing IJCH, is also engaged in educational activities and promotion of international collaboration in research.th International Congress of Circumpolar Health was convened in August, 2012 in Fairbanks, Alaska. As with past congresses, it was a major success, which shall be evident when its proceedings are published in our journal as a Special Issue in 2013.The 15An important change in 2013 will be the implementation of a 500 euro publication fee for articles that have been accepted for publication. All manuscripts submitted prior to January 1, 2013 and already under review or production will be exempt from this fee. Individuals with financial difficulty can apply to Co-Action for a waiver. Ability to pay the fee will in no way affect or influence the editorial decision process. We consider this an important step towards making the journal financially self-sustaining. Our fees are quite low by the standards of most online Open Access journals.We thank the former Editor Dr. Tiina Ikäheimo and assistant Riitta Aittamaa in our former editorial office in Oulu, Finland, for facilitating the transition and finalizing the closure of IACHP. The new board members of CircHNet have worked tirelessly to kick-start the new organization, under the leadership of our President, Professor Peter Bjerregaard of Copenhagen.We look forward to a new and exciting year in 2013. Let me wish all our readers a Merry Christmas and a Happy New Year.Kue YoungEditor-in-Chief


To date, these congresses have a 46 year tradition and have taken place in 13 countries:The papers in this special issue of ZooKeys were presented at the 16th International Congress of Myriapodology (16ICM) held in Olomouc, the Czech Republic on July 20–25, 2014. International Congresses of Myriapodology are scientific meetings of scientists, students and enthusiastic amateurs with specific interest in millipedes, centipedes, symphylans and pauropods, as well as velvet worms. Those are organised under the auspices of the Centre International de Myriapodologie (1ICM: 1968 – Paris, France2ICM: 1972 – Manchester, UK3ICM: 1975 – Hamburg, Germany4ICM: 1978 – Gargnano, Italy5ICM: 1981 – Radford, USA6ICM: 1984 – Amsterdam, The Netherlands7ICM: 1987 – Vittorio Veneto, Italy8ICM: 1990 – Innsbruck, Austria9ICM: 1993 – Paris, France10ICM: 1996 – Copenhagen, Denmark11ICM: 1999 – Bialowieza, Poland12ICM: 2002 – Mtunzini, South Africa13ICM: 2005 – Bergen, Norway14ICM: 2008 – Görlitz, Germany15ICM: 2011 – Brisbane, Australia16ICM: 2014 – Olomouc, Czech Republic16ICM was the first myriapodological congress to be held in the Czech Republic. The 84 participants and 18 accompanying persons came from 24 countries on five continents, making the 16th Congress a truly international and global meeting of myriapod specialists (Figs The organisers of the 16ICM are very grateful to the Palacký University, Olomouc, which provided us a pleasant area for all scientific as well as social parts of the Congress. We also thank the Biology Centre CAS and partial support from several unnamed colleagues who allowed the participation of young students at this Congress.Many thanks to Lyubomir Penev and the friendly staff at Pensoft Publishers for making the 16ICM papers freely available to a global online audience.


Background. Scarcity of grafts for kidney transplantation (KTX) caused an increased consideration of deceased donors with substantial risk factors. There is no agreement on which ones are detrimental for overall graft-survival. Therefore, we investigated in a nationwide multicentre study the impact of donor and recipient related risks known before KTX on graft-survival based on the original data used for allocation and graft acceptance. Methods. A nationwide deidentified multicenter study-database was created of data concerning kidneys donated and transplanted in Germany between 2006 and 2008 as provided by the national organ procurement organization (Deutsche Stiftung Organtransplantation) and BQS Institute. Multiple Cox regression  was conducted . Results. Risk factors associated with graft-survival were donor age (1.020 [1.013–1.027] per year), donor size (0.985 [0.977–0.993] per cm), donor's creatinine at admission (1.002 [1.001–1.004] per µmol/L), donor treatment with catecholamine (0.757 [0.635–0.901]), and reduced graft-quality at procurement (1.549 [1.217–1.973]), as well as recipient age (1.012 [1.003–1.021] per year), actual panel reactive antibodies (1.007 [1.002–1.011] per percent), retransplantation (1.850 [1.484–2.306]), recipient's cardiovascular comorbidity (1.436 [1.212–1.701]), and use of IL2-receptor antibodies for induction (0.741 [0.619–0.887]). Conclusion. Some donor characteristics persist to impact graft-survival  while the effect of others could be mitigated by elaborate donor-recipient match and care. Donor and recipient parameters interact and jointly influence overall graft-survival after kidney transplantation (KTX). The individual decision to use a graft or not is guided by the question whether it will be helpful for the allocated recipient taking into account persisting organ shortage. As a result, grafts from elderly donors or from donors with comorbidities are used increasingly. However there is no agreement which donor factors may or may not directly affect transplant outcome in correlation to recipient factors . German The present study uses data from two institutional databases. For quality assurance and patient safety reasons, data on transplantation as well as follow-up surveys were reported to the BQS Institute for Quality and Patient Safety from 2006 to 2008. In 2006 the German national organ procurement organization Deutsche Stiftung Organtransplantation (DSO) implemented a nationwide database for all donor-derived data collected prospectively and directly in the donor hospitals. These data on donor characterization were used for the allocation of kidney grafts via Eurotransplant. Merging the two databases of DSO and BQS into a completely deidentified research database allowed us to analyse the impact of donor characteristics on graft-survival after KTX. Further methodological details have been published elsewhere –8.The study was performed in accordance with the guidelines for Good Scientific and Good Epidemiological Practice of the German Society for Epidemiology (DGEPI 2008) . EthicalThe study population consists of 4411 deidentified records of kidney grafts donated from brain dead donors in Germany (DBD) and transplanted as isolated grafts in different German centers between 2006 and 2008. The study was restricted to the evaluation of graft-survival because in a worst case assumption both graft-failure and returning to dialysis or death of the recipient with functioning graft can be related to factors known before the event of KTX.P value below 0.20 in univariate analysis and existed before the event of KTX were considered initially. A P value below 0.05 in multivariate analysis was considered as significant (stepwise forward selection). All analyses were performed with IBM SPSS Statistics 19 .In a first step, the impact of single relevant donor and recipient risk factors on graft-survival was analysed by means of log-rank tests  and univariate Cox regression . In a second step, a multiple Cox regression model was developed to examine the joined impact of several risk factors on graft-survival. In this model, only risk factors which showed a For donor laboratory data after admission first, last, lowest, and peak value before procurement were analysed.Graft-quality was judged as good, medium, or poor according to the assessment of the surgeon at procurement based on visual inspection . This isn = 1364) due to allocation rules. Therefore this could not be considered in analysis.Percentages of peak and actual panel reactive antibodies (PRA) were calculated after screening with the complement dependent lymphocytotoxicity test . UnfortuGraft-survival-times were calculated from the data on postoperative hospital stay and follow-up examination. Mean graft-survival-time was 120.7 days for recipients whose kidney graft failed during the study period and 358.6 days for censored cases. In case of a recipient follow-up missing graft-survival was censored for the most recent data actually available. Due to legal rules follow-up had to be terminated at 3 years.http://dx.doi.org/10.1155/2015/307230).4411 kidneys were donated and transplanted in Germany between 2006 and 2008. 2085 were recovered from female and 2326 from male donors. 1634 kidneys were transplanted into female recipients and 2777 into male recipients. The gender match does not reveal any differences in graft-survival. Further details about donor and recipient characteristics are summarized in Tables Univariate analyses showed that graft-survival is significantly influenced by increasing donor age, limited donor size, atraumatic cause of donor death, preexisting arterial hypertension, and coronary heart disease or reactive antibody status against cytomegaly virus .Graft-survival is not compromised significantly by other donor characteristics  as well Some medications used during donor maintenance are of significant protective effect on graft-survival Tables  and e1 sGraft and preservation quality  is of significant impact on graft-survival. Parameters describing kidney anatomy prospectively at procurement such as side of the kidney (left or right), the number of arteries or veins, and length of the ureter and kind of preservation solution used are without significant influence on graft-survival as well as rescue allocation or local versus national exchange.Cold ischemia time (CIT) has a pivotal role for graft-survival, particularly in the group of donors above 65 years of age Figures -2. We anRecipient age is of significant impact on graft-survival Tables  and e2. If patients need re-KTX, the risk of graft loss increases significantly. This seems to be linked to the current degree of immunization against HLA-antigens as the Kaplan-Maier survival-plots are very similar in their trend Figures -5, mainl(1)n = 330): urine leakage, lymphocele,marginal effect on graft-survival ((2)n = 386): wound dehiscence, severe bleeding, wound infection, and other,compromising effect on graft-survival ((3)n = 65): arterial or venous thrombosis.disastrous effect on graft-survival margiMultiple Cox regression of factors known before the event of KTX shows  that graThis is the first national investigation for Germany which takes into account joint donor and recipient factors on a multicenter-level known before the event of KTX. In contrast to other studies, the analysed donor data are real-time data used for organ allocation and terminal decisions by recipient centers to realize KTX. The corresponding recipient data were collected for quality assurance reasons according to German law. To ensure data consistency, our study was limited to KTX performed in Germany with grafts recovered in Germany only.Coordinators and procurement-teams are familiar with the risks factors identified by donor characterization, donor evaluation, and graft assessment at procurement . AccordiIn multiple Cox regression analysis  increaseThe use of vasopressors or catecholamines does not compromise graft-survival, which underpins the findings of Schnuelle et al. . TherebyNo appropriate evidence exists on which renal function parameters should be used for the assessment of kidney function. In this population only in multiple analyses creatinine values measured initially at admission are significantly related to graft-survival. This must be interpreted with caution since potential donors are not in a stable status as it is required for assessment of kidney function directly or indirectly by creatinine measurement  with nonThe issue of donor age and ischemia time (CIT) deserves careful attention because in the whole population in 84.7% of the cases CIT did not exceed 18 hours. When using the limit of CIT below 12 hours as reference and analysing only donors younger than 65 years of age no impact on graft-survival was observed for cases with CIT between 12 and 18 hours while graft-survival was significantly decreased if CIT exceeded 18 hours . But wheDonor related factors such as those summarizing disease transmission risks , 33, desThe impact of CIT and other factors on delayed graft function (DGF) was not investigated further because DGF occurs after the event KTX. Therefore concerning graft-survival DGF is a risk connected through by the risks known before KTX and created in addition afterwards.Concerning recipient related risk factors, multiple Cox regression analysis  reveals Some preparations used for induction therapy are without significant impact on graft-survival  while others turned out to be beneficial  in our analysis. This underpins the guidelines , 15. HowPatients with chronic renal disease (CKD) suffer from comorbidities. While a previous history of arterial hypertension is without significant impact on graft-survival, as expected, recipients with diabetes, coronary heart disease, and/or peripheral vascular artery disease are at risk for decreased graft-survival. We could not detect a significant relation between graft-survival and the time spent on dialysis before KTX at an average time of 5.8 ± 3.3 years in this study while patients on the waiting list are managed according to the guidelines , 15.In this study posttransplantation variables are not considered in multiple analysis because we investigated the interaction of donor and recipient parameter known until the moment of decision making whether an allocated organ will be transplanted on the selected recipient or not. However, the data reported to the BQS Institute for reasons of quality assurance include data concerning the postoperative course which should be investigated separately. According to the national guidelines , 33 rescFor donors below 65 years of age zero HLA-B or zero HLA-DR broad antigen mismatches seem to be beneficial concerning graft-survival. Due to technical restrictions no data about HLA-mismatches became available for donors within ESP, which caused an inacceptable rate of missing values for further multivariable analysis.Intensive care units in donor hospitals are liable for their treatment protocols which are in compliance with national and international recommendations , 30, 34.A limitation of the study is the short follow-up period, but when implementing the concept of mandatory quality assurance in medicine prospectively by law it was decided to follow up recipients only for three years. In the future, such programs should include longer follow-up periods within a transplant registry as requested by others , 4. HowePreimplantation or zero-time biopsy data were not available since there is no national consensus on how to perform, standardize, and merge this information including the pragmatic recommendation to do this according to the Banff classification in line with all other transplant biopsies taken later on , 38. PreSince this was the first national study using all recipient and donor data as they existed right before KTX we did not include expanded donor criteria definitions or donor risk index calculations –23, 39. Beyond the crucial functional and morphological assessment of a graft before KTX only donor age limits graft-survival significantly as unchangeable risk factor besides graft assessment at procurement and estimation of irreversible impaired renal function of the donor. On the recipient side age, cardiovascular comorbidity and immunological status are significant risk factors which determine graft-survival. This has to be considered in the detailed and complete characterization of the donor organ and the elaborate recipient selection when discussing further how to handle the combination of donor and recipient related risk factors when caring for recipients with different probabilities of long-term survival.In the supplementary all data investigated are shown. They which were used for the Cox Regression model devlopement accoridng to the details outlined in the mehtods section. Table e1 summarizes all donor data, table e2 summarizes all recipient data.


The acknowledgments section for this mini-review article was accidentally omitted.This work was supported by Grants PI13/01604, PI10/01074 from National R+D+I  cofunded by Instituto Salud Carlos III (ISCIII) and the ERDF (MC). Marie Curie People Cofund Fellowship, Seventh Framework Programme of the European Commission grant 267248:DIATRAIN (AGG-V). MS is a recipient of an undergraduate grant from the University of Barcelona. MC is a recipient of a Miguel Servet contract . This work was carried out in part at the Esther Koplowitz Centre.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Cancers,Dear authors, reviewers, and readers of Cancers, for which I now serve as Editor-in-Chief. First, thanks are due to my predecessor and the founding Editor-in-Chief, Dr. Robert Weiss. Over the past six years, he has done an outstanding job in shaping the journal to ensure its place as an online journal addressing both clinical and basic science issues related to cancer research. Citations of the journal are now available in PubMed and full text is available in PubMed central. Articles published in Cancers are also indexed by Embase, Scopus, and other databases. Dr. Weiss has always acted with the utmost academic integrity, which makes him a hard act to follow. I am very aware of the responsibilities that the Editor-in-Chief role entails, and I approach my new role with excitement. It gives me great pleasure to welcome you to the latest issue of Under my leadership, the journal will continue its open access format, which will certainly evolve to ensure that the journal takes full advantage of the rapidly changing world of information and knowledge dissemination. The journal will continue to publish high-quality clinical, translational, and basic science research on cancer prevention, initiation, progression, and treatment, as well as other related topics. Peer review will remain a vital component of our assessment of submitted articles. Articles will be reviewed by editorial board members, who will pre-select high-quality and original articles before the articles are reviewed by at least two reviewers who are experts in the field of the research. This will ensure that all published articles are of the highest quality. The ultimate goal is to achieve high citation numbers and high impact for the journal.I am fortunate to be supported by a highly effective team in the editorial office. The current group of Associate Editors work incredibly hard in the assessment and processing of submitted articles. A group of new Associate Editors who are experts in various fields of cancer research will be added to the editorial board, which will provide guidance for the journal.As I assume the new role of Editor-in-Chief, I welcome input from our readers and contributors. Together with our outstanding team of editorial board members and highly effective editorial office team, I look forward to taking the journal to the next level.


Multiple Sclerosis International has retracted the article titled “Consensus Guidelines for CSF and Blood Biobanking for CNS Biomarker Studies” [ Neurology December 1, 2009; 73(22): 1914–1922. doi: 10.1212/WNL.0b013e3181c47cc2 [ Multiple Sclerosis International and the authors, but an arrangement with the American Academy of Neurology and its publisher, Wolters Kluwer, to republish these guidelines did not include permission to publish in this journal.Studies”  for dupl


PPL methylation was higher in ESCC tissues than in normal tissues. When human ESCC cell lines were treated with 5-aza-2′-deoxycytidine (5-aza-dC), a DNA-methyltransferase inhibitor, PPL transcription was induced. Human KYSE270 ESCC cells do not stratify under ordinary culture conditions and rarely produce desmosomes; however, the forced expression of PPL promoted cell stratification. PPL induction also promoted adhesion to extracellular matrix but delayed cell migration. The abundance of desmosome-like structures was greatly increased in PPL transfectant as determined by transmission electron microscopy. Very low expression of another desmosome protein EVPL in ESCC, even in PPL transfectant, also supported the significant role of PPL in desmosome formation and cell stratification. Our results first indicate that the downregulation of PPL mediated by DNA hypermethylation, which may play an important role in the loss of ESCC stratification and likely in metastatic phenotype.Periplakin (PPL), a member of the plakin family of proteins that localizes to desmosomes and intermediate filaments, is downregulated in human esophageal squamous cell carcinoma (ESCC). Little is known, however, about the molecular mechanism underlying the regulation of PPL expression and the contribution of PPL loss to the malignant property of the cancer is unclear. We demonstrated that PPL mRNA expression was significantly reduced in ESCC tissues compared with that in normal tissues. Therefore, we hypothesized that CpG hypermethylation is the cause of the downregulation of PPL. Bisulfite-pyrosequencing of 17 cases demonstrated that the frequency of  To transfer various types of food to the stomach, the esophageal mucosa requires resistance to mechanical stress. One of the mechanisms supporting such resistance is the stratified structure of the squamous epithelial cells, which have many spines that intermingle with each other through dense desmosomes. Periplakin (PPL) is a 195-kDa, membrane-associated protein and a member of the plakin protein family, which comprises desmoplakin, envoplakin (EVPL), plectin, and bullous pemphigoid antigen 1. Plakin family proteins connect cytoskeleton elements to form intercellular junction complexes such as desmosomes Despite the nearly ubiquitous presence of PPL in normal squamous cells, PPL-knockout mice do not have obvious abnormalities PPL in ESCC and found abnormal hypermethylation in the promoter region in concordance with decreased mRNA expression levels. We restored PPL expression in an ESCC cell line and found that PPL plays a role in squamous cell stratification.CpG islands are frequently hypermethylated in cancer, leading to transcriptional repression and the subsequent reduction or loss of gene function Paired ESCC and normal tissue specimens were obtained from 19 patients who had undergone esophagectomy or esophagogastrectomy with confirmed diagnosis of ESCC. The patients were randomly selected from those who underwent surgery from January 2013 to June 2014 at the National Center for Global Health and Medicine (NCGM). This study was approved by the research ethics committees of the NCGM (121), and informed consent was obtained from all the patients before the samples were collected. The characteristics of the samples used in each assay are summarized in Table4 cells/well and after 18 h, then treated with 5-aza-2′-deoxycytidine  for 72 h. For passage or to detach cells, 0.25% trypsin-EDTA  was used. Cell growth was determined by MTT assay following the vendor's protocol . For adhesion assay, cells were labeled with 3′-O-acetyl-2′,7′- bis(carboxyethyl)-4 or 5-carboxyfluorescein diacetoxymethyl ester , and suspended in Dulbecco's modified Eagle's medium with 0.1% bovine serum albumin at the density of 2 × 105/well of 24-well tissue culture plates coated with type I colloagen, human fibronectin or laminin . After 1 h incubation at 37°C and washing, images were captured with a fluorescent microscope. Adhered cell area was quantified using NIH ImageJ software. Cell mobility was investigated using the Cytoselect would-healing assay kit . Quantification was performed using NIH ImageJ software measuring the area covered with cells as % of whole captured square area .Human ESCC lines KYSE70, KYSE140, KYSE150, KYSE270, KYSE410, and KYSE510 PPL, and EVPL were Hs00160312_m1 and Hs00157430_m1, respectively.Total RNA was isolated from tissues using RNA Bee RNA Isolation Reagent . After the RNA was treated with DNase I, double-stranded cDNA was synthesized using the High-Capacity cDNA Reverse Transcription Kit . Quantitative PCR was performed using ABI TaqMan probes (Applied Biosystems) as described previously. Threshold cycle numbers were determined using the Sequence Detector software and transformed as described by the manufacturer, with glyceraldehyde-3-phosphate dehydrogenase as the calibrator gene. The TaqMan Gene-expression assay IDs used in this study for To assess the PPL methylation status, bisulfite-pyrosequencing was performed with PyroMark Gold Q24 reagents and a PyroMark Q24 pyrosequencing machine . The PCR primers used in this study were 5′-AGTTGATATTGGGAGTAGGTGTTA-3′ and 5′-CAAATTCCCTAAAAACCCCTCTTAA-3′. The primer used for pyrosequencing was 5′-GGGGTTTTAGAATATAGG-3′.Human HaloTag® expression vector  and human PPL HaloTag® ORF clone in pFN21A was purchased from Promega  and transfected into KYSE270 cells using lipofectamine LTX reagent . Stably transfected cells were isolated using a MoFlo™ XDP Cell Sorter .Formalin-fixed, paraffin-embedded sections of surgical specimens from patients with ESCC were deparaffinized and rehydrated. Antigen retrieval was performed using an autoclave in 10 mmol/L sodium citrate buffer. Sections were stained with hematoxylin and eosin (H&E) or anti-human PPL antibody . A diaminobenzidine staining procedure was performed using the ImmPACT™ DAB peroxidase substrate kit , and hematoxylin was used for counterstaining.4 cells/well) were cultured in a slide chamber , washed with phosphate-buffered saline (PBS), fixed with 4% paraformaldehyde for 15 min, and permeabilized with 0.1% triton X-100 for 5 min. The cells were washed with PBS, stained with anti-human PPL antibody  at a concentration of 3 μg/mL at 4°C overnight, washed again with PBS, and incubated with Alexa Fluor 488-labeld goat anti-rabbit antibody . F-actin was visualized with Alexa Fluor 633-labeld Phalloidin (Life Technologies) at the concentration of 5 unit/mL, for 20 min at room temperature. Vectashield mounting medium containing 4′,6-diamidino-2-phenylindole  was used for DNA staining. Images were captured using a confocal microscopy system . In some experiments, stained KYSE270 cells were also observed with fluorescent microscopy .KYSE270 cells  at 4°C and then postfixed with an aldehyde-OsO4 mixture containing 1.25% glutaraldehyde, 1% paraformaldehyde, 0.32% K3[Fe(CN)6], and 1% OsO4 in 30 mmol/L HEPES buffer (pH 7.4) for 1 h at room temperature. The fixed blocks were dehydrated and then embedded in Quetol 812 . The blocks were sectioned to a 70-nm thickness with an ultramicrotome . The ultrathin sections were contrasted with EM stainer (Nisshin EM) and examined with an electron microscope . Cells on culture dishes  were washed briefly with PBS and fixed with aldehyde-OsO4 mixture , and 1% OsO4 in 30 mmol/L HEPES buffer (pH 7.4)] for 1 h at room temperature as previously reported method t test using the Prism 6 statistical program . All tests were two-tailed, and P < 0.05 were considered significant. Correlations were tested with a two-tailed Pearson calculation.The data were expressed as the mean ± standard deviation, and the results were compared by paired or unpaired Student's P = 0.0009). In accordance with those results, the PPL mRNA levels determined by RT-PCR were significantly decreased in the cancer tissues compared with those in the normal mucosa  in the normal and tumor samples, respectively. A paired t-test indicated that the tumor DNA was significantly more methylated than that in the normal tissues  decreased to around 60%. Higher concentration of 5-aza-dC did not further decrease methylation levels. Similar to that in the ESCC tissue samples, the decreased DNA methylation was associated with the induction of PPL transcripts , the growth of the PPL transfectants was almost the same as that of the mock transfectants on day 3, after which neither cell type increased its density further (Fig.To further understand the significance of PPL defects in ESCC, we investigated the effect of forced PPL expression in the ESCC cell line KYSE270. We prepared a PPL-transfectant cell line and a mock-transfectant cell line. The PPL expression level in the PPL transfectants was around eightfold higher than that in the mock transfectants Fig.. Initialther Fig.. When thther Fig..Z axis (Fig.Z stacks of PPL-expressing cells clearly demonstrated a stratified structure (Fig.The appearances of the PPL-transfectant and mock-transfectant cultures were different. The original KYSE270 cells and the mock transfectants grew mostly in a spreading shape on the culture dishes, forming a monolayer sheet with no stratification Fig.. In contaxis Fig., which sture Fig.. We usedture Fig.. We furtture Fig., which dThe PPL transfectants adhered more to the culture plates than the mock transfectants did. When sparsely cultured cells were treated with trypsin and EDTA, most of the mock transfectants became detached within 10 min, whereas the PPL transfectants remained attached Fig.. We testPPL is one of the proteins that make up desmosomes, so we looked at the cell–cell adhesions in PPL transfectant further in TEM images Fig.. In the ESCC is an aggressive form of cancer and one of the most frequent types of cancer in the world and especially in Asian countries PPL alone did not affect desmosome formation in skin epithelial cells PPL alone to induce such a dramatic change in the formation of desmosome-like structures and cell adhesion. In the squamous cells of the skin or urinary tract, PPL colocalizes with EVPL, another plakin family protein. A previous report found that in the esophagus, the EVPL level was highest in the outermost layers, whereas PPL is expressed in all the suprabasal layers Desmosomes consist of many proteins, and a previous study reported that a lack of


Journal,1 the authors requested that some errors in the Tables 2 (p164) and 3 (p165) of the original paper should be corrected. The revised version of the Tables are following.About the paper by Sabanayagam C et al. published in the September 2007 issue of the Journal regrets the errors.The


Our in silico analysis suggested that the predicted epitopes, either GPKVSLIDTSSTITI or EWISIVPNFILVRNT, could be a better choice as universal vaccine component against NiV irrespective of different isolates which may elicit both humoral and cell-mediated immunity.This study aims to design epitope-based peptides for the utility of vaccine development by targeting glycoprotein G and envelope protein F of Nipah virus (NiV) that, respectively, facilitate attachment and fusion of NiV with host cells. Using various databases and tools, immune parameters of conserved sequence(s) from G and F proteins of different isolates of NiV were tested to predict probable epitope(s). Binding analyses of the peptides with MHC class-I and class-II molecules, epitope conservancy, population coverage, and linear B cell epitope prediction were analyzed. Predicted peptides interacted with seven or more MHC alleles and illustrated population coverage of more than 99% and 95%, for G and F proteins, respectively. The predicted class-I nonamers, SLIDTSSTI and EWISIVPNF, superimposed on the putative decameric B cell epitopes, were also identified as core sequences of the most probable class-II 15-mer peptides GPKVSLIDTSSTITI and EWISIVPNFILVRNT. These peptides were further validated for their binding to specific HLA alleles using  Nipah virus (NiV), first reported in Malaysia in 1998 mainly causing nonlethal respiratory disease in pigs, is a nonsegmented single stranded linear negative sense RNA pathogenic paramyxovirus virus . In humaUsing available knowledge on immunity to other paramyxoviruses –10, both+ T-cells) and class II (recognizes CD4+ T-cells) alleles [ in silico models [ in silico methods for predicting epitope-based peptides for the utility of vaccine development against the deadly Nipah virus.Secreted and surface proteins of a pathogen are mostly antigenic and responsible for pathogenicity . The env alleles , 15. Moro models , for preAn outline of the methodology undertaken for this study has been portrayed in http://www.uniprot.org) and NCBI protein database (http://www.ncbi.nlm.nih.gov/protein) in FASTA format. These sequences were deposited in the databases obtained from different parts of NiV endemic regions such as Faridpur, Manikgonj, and Rajbari districts of Bangladesh as well as Malaysia and India at different time. The habitats of isolates include pigs and humans.The sequences of glycoprotein G and fusion protein F of different isolates of NiV have been retrieved from uniprot (http://www.megasoftware.net). The CLUSTALW algorithm along with 1000 bootstrap value and other default parameters were used to fabricate the alignment. The sequences were analyzed with a view to recognize the immunologically pertinent regions that were achieved by predicting epitopic peptides. An amino acid stretch must be of a minimum length for being considered as an epitope that we are aiming to design. Due to representative length of peptide that binds to HLA molecules, nonamers were selected as the minimum length of the conserved sequences for the prediction of epitope-based peptide in this study.Retrieved sequences were subjected to multiple sequence alignment using MEGA 5.05 software package -based prediction tool in Immune Epitope Database (IEDB) was applied. All the available MHC alleles were selected and the peptide lengths were set at 9.0 before making prediction. The parameters for immunogenicity detection  were normalized on a scale of 0 to 1 and were given a weighted score to prioritize the parameters in order to determine the immunogenicity.The cytotoxic T lymphocyte (CTL) epitopes from the conserved peptides were predicted using the NetCTL 1.2 server available at  in vivo  which alhttp://tools.immuneepitope.org/mhcii/). In this case, SMM-align method [50 for the predicted binders within 250 nM.Conserved peptides were also tested for predicting epitopes that interact with MHC class II molecules by selecting all the alleles in IEDB MHC class II binding prediction tool (n method  was emplhttp://tools.immuneepitope.org/tools/population/iedb_input) by maintaining the default analyses parameters just as it is . For calculating the population coverage in IEDB, latest data from allelefrequencies.net database (2011 and onwards) were used, which is the most comprehensive one available because of its huge population datasets. For individual population coverage, the tool computes the followings: (i) apprehended population coverage; (ii) average number of hits by the epitopes/HLA combinations recognized by different ethnic groups or populations; and (iii) minimum number of hits by the epitopes/HLA combinations recognized by 90% of the population (PC90). These calculations were made on the basis of HLA genotypic frequencies assuming nonlinkage disequilibrium between HLA loci.Prediction of T cell epitope is not enough for becoming a good candidate peptide as vaccine. It should be taken into consideration that, along with identification of MHC allele-specific T cell epitope, the predicted epitope(s) should effectively cover human population. Predicted epitopes showed interaction with different MHC alleles. To find out the human population coverage of the individual epitopes, predicted epitopic sequences with the corresponding Class I HLA alleles were submitted to the population coverage analysis tool of IEDB (http://tools.immuneepitope.org/tools/conservancy/iedb_input) available in IEDB analysis resource. The conservancy level of each potential epitope was calculated by looking for identities in all 18 protein sequences of different strains retrieved from database.Epitope conservancy of the selected epitopes was tested using epitope conservancy tool (http://www.imtech.res.in/raghava/abcpred/). To do so, conserved sequences with ≥0.4 VaxiJen scores and exomembrane topology were applied in prediction server by setting cut-off value at 0.51 and the length of the epitopes was fixed as decamer. Overlapping sequences were also filtered. The nonamers which were significantly superimposed (≥7 amino acid overlaps) on putative B cell epitope (decamer peptides) were considered for interpretation.Linear B cell epitopes are of variable lengths of peptides from 2 to 85 compared to that of T cell epitopes. Linear B cell epitopes were predicted using ABCpred prediction server  . The cdsx-, y-, and z-axes, respectively, to allow the epitope to bind to the binding groove of the HLA-C 07∗02. The size was set at 20, 40, and 40 Å in the x, y, and z dimensions, respectively. For predicting binding of 15-mer epitopes in the binding groove of class II allele (HLA-DR1 or DRB1∗01:01), the grid/space box center was set at 9.292, 26.935, and 40.729 Å in the x-, y-, and z-axes, respectively (after converting DRB1∗01:01 and epitope files into PDBQT format). The size was set at 58, 22, and 38 Å in the x, y, and z dimensions, respectively.The AutoDOCK tool from the MGL software package (version 1.5.6) was employed for docking purpose , 28. BotAll the analyses were done at 1.00- Å spacing. The exhaustiveness parameter  was kept at 8.00, while the number of outputs was set at 10. These parameters were performed in AutoDOCK tool. The docking was conducted using AutoDOCK Vina program based on the above-mentioned parameters. All the output PDBQT files were converted in PDB format using OpenBabel (version 2.3.1) and visualized in PyMOL molecular Graphics system. The best output was selected on the basis of higher binding energy.The 3D structure of MHC class I H-2Kb molecule complexed with octapeptide PKB1 (“KVITFIDL”) was retrieved from Protein Data Bank Database (ID: IKJ3) and visualized using PyMOL Graphics. The octapeptide was excluded before applying the structure of H-2Kb for comparing the validated data obtained for predicted structure of HLA-C 07∗02.Also, to assess HLA-C 07∗02-epitope docking results, octapeptide PKB1 (“KVITFIDL”) was used as the control. This peptide was docked with HLAs, HLA-C 07∗02, and H-2Kb. The test epitope(s) and the control peptide were docked by setting similar parameters for each trial and successful binding of this peptide to these HLAs was demonstrated. Finally, H-2Kb - KVITFIDL docking result was used as control to compare with the test docking results of HLA-C 07∗02 complexed with selected epitopes. A comparative analysis of the best binding energy  and the arrangement of the test and control epitope at the binding groove of MHC allele HLA-C 07∗02 was also performed.Crystal structure of HLA-DRB1 complexed with an endogenous peptide (“GSDWRFLRGYHQYA”) was retrieved from protein databank (PDB ID: 1AQD). The peptide was excluded from the structure and its binding to the binding groove of HLA-DRB1 was used as the control model to compare the binding models of the predicted epitopes “SEWISIVPNFILVRN” and “VFYQASFSWDTMIKF” using AutoDOCK Vina.A total of 18 sequences of G and F proteins of different isolates of NiV have been retrieved. CLUSTALW programme in MEGA software generated several conserved sequences with varying lengths. A total of 15 and 8 conserved sequences were found in G and F proteins of NiV, respectively. Conserved sequences generated in this method have been presented in Analysis revealed that 9 and 5 conserved sequences, respectively, from G and F proteins met the criteria of default threshold level, ≥0.4, in VaxiJen . On the T cell epitopes are processed peptides which could be recognized by the T cell receptors presented through Class I/Class II molecules on antigen presenting cells. VaxiJen analyses were used for the identification and reevaluation of T cell epitopes because some T cell epitopes tend to lose the antigenicity when reanalyzed. So, to increase the confidence level of prediction about the epitopes presented as a good T cell based epitope, these were cross-checked with the VaxiJen scores.50 value <250 nM. Using the same IC50 value in case of conserved sequences of F protein, 37 different MHC class I molecules showed binding interaction with 150 T cell epitopes ranging from two to as many as ten MHC class I alleles. Next step analyses were proceeded with the peptides that showed interaction with ≥5 MHC class I alleles and had VaxiJen score ≥0.5. A total of 26 nonamers from F protein and 12 nonamers from G protein fulfilled these criteria which have been presented in NetCTL prediction tool covering all supertypes created a total of 146 and 151 nonamers from the conserved sequences of G and F proteins, respectively. Analysis in SMM based IEDB MHC I prediction tool retrieved 59 T cell epitopes (peptides from G protein) that interacted with 29 possible MHC I alleles with the IC50) threshold of 500 nM identifies peptide binders recognized by T cells and this threshold can be used to select peptides [50 value ranging from 3 to 250 nM. A good epitope should also interact with as many as MHC alleles. Thus, among the total peptides, it was found that only 10 peptides from G protein while 69 peptides from F protein showed interaction with ≥5 MHC Class II alleles. To find out the most probable peptide-based epitopes with better confidence level, selected peptides were further tested using VaxiJen score and peptides having score of ≥0.5 were annotated. Sixteen peptides from F protein and 9 peptides from G protein could be considered as the most potential epitopes for MHC Class II alleles. Among all the most probable epitopes “IPANIGLLGSKISQS” from glycoprotein and “SNIEIGFCLITKRSV” from fusion proteins had the highest VaxiJen scores of 1.4106 and 1.8516, respectively. The most probable epitopic candidates interacting with several MHC class II alleles along with their VaxiJen scores have been shown in While a study reported that a binding affinity  alleles are known and different HLA types are expressed at different frequencies in different ethnicities. Identified epitopes that bind to several MHC alleles would be considered as the best probable epitope only if their combined frequency in a population show good coverage by approaching 100% or close to 100%.http://dx.doi.org/10.1155/2014/402492, represents descending order of percentages of class I coverage by the all populations present in the database.Elicitation of the immune response of the >90%  world population could be covered by the most potential nine epitopes predicted from glycoprotein G. Maximum coverage 98.84%  was founhttp://tools.immuneepitope.org/tools/conservancy/iedb_input) with all G and F protein sequences from different strains of NiV demonstrated that the predicted epitopes were conserved among their respective G and F protein sequences.Conservation analyses of epitopes  were generated Figures  and 5. IThe binding affinities (reflected by the lower binding energies) for the “EWISIVPNFILVRNT” and “GPKVSLIDTSSTITI” to DRB1∗01:01 were estimated to be −6.6 Kcal/mol and −4.6 Kcal/mol, respectively, while for the control peptide “GSDWRFLRGYHQYA”, it was −6.2 Kcal/mol. The binding models generated from Molecular docking using AutoDOCK Vina have been presented in  in silico computational methods after retrieving sequences from the databases. Conserved sequences which were used for the analyses have been presented in The world is now the habitat of more than seven billion people. With the advent of medical technology, new kinds of diseases are also emerging along with new viruses. Developing world, in particular, is more affected by these sorts of diseases. Diseases which have earlier been recognized as zoonotic are now spreading from human to human. However, medical science has always tried to cope with the problems with the pace of replicating disease. Nipah virus infection is one of the reasons of fatalities in humans and livestock in countries like Australia, Bangladesh, India, Malaysia, and Singapore. Nipah outbreaks have resulted in acute respiratory distress syndrome and encephalitis, person-to-person transmission with fatality rates of 40–75% in humans –4. Altho+T lymphocytes (CTL) restrict the spread of infectious agents by recognizing and killing infected cells or secreting specific antiviral cytokines [Most antigens and vaccines trigger not only B cell response but also T cell response. Vaccine induces production of antibodies that are synthesized by B cells and mediates effector functions by binding specifically to a toxin or a pathogen . Howeverytokines . Thus, Tytokines . Peptideytokines , while aytokines , 38 is cytokines . This maytokines . Furtherytokines –42 were Initial observation of the data demonstrated that among all the predicted epitopes “IGFCLITKR,” a F protein derived peptide, has the highest VaxiJen score 2.4587 while “TVNPSLISM” derived from F protein as well as “ISCPNPLPF” and “SLIDTSSTI” from G protein showed interaction with the highest numbers of MHC class I alleles  after ignoring the sequences of glycoprotein G from other isolates from the NiV endemic regions. In the present study, all the available sequences of glycoprotein G and fusion protein F were retrieved from databases and multiple sequence alignment generated conserved sequences which were tested in VaxiJen to predict their probable antigenicity. This, we believe, would generate more acceptable epitope(s) that should be effective universally.However, further investigation of the data revealed that “EWISIVPNF” and “ISIVPNFIL” class I epitopes from F protein have been recognized as the core sequences of four Class II overlapping epitopes that include “SEWISIVPNFILVRN,” “DNSEWISIVPNFILV,” “EWISIVPNFILVRNT,” and “NDNSEWISIVPNFIL” sequences, while epitopes “FILVRNTLI,” “VGILHYEKL,” “IGFCLITKR,” and “YEKLSKIG” were found as the core sequences of “NFILVRNTLISNIEI,” “SVGILHYEKLSKIGL,” “EIGFCLITKRSVICN,” and “SVGILHYEKLSKIGL” epitopes, respectively. On the other hand, in case of G protein, “ITIPANIGL,” “VFYQASFSW,” and “SLIDTSSTI” share sequence with MHC Class II epitopes “ITIPANIGLLGSKIS,” “VFYQASFSWDTMIKF,” and “GPKVSLIDTSSTITI,” while “FPAVGFLVR” and “YFPAVGFLV” as well as “TLYFPAVGF” and “VGFLVRTEF” Class I epitopes share common sequences of Class II epitopes “DTLYFPAVGFLVRTE” and “TLYFPAVGFLVRTEF,” respectively. All these MHC class II epitopes were found to interact with one of the most common HLA alleles, HLA-DRB1∗01:01. These data have been presented in Tables Along with the T-cell epitope, attention was also given to the B-cell epitope, which can induce primary and secondary humoral immunity. Out of the total B cell epitopes (25 from F protein and 18 from G protein), only 7 and 13 epitopes, respectively, from F and G proteins with VaxiJen score of ≥0.5 were selected for further analysis . When T http://www.allelefrequencies.net, the highest frequency was observed in case of HLA-C∗07:02 in Australian, Asian , Europeans, and South Americans compared to the frequencies of other alleles. Thus, for docking, 3D structure of HLA-C∗07:02 was used with the predicted structure of “EWISIVPNF.” As “SLIDTSSTI” epitope did not show any interaction with HLA-C∗07:02 allele, this nonamer was not used for docking. The binding energies of predicted epitope “EWISIVPNF” for its binding to the binding grooves of HLA-C∗07:02 were −6.9 Kcal/mol  showed good MHC class I coverage by the world population even by the population from Southeast Asia where Nipah virus infection has been reported Figures  along wiSupplementary Table 1 (a) represents the coverage of the HLA alleles by different population shown by the epitopes predicted from F protein while (b) indicates overage of the HLA alleles by different population shown by the epitopes predicted from G protein of NiV.


Bioinformatics is now a critical skill in many research and commercial environments as biological data are increasing in both size and complexity. South African researchers recognized this need in the mid-1990s and responded by working with the government as well as international bodies to develop initiatives to build bioinformatics capacity in the country. Significant injections of support from these bodies provided a springboard for the establishment of computational biology units at multiple universities throughout the country, which took on teaching, basic research and support roles. Several challenges were encountered, for example with unreliability of funding, lack of skills, and lack of infrastructure. However, the bioinformatics community worked together to overcome these, and South Africa is now arguably the leading country in bioinformatics on the African continent. Here we discuss how the discipline developed in the country, highlighting the challenges, successes, and lessons learnt. The fields of bioinformatics and computational biology have grown in importance as drivers of research in the life sciences as evidenced by the increasing number of journals and international conferences dedicated to these fields. The need for skills in these areas has grown accordingly in South Africa, as researchers are more frequently awarded grants for projects generating large-scale biological data. Local scientists are generating large and varied datasets including next-generation sequencing , proteomic data, and other data, coupled with rich phenotypic datasets, especially large patient and surveillance cohorts. These research groups seldom have embedded data analysts, so they turn to bioinformatics groups for support. Fortunately, there are well-established bioinformatics and computational biology groups at many of the universities in South Africa, reflecting the significant resources committed to bioinformatics development in the country since the 1990s. In this article, we outline the growth of bioinformatics in South Africa, highlighting the challenges as well as the impact of government support on the development of the field. Where relevant, we also describe lessons learnt.South Africa is a middle-income country , with significant social inequality. While there are resources to develop new scientific fields, demands on government to meet immediate social needs makes neither organic growth nor strategic investment a given. Following the Second World War, state policy and government funding built a strong South African tradition of scientific research. Despite areas of great strength and international leadership, however, there were many neglected areas, policy challenges, and a fragmented science and innovation landscape across both the public and private sector  and "south africa"–"working in partnership with the European Bioinformatics Institute" and “.ac.za”, where “subject area” was [“bioinformatics” or “computational biology”], “chemistry,” “cardiology,” “geology,” “microbiology”. Patents and citations were excluded. We chose to simplify the search and use the broad terms bioinformatics and computational biology, as it is challenging to identify all bioinformatics publications by journal or topic alone. We therefore included the other subjects to demonstrate the relative differences in impact using broad subject terms. It is evident that the number of bioinformatics-related publications in South Africa has increased substantially since 2001, with a sharper rise than other subjects over the same time period.Bioinformatics in South Africa started with a single group at one South African university, but developed rapidly through the injection of funds from the DST via the NBN initiative. The NBN provided a springboard for the development of bioinformatics skills in postgraduate students and bench scientists, and these new initiatives are ensuring a continuation of this momentum. Since then, the Bio-Innovation Unit of the DST has invested in several key bioinformatics-based initiatives, including, the BFG program; the BSP; and the SAHGP. They have contributed to a significant increase in bioinformatics outputs including trained postgraduate students, research publications see , patentsSome of the key achievements of the South African bioinformatics community lie in training, research, and infrastructural capacity development. The universities have collectively trained 130 Masters and 60 PhD students in bioinformatics, many of which have gone on to academic positions or placements in prestigious institutes abroad, including Harvard and the EBI. Bioinformatics courses in South Africa are increasingly being taught by exclusively local trainers, many of whom are frequently invited to give training courses in Africa and abroad. Several groups have contributed to or developed resources used by the international scientific community, one example being the HIV drug resistance database. The strength and international recognition of the community is reflected in the participation of South African academics in key positions in international societies, such as ISCB and ASBCB, partnerships with international research groups in successful funding proposals, and the increase in publications in international bioinformatics journals. One of the highlights, though, is the NIH award for H3ABioNet, which is one of the largest single awards to an African institution.Throughout the history of the development of bioinformatics in SA, many lessons have been learnt, some of which are described above. The take-home message is that commitment to bioinformatics capacity development by the government has been key to the success of many of the bioinformatics units in the country. The step-up provided by the NBN enabled groups in multiple universities to hire dedicated bioinformatics staff members in an environment where universities were not forthcoming in creating new positions. In some cases, the success of the groups convinced the institution of the importance of bioinformatics and the need to support it. The establishment of these bioinformatics units also facilitated the procurement of further funding from the NRF as well as international funding agencies, thus increasing outputs and enabling these units to become internationally competitive in the field.


Purpose. The purpose of this retrospective case series is to evaluate the clinical advantages and limitations of monolithic zirconia restorations for full arch implant supported restorations and report the rate of complications up to 2 years after insertion. Materials and Methods. Fourteen patients received implant placement for monolithic zirconia full arch reconstructions. Four implants were placed in seven arches, eleven arches received six implants, two arches received seven implants, two arches received eight implants, and one arch received nine implants. Results. No implant failures or complications were reported for an implant survival rate of 100% with follow-up ranging from 3 to 24 months. Conclusions. Monolithic zirconia CAD-/CAM-milled framework restorations are a treatment option for full arch restorations over implants, showing a 96% success rate in the present study. Some of the benefits are accuracy, reduced veneering porcelain, and minimal occlusal adjustments. The outcome of the present study showed high success in function, aesthetics, phonetics, and high patient satisfaction. Full arch implant supported restorations have been documented to have high success rates –6. Many Zirconium oxide is a material that has shown increased popularity in contemporary dentistry , 9. ManyTo overcome these problems, CAD/CAM one-block milled monolithic zirconia was introduced as an alternative for the treatment of implant supported full arch reconstructions –5, 12. TShort-term available data indicates that full contour zirconia framework can be used successfully in implant dentistry , 7. SeveThe purpose of this retrospective case series was to evaluate the clinical advantages and limitations of monolithic zirconia restorations for full arch implant supported restorations and report the rate of complications up to 2 years after insertion.Clinical data in this study was obtained from the implant database (ID) in the Ashman Department of Periodontology and Implant Dentistry at New York University College of Dentistry. This dataset was extracted as deidentified information from the routine treatment of patients in the department. The ID was certified by the Office of Quality Assurance at NYUCD. This study is in compliance with the health insurance portability and accountability act (HIPAA).Patients which referred to New York University Ashman Department of Periodontology and Implant Dentistry in need of prosthetic full arch fixed reconstruction in maxilla, mandible, or both were consecutively selected. The inclusion criteria included patients at least 21 years old, with edentulous maxilla and/or mandible and at least four to nine implants needed to be placed and osseointegrated. Fourteen patients  met the inclusion criteria. Each subject selected for this study from the ID had undergone the fabrication of monolithic zirconia frameworks for full arch implant supported reconstructions. Twelve of these patients required maxillary and mandibular full arch reconstruction, and two involved only the maxillary arch. In the two maxillary reconstructions the opposing dentition was in one patient natural teeth and a fixed prosthesis and in the other a complete mandibular denture. A total of twenty-six edentulous arches were restored: fourteen maxillary and twelve mandibular arches. Patients were informed about the prosthetic protocol, risks, and alternatives of treatment.All complications after delivery were recorded at each follow-up visit up to 3 years. Failures were defined as any defect in the restorations that required the fabrication of a new restoration such as fracture and misfitting. Complications were defined as any defect in the restorations that required repair by laboratory technicians or correction of clinicians such as chipping of veneers (lab) and screw loosening (clinician).Diagnostic alginate impressions  were made and poured with model stone . Occlusal rims were fabricated and adjusted intraorally. Interocclusal records, face bow registration, and centric relation records were taken. Casts were articulated and artificial teeth arrangements (ATA) were performed. Additionally, ATA were duplicated to obtain radiographic and surgical guides. Patients were sent for Cone Beam Computed Tomography (CBCT) scans to evaluate bone dimension and implant positioning Figures  and 3.Four to nine dental implants were placed in the edentulous arches using surgical guides. A one-stage surgical procedure was performed according to the implant planning. The surgical protocol followed the manufacturer's instructions. External connection implants were placed in twenty-three arches, and internal connection implants with intermediate abutments were placed in three arches. The healing time prior to the prosthetic phase was 12 weeks. .During the healing period and until the prosthetic phase was completed, patients wore transitional complete dentures.Fixture level impressions were made of polyether impression material  in custom light cure resin trays . The master cast was made of a reproduction of the gingival soft tissue using a polyvinylsiloxane, addition-type silicone  and resin fortified, low expansion die stone . In two patients, in which opposing arches were not restored as full arch reconstructions, alginate impressions were made  with stock disposable perforated trays . Interocclusal registrations were made with wax rims . Face bow registration and centric relation record were taken. Artificial teeth arrangements with acrylic dentures teeth  were placed in an adequate position to achieve esthetics, phonetics, and vertical dimension in occlusion. Bite registration was taken with vinyl polysiloxane material .The laboratory procedures were performed according to the manufacturer's instructions  at an authorized laboratory. The master cast, opposing cast articulated, and artificial teeth arrangement were scanned to determine the interocclusal relationship to the software . According to the corresponding implant type, connectors are milled in titanium to fit in the master cast and scanned again. The prostheses were designed on the software. An epoxy resin prototype was milled and sent for try-in to ensure adequate fit, function, esthetics, and phonetics. After some minor adjustments, the restoration was milled in a monolithic zirconia block. Sixteen of the 26 full arch restorations were digitally cut back on the anterior area to improve esthetics. Characterizations of teeth were made in the monolithic framework . The final restorations were sintered in the oven . Framework fitting was verified in the master cast. Soft tissue ceramic  and ceramic  according to the shade selection was applied on the framework for esthetic results. Additionally, the prosthesis was sintered overnight. Last working step was placement and bonding of the titanium sleeves into the milled zirconia framework. Finally, prostheses were glaze-fired and sent for delivery. Figures , and 9.The final full arch prostheses were clinically verified with one screw test for passive fit. Moreover, periapical radiographs were taken for radiographic examination. All patients approved and agreed with shape and shade of finals restorations. Figures , and 12.Occlusal screws were torqued following manufacturer's instructions. Gutta-percha was placed in all access holes. In screw-retained restorations, a light-cure microhybrid composite  with proper shade was used to close the access hole. In screw-/cement-retained, fixed prostheses were cemented with temporary cement  and excesses were cleaned. The importance of removing the temporary cement and recementing every year, to avoid cement wear and loosening of the prostheses, was explained to the patients.On the day of delivery, alginate impressions were made to fabricate full-cover maxillary night guards. A week later, patients received the night guards and were instructed to wear them at night.Recall appointments were performed after 2 weeks and 3 months after insertion. A yearly appointment is required for clinical and radiographic examination.Fourteen patients received implant placement for monolithic zirconia full arch reconstructions. Four implants were placed in seven arches, eleven arches received six implants, two arches received seven implants, two arches received eight implants, and one arch received nine implants. No implant failures or complications were reported for an implant survival rate of 100% with follow-up ranging from 3 to 24 months .Previous restorations of the total arches were as follows: sixteen had nonrestorable teeth or previously failed fixed prostheses, five arches with teeth and removable partial dentures, three complete dentures, and two metal alloy-acrylic prostheses.Of the twenty-six full arches, twenty-four were implant supported screw-retained, and two full arches were combined implant supported screw-/cement-retained . SeventeImproved clinical performance can be expected to be achieved by using monolithic zirconia restorations , 10, 11.Several different complications have been related to the use of hybrid prostheses with implants, such as fractures of titanium framework and gold alloys over 5 years  and fracHowever, chipping of veneering ceramic is a frequent complication of zirconia-based restorations on teeth and implants , 9, 11 aTranslucency has been considered one of the primary factors in controlling the esthetic outcome of ceramic restorations. Zirconia has been considered an opaque material compared to other dental ceramic. A recent report has shown some degree of translucency in the zirconia, which was less sensitive to thickness compared to lithium disilicate and leucite-free porcelain. However, translucency of the zirconia ceramics also increased exponentially as the thickness decreased . The digWear rates of the enamel opposing zirconia ceramic have been reported, showing cracks or even fractures in all the ridges. The hardness and thickness of enamel, chewing behavior, parafunctional habits, and neuromuscular forces as well as abrasive nature of food can influence clinical wear. Due to the elasticity modulus of 210 GPa of zirconia and hardness of 1200 Vickers Hardness, some enamel wear is expected. Moreover, some reports have shown that polished monolithic zirconia has the lowest wear rate on an enamel antagonist compared to veneered zirconia, glazed zirconia using a glaze spray, monolithic base alloy, or glazed zirconia using glaze ceramic , 19. HowScrew-retained implant restorations are often chosen because they offer better retrievability, decreased space requirements, and healthier soft tissue. On the other hand, cement-retained restorations offer improved occlusal accuracy, enhanced esthetics, increased chances of achieving a passive fit, and decreased instances of retention loss . MoreoveMonolithic zirconia CAD/CAM-milled framework restorations are a treatment option for full arch restorations over implants, showing a 96% success rate in the present study. Some of the benefits are accuracy, reduced veneering porcelain, and minimal occlusal adjustments. The outcome of the present study showed high success in function, aesthetics, phonetics, and high patient satisfaction.A full occlusal contour monolithic framework can diminish chipping of the veneered porcelain. However, the fabrication is technique sensitive and should follow the appropriate steps discussed in this study.The digital cut back for veneer placement in the monolithic zirconia was an effective option to avoid surface roughness that can produce crack propagation and veneer chipping.Twenty-three of 26 restorations were screw-retained due to their retrievability, less biological complications, and easy repair of technical complications. Only, one veneer chipping was found in one these restorations. Of the three cement-retained restorations no complications were reported.Within the limitations of the present study monolithic zirconia CAD/CAM milled prosthetic restorations were a successful treatment option for full arch implants supported restorations.More long-term data studies are required for the full arch monolithic zirconia restorations in order to evaluate success and complications over the time.


Journal of Diabetes Investigation depends on the prompt, careful review of submitted manuscripts. We would like to thank the Editorial Board members, the International Editorial Board members, the Assistant Editorial Board members and the following experts for reviewing manuscripts from October 1, 2013 to August 31, 2014.The publication of invaluable papers in the Hideharu AbeToru AizawaKeiko AraiAtsushi ArakiShin-ichi ArakiYoshimasa AsoAbdul BasitPhillip BrantleyDehong CaiLu CaiJuliana ChanFang-Rong ChangSang Ah ChangTien-Jyun ChangHarn-Shen ChenLiming ChenKen ChiuDong Hyeok ChoJae Hyoung ChoYong Jun ChoiShao-Yuan ChuangDaisuke ChujoSung Wan ChunMakoto DaimonSam J DanielTakahisa DeguchiToshio DoiNabil El-NaggarMasanori EmotoKei FujimotoShimpei FujimotoTomomi FujisawaYukihiro FujitaMichiaki FukuiMitsuo FukushimaKengo FuruichiHiroto FurutaCarlo GalliKathleen M GillespieAtsushi GotoHonglei GuoYusuke HadaAkihiro HamasakiNaotake HashimotoMitsuru HashiramotoTomoshige HayashiYasuaki HayashinoTakumi HirataTakahisa HiroseYushi HirotaTimothy HlaYen-Ping HoYukio HorikawaMasaaki HoshigaMasayuki HosoiChang-Hsun HsiehKuo-Chin HuangCn HuangJee-Fu HuangYanan HuoYou-Cheol HwangChii-Min HwuMd. Auwal IbrahimAtsuhiro IchiharaKazuo InoueKouichi InukaiSan-e IshikawaMasahiro IwasakiMasanori IwaseLijing JiaJyuhn-Huarng JuangTetsuya KakumaNaomi KamimuraHideki KamiyaHideaki KanetoNaoto KatakamiTomonori KatoKoichi KatoTakumi KawaguchiDaiji KawanamiToru KikuchiChul-Hee KimChong Hwa KimSang Soo KimJae Hyeon KimJaetaek KimToshihide KimuraTetsuro KobayashiDaisuke KoyaHidenori KoyamaHiroyuki KubotaAkira KubotaShoen KumeAkio KurodaTakeshi KuroseJohanna KuusistoSatoshi KuwabaraSoo-Heon KwakJoseph KwanHyuk-Sang KwonA LacquanitiHc LamTerence LaoByung-Wan LeeChien-Hsing LeeWon-Young LeeI-Te LeeJenny LeeJi-Hyun LeeYau-Jiunn LeeTing-I LeeHung-Yuan LiQifu LiChou-Ching K LinShaoda LinShu-Fu LinYing LuZhaohui LuShiro MaedaHiroshi MaegawaYuichi MakinoMasafumi MatsudaMunehide MatsuhisaMichihiro MatsumotoSilvia MigliaccioTohru MinaminoJun-ichiro MiyagawaSatoshi MiyataHiroki MizukamiTomoaki MoriokaTatsumi MoriyaSeiichi MunesueShoichiro NagasakaTomoko NakagamiShuhei NakanishiYasuko NakanoTakuma NaritaMasami NemotoDaniel NgMasahiro NishiWataru NishimuraYoshihiko NishioMitsuhiko NodaYoshinori NoguchiMakiko OgataJee-Young OhShinichi OikawaEtsuji OkamotoYoshihisa OkamotoNermin OlgunChristine L OltmanHiroshi OnumaHaruhiko OsawaYen-Chuan OuHarry PowellDavid RodbardYoshifumi SaishoKazuhiko SakaguchiNaoki SakaneHiroshi SakaueKazunori SangoHideyuki SasakiMasataka SataOsamu SekineYi-Jing SheenYunfeng ShenTadao ShibasakiKenichi ShikataHitoshi ShimanoSeiya ShimodaKohji ShiraiHirohito SoneNoriyuki SonodaKazuhiro SugimotoTakashi SugiyamaShoichiro SumiZilin SunYuji TajiriMitsuyoshi TakaharaYoshihiko TakahashiIseki TakamotoFumihiko TakeuchiYasuhiro TakeuchiKohzo TakeyabashiYoshifumi TamuraYasushi TanakaKeiji TanimotoHiroto TerashiYasuo TerauchiHaoming TianKazuyuki TobeSatoru TsujiiDaisuke TsujinoKohjiro UekiHiroyuki UmegakiTatsuhiko UrakamiTakashi UzuJun WadaTakashi WadaHironori WakiChih Yuan WangJong Chul WonJianzhong XiaoKunimasa YagiHiroaki YagyuTomohide YamadaYuichiro YamadaSatoru YamadaSho-ichi YamagishiWari YamamotoRitsuko Yamamoto-HondaShunsuke YamaneTomonori YamanishiShigeo YamashitaTao YangHisafumi YasudaHiroshi YatsuyaNorihide YokoiHirohide YokokawaMasayasu YonedaLiu YongjunNarihito YoshiokaHo Yan Ruby YuPei YuXuefeng YuDalong Zhu


Through constant comparison, the ideas are also compared with requirements of different stakeholders as expressed during a user workshop. This allows for highlighting discussion points for further consideration.The international dialogue on responsible governance of nanotechnologies engages a wide range of actors with conflicting as well as common interests. It is also characterised by a lack of evidence-based data on uncertain risks of in particular engineered nanomaterials. The present paper aims at deepening understanding of the collective decision making context at international level using the grounded theory approach as proposed by Glaser and Strauss in “The Discovery of Grounded Theory” . This st Observation and participation in dialogue on the international governance of nanotechnology and other emerging technologies since the 1990s have inspired a search for concepts to describe the emerging governance structures as well as for setting ethically sound targets for consolidation of these governance structures. Examples of proposed concepts include nanoethics, responsible nanoresearch, nanosafety, precaution, stakeholder dialogue and governance of uncertain risks of nanomaterials. A major criticism of these proposals and initiatives is their arbitrariness: What distinguishes nanotechnology so categorically from other emerging technologies to call for nano-specific ethical principles or governance structures? A decade of research in nanoethics and ethical, legal and societal aspects (ELSA) of nanotechnology and of public and stakeholder dialogue has not resulted in a satisfying answer to this question.What can theories of international relations and deliberative democracy contribute to understanding multistakeholder governance of emerging technologies?What role could software decision support play in democratising such governance?The present article therefore starts from the other end, taking a particular theory of ethically sound global governance as a starting point and then considering its applicability to the case of international governance of emerging technologies including nanotechnology in a secondary instance. The core of the tentative theoretical framework is constituted by Risse’s  review oThe present article aims to contribute to the formulation of a substantive grounded theory [as proposed by Glaser and Strauss ] for theAccording to Glaser and Strauss, the concepts should be analytic—revealing characteristics of the studied entities—and sensitising—yielding meaningful pictures. The formulation of such grounded theory starts with observation and participation in the field of study, in this case the emerging heterogeneous community of stakeholders interested in international governance of nanotechnology. This paper’s authors have been engaged in several research projects on the scientific, ethical, legal and societal (ELSA) and environment, health and safety dimensions of nanotechnology and this experience informs the interpretive lens applied to weave together interdisciplinary theoretical concepts from sociology, ethics and political philosophy with quantitative data on the international governance of nanotechnology from a variety of sources including interviews, workshops, field notes, literature research and surveys. The present article introduces concepts from established theories in international relations and philosophy to analyse data collected from different sources on current practices in international governance of nanotechnology. Glaser and Strauss  promote Comparing incidents applicable to each category;Integrating categories and their properties;Determining the theory, that should be parsimonious in variables and formulation and as wide as possible in scope; andWriting the theoryGlaser and Strauss  explain The analyst can go back and forth between the stages as more data are collected and analysed.Our data sources comprise (a) literature on global governance of nanotechnology . This is certainly responsible for setting the agenda for the debate on risk governance. Already in 2003 it pointed to the potential environmental and safety risks of “Green Goo” and converging technologies and called for a moratorium and a precautionary approach , (scientific) credibility and legitimacy  of the knowledge. This calls for boundary management involving communication, translation and mediation in some form of institution that may utilise a “boundary object” such as co-development of a model or scenario by representatives of all three groups. The review reveals some unresolved issues:These are key aspects in the case of international norm creation for nanomaterials. Despite several years of stakeholder and policy dialogue, it is not always clear which decision maker needs what kind of data. Governance initiatives are taken at national, transnational (EU) and global level, but not very well coordinated, according to some interviewed industrialists and regulators . The tool performs a semi-quantitative benefit-risk evaluation of nano-enabled products over their life-cycle and has low data requirements. Specifically, the model looks at environmental, social and economic benefits vs. ecological, occupational and consumer risks. Tier 2 of SUNDS includes the same components as Tier 1, but manifests a higher level of complexity and certainly has significantly higher data requirements. It comprises two modules: (a) risk control (RC) module, and (b) socioeconomic assessment (SEA) module. The RC module integrates quantitative ecological and human health risk assessment tools and facilitates users to select technological alternatives and risk management measures to reduce risks based on efficiency of mitigation, technological maturity and cost. The SEA module will compare the monetised benefits and costs of nano-enabled products with conventional alternatives. The SUNDS tool aims to evaluate the benefits and the risks of nano-enabled products along their life cycle. International governance of nanotechnology encompasses higher levels of social organisation and different governance concerns that can be addressed by further expanding the SUNDS tool.As part of the stakeholder needs solicitation for the abovementioned tool, a roundtable was organised on “Computer guidance and decision making for safe and sustainable nanomaterials”.The interviews on stakeholder needs for the abovementioned tool design also addressed a similar potential role in international governance: “According to some interviewed policy makers, they are not likely to use software decision support tools in preparing political decisions on regulating nanomaterials, especially not in the short term. According to other stakeholders, policy makers could benefit from tools that indicate whether they should take action on adapting regulation, or that support international harmonisation of nanomaterials regulations”  or association in data collection, cooperation in subsidised research consortiaReduction in insurance premiums—insurers would be interested in a system that reflect the emergence of risks that would call for increases in insurance premiumsPublic image of the (large) company (sustainability)Communication with clients and partners in value chain (SMEs)Improving its own productTraining by researchers and consultants how to use the toolsIntermediaries, associations and consortia or networks are commonly used to generate data for norm creation in a cooperative effort. Incentives may be:Industrial users may require protection of proprietary data costing a lot of moneyIndustry wants to avoid giving away expensive data to competitors—need accommodation for sharing thisNobody wants to take the lead in opening up the informationDesign and data requirements of SMEs and researchers clashIndustrial users may lack the skills to use the toolsBarriers to participation of MNCs in the epistemic community include:MNC overlap with epistemic communities, because they employ risk assessment specialist carrying out tests of nanomaterials in the company’s R&D and manufacturing. They are encouraged by policy makers to share these test results with the international community to facilitate the legislative process. A bottleneck hampering such contributions is the need for protection of proprietary data and to ensure that the costs are shared by all competitors and no one is given a free ride. Is it possible to address this issue specific to public–private partnership during the norm creation phase by smart design of the wiki-tool or is a formal legislative framework a prerequisite?A variety of companies participates or contributes to the value chain from nanomaterials R&D and production all the way to the waste processing stage of products incorporating nanomaterials. This includes large  companies as well as SMEs, service providers such as instrument manufacturers, banks and insurance companies and industrial associations. As discussed above some of those companies actively engage in dialogue on international norm creation, either directly (mostly associations and large companies) or indirectly through participating in working groups organised by associations , and fixing these weights in the system. Another way would be to include these average weights as default values in the tool, but allowing users to insert their own weights and compare those with the average of their own and other stakeholder groups. The second option would enhance transparency of how different values and preferences may influence international norm creation on nanomaterials.As argued in the section on nanotech sustainability from insurability, insurance companies play the role of proxy regulators during the international norm creation phase. The discussions during the workshop suggest that they need data to support insurance decisions (including estimates of uncertainty) such as occupational health and safety and catastrophic events . They currently seldom make use of decision support tools or computer models in supporting their decision making on insurance policies. One insurance company represented in the user workshop has used PraediCAT,On the regulatory side, national, transnational (e.g. EU) and international policy makers (e.g. OECD) participate in international norm creation, including representatives of states as well as the European Parliament. Authorities participate in norm implementation. During norm creation, regulators are interested in a category approach enabling read across to drive risk assessment outcomes for risk prioritisation and identification of testing needs. They would be interested in a system that reflect the emergence of risks that would call for new regulation. , where research groups trained civil society organisations enabling them to formulate positions and participate in the dialogue on governance of nanomaterials and nanotechnologies.20The stakeholder needs elicitation in this SUN project has not covered advocacy networks. An interviewed regulator is interested in a tool “that can make it easier for NGOs and watchdogs to monitor compliance of companies to the norms during (the future phase of) implementation.”Other studies may suggest how a wiki-type software decision support tool could enable advocacy networks to contribute to the current norm creation and whether there would be any interest in such contributions from their side. For example, Invernizzi  cites a Returning to Habermas’ vision on a dual national and global citizenship for all, calls for discussion of another type of actor that has so far been excluded from the analysis in this article: the elected representatives of citizens. Some initiatives and dialogue projects on nanotechnology have attempted to foster direct democracy  sustainable manufacturing to select less hazardous materials early in R&D. Some large companies consider this in their own self-interest, while most are not intrinsically motivated for this. Intermediaries and insurance companies are already collecting data from their members that could be inserted in the tool and contribute to supporting the creation of international evidence based norms. They may offer support and benefits to the companies. Industry could also be pressed into using non-regulatory risk assessment through threats of litigation on manufacturing as developed in the USA. Suppliers and partners in the value chain as well as industrial associations and consortia can offer support or a platform for generating common data sets.Most SMEs (and large industry) will not use a tool unless it is compulsory, limiting its use to the norm implementation phase. Regulatory compliance is very important for them. The tool could be used to demonstrate compliance with existing legislation including REACH, cosmetics and biocides regulations in Europe.The discussions during the workshop suggest that in the long term, a decision support system may contribute to epistemic coordination through uncertainty reduction Haas  by graduMixing values with data is problematic because of legal and political reasons. It is important to clearly separate them in the tool. Examples of values include the following: The definition of nanomaterial imposed by regulators determines the scope of what must be tested. This is a political more than a science-based decision. Data quality can be determined through protocols imposed by regulators and scientific expert judgement . The precautionary principle or approach is a core but controversial concept in governance of nanomaterials, that should not be incorporated in the tool design. Instead, uncertainties should be made explicit allowing users to be as precautionary as they wish. The level of acceptable risk for the different endpoints can either be based on current regulator preferences or on scientific expert community judgement. The tool designers can insert a fixed set of endpoints based on regulation and let the scientific community define additional endpoints. Which weights will be included for the different endpoints as default and which options the user will have is an open question.Recall the first central research question: “What can theories of international relations and deliberative democracy contribute to understanding multistakeholder governance of emerging technologies?”Risse , Haas 1 and otheIn response to the second central research question: “What role could software decision support play in democratising such governance?”, there appears to be room for wiki-like software decision support tools in the international governance of nanomaterials. This three-stage process consists of past agenda setting, current norm creation and current as well as future norm implementation. A well-designed wiki-tool could bring together test data and values from all experts and stakeholders and increase the transparency of the scientific data and values taken into account in international norm creation. Now and in the future, the tool could be used by all to demonstrate, verify and monitor compliance with the norms.Comparing the sketched international governance context and the wiki-option with the current design of the SUNDS tool highlights the following discussion points for further consideration:How can citizens be empowered in international governance of nanomaterials  through decision support?What would be the optimal democratic and technically feasible option to ensure good data quality and the protection of proprietary data while allowing anyone to toggle with the weights?Is it possible to allow users to insert their own weights and compare them with the average of their own peer group and other stakeholders?Could the design be flexible enough to accommodate different emerging legislative frameworks ?