Amphimedon queenslandica. Auto- and allografts were established, observed and sampled over a period of three days, over which time the grafts either rejected or accepted, depending on the identity of the paired individuals, in a replicable and predictable manner. Fourteen transcriptomes were generated that spanned the auto- and allograft responses. Self grafts fuse completely in under three days, and the process appears to be controlled by relatively few genes. In contrast, nonself grafting results in a complete lack of fusion after three days, and appears to involve a broad downregulation of normal biological processes, rather than the mounting of an intense defensive response.Sponges, despite their simple body plan, discriminate between self and nonself with remarkable specificity. Sponge grafting experiments simulate the effects of natural self or nonself contact under laboratory conditions. Here we take a transcriptomic approach to investigate the temporal response to self and nonself grafts in the marine demosponge  Hydractinia echinata must be shared between two or more colonies [Clathria prolifera were identified at relatively high (20%) frequencies within the population [Marine benthic ecosystems often are densely-populated and have remarkable levels of animal biodiversity . In such a crowded environment, space can become a limiting resource, and sessile invertebrates in particular often face intense competition for habitat and growth space. For example, one study determined that 42% of microhabitats  for the colonial hydrozoan colonies . Similarpulation ,4,5. Howpulation . For thipulation . The decThe sponge has been a useful model animal for the study of cell adhesion and self-nonself recognition for almost 150 years, with grafting experiments first described in 1869 ,17,18,19Clathria prolifera, which responds to allografting in two to six hours [Callyspongia diffusa, which can take up to a week to react [Typical self grafts that undergo fusion are characterised by the breakdown of the pinacoderm layers separating the two pieces of sponge, with the interface between the graft donors becoming invisible over time ,15,21. Rix hours , or compto react ,24,25,26to react ,20,24,25to react ,27,28,29to react ,23,29,30to react ,24,25,26to react , betweento react ,25,28. Tto react .Amphimedon queenslandica. We established and observed grafts over a three-day period, over which time each graft underwent either a fusion or a rejection reaction. We periodically took cellular samples from the graft interface. Transcriptomes generated from these samples were analysed to identify the broader functional changes that distinguish the self grafting response from the nonself grafting response in A. queenslandica. To our knowledge, this represents the first longitudinal, high-throughput sequencing approach applied to understanding the molecular allorecognition response in sponges.Here we analyse the expression dynamics of the auto- and allograft response in the demosponge A. queenslandica individuals, and analysed the qualitative and quantitative changes in expression that occurred across the graft time course.Sponge grafting experiments have been well-described in the literature since 1869 [A. queenslandica. Samples were collected from the graft interface at each time point.Grafts were established between four pairs of sponge individuals, with each pairing producing one nonself and two self  time courses. Multiple grafts were created to allow separate analysis at each time point to avoid disturbance during observation. The grafts were observed at 12, 24, 48 and 72 h post grafting (hpg) to determine the nature and timing of the physiological response to self or nonself contact in For six of the eight autograft time courses, early signs of fusion were first observed at 12 hpg. Grafts from the seventh sponge initiated fusion by 24 hpg, and the eighth by 48 hpg . Bonds bTwelve hours after grafting, all four allograft samples remained unfused. However, at 24 hpg several of the samples exhibited signs of partial attachment . Here, wA. queenslandica genes  [One of the four graft experiments was selected for whole-transcriptome RNA sequencing (RNA-Seq) and analysis. It comprised of two self and one nonself time courses, each sampled at 0, 12, 24, 48 and 72 hpg. Samples are named here by time point  and graft type . Final sequencing datasets each contained between 17.5 (T24AB_C) and 27.8 (Donor A) million reads . The aveBioMart) . Read trGenetic identity, rather than immune state, appears to be the primary factor promoting gene expression differences between samples, when considering the most dynamically-expressed genes across all samples. In a principal component analysis (PCA) , the AA A. queenslandica genome to determine the read counts per Aqu2.1 gene model. These counts were then used to identify genes exhibiting statistically significant fold changes between successive pairs of time points. The two self time courses, AA and BB, were analysed separately in light of the finding that between-individual differences were the primary source of variance between samples , and our morphological results are consistent with past findings. Of the eight examined self graft time courses, fusion was observed for all samples by 48 hpg. Observed variability in the onset time of initial fusion likely represents inter-individual variation, but possibly also variability in graft contact surfaces and/or failure to observe early weak bonds between grafted pieces. In all four nonself graft time courses, rejection had occurred by 72 hpg. Sponge pieces in the rejected grafts remained alive and healthy, with no signs of necrosis obvious to the naked eye. Three of the four nonself graft time courses exhibited signs of minor attachment between 12 and 24 hpg. Here, weak bonds appeared to join the two pieces of sponge, and light force was required to separate the two pieces. The bonds between the pieces may not represent true early fusion, but rather, for example, fibrous material produced during graft rejection that randomly interlaced due to proximity of the two sponge pieces. However, it may be that a degree of fusion is required early in the rejection process, to allow cellular infiltration of the graft interface, direct cell-cell contact between cells of the opposing individuals, and subsequent rejection. Such a phenomenon has been reported elsewhere, with tissue-like bridges spanning the nonself graft interface in other sponge species ,34,35. B diffusa , further diffusa ). Early le pairs . A similPCA of the most dynamically expressed genes across the three graft time courses revealed greater divergence between sponge individuals than between graft conditions  . This beRelatively low numbers of genes were found to be differentially expressed between successive autograft time points. The allograft time course exhibited more dynamic expression across time, particularly between the 24 and 48 hpg time points . It is nSixty-five percent of differentially expressed genes identified within the nonself time course were found to be downregulated at the point of detection. These downregulated genes were statistically enriched for GO terms associated with key biological processes such as cell signalling, transcription and translation, protein and molecular transport, and other metabolic processes . This maBotryllus schlosseri. In this species, rejection reactions are asymmetric, where one graft partner develops morphological signs of rejection , while the other partner does not  [B. schlosseri graft showed limited gene upregulation relative to the naive state, but extensive downregulation of genes involved in protein biosynthesis, cell structure and motility, and immune function; rejecting individuals showed limited changes relative to the naive state [A. queenslandica. Additionally, although no obvious physiological signs of a ‘rejected/rejecting’ hierarchy have been noted within A. queenslandica, it is possible that some molecular, genetic or physiological hierarchy may indeed exist.A transcriptional shutdown in response to graft rejection has also been reported in microarray analyses of gene expression in the ascidian ividual) . Rejecteve state . Here itve state . It is pA. queenslandica adults (‘donors’) from Shark Bay, Heron Island Reef  [Four grafting experiments were performed in total. Four pairs of non-adjacent stralia)  were remFor one graft experiment, samples were taken from the graft interface and preserved in RNA Later (Ambion) for whole-transcriptome sequencing, both from the two donor sponges prior to graft establishment and from the auto- and allografts at each time point. A total of 200 mg sponge was used per RNA extraction . RNA was extracted using Tri Reagent  following manufacturer’s guidelines. RNA quantity and quality was checked using a Qubit 2.0  and Bioanalyser 2100 .RNA was sequenced by Macrogen Inc.  using a polyA-selection, 100 base pair (bp), paired-end, unstranded Illumina HiSeq 2000 protocol. Eighteen libraries were multiplexed into a single Illumina flow cell lane. For the T24AB time point, three different RNA samples were sequenced from the original sponge sample, due to initial concerns about RNA quality and quantity. However, all resulting T24AB samples were deemed to be of sufficient quality for this analysis and their reads were pooled for subsequent analyses. Library quality was assessed using FastQC 0.10.0  , and pooA. queenslandica Aqu2.1 gene models [Trimmed reads were mapped to the e models  using the models , and thep ≤ 0.01) changes of four-fold or greater (log2) expression were identified using EdgeR’s GLM functionality.Independent filtering was performed to remove statistically uninformative genes and improve multiple testing correction outcomes , using tA. queenslandica genome. Enriched GO terms were clustered based on semantic similarity (SimRel measure) using REVIGO [GO annotation of the Aqu2.1 gene models was performed using Blast2GO version 2.8  and annog REVIGO . Similar2 fold changes between genes of interest were generated using the R function heatmap.2 within the gplots package [Venn diagrams were generated using the online tool Venny . Unscale package  using deA. queenslandica appears to occur over three days. Self grafts initiated fusion between 12 to 48 hpg, and the graft interface had nearly completely disappeared by 72 hpg. The nonself graft response does not appear to be aggressive . Instead, both partners remain alive and healthy for the duration of the graft response. Allografts may undergo a period of transitory fusion between 12 and 48 hpg, where weak bonds formed between the sponge pieces, possibly to allow direct cell-cell contact between the rejecting sponges. Analysis of the global transcriptional changes occurring during this time suggests that there is not a large degree of defensive gene activation in the vicinity of the allograft, instead normal cell physiological processes, communication and proliferation appear to be repressed.The process of self-nonself grafting in


The effect of intravitreal bevacizumab in a rare case of retinal dystrophy with secondary cystoid macular edema"", authors Macovei Mioara-Laura, Nica Maria-Alexandra, was published in Romanian Journal of Ophthalmology, 2017; 61(2): 123-127. The authors would like to change the text “oral acetazolamide 500 mg daily and aspacardin 100 mg daily” into “Acetazolamida Arena oral 250mg x 2/ daily, Aspacardin Terapia oral 39mg/ 12mg x2/ daily”.The article The above-mentioned change should be made at all the levels, starting with the journal’s web page – www.rjo.ro and ending with the database the journal is indexed in (PubMed). The publishers regret any inconvenience caused by this change.


Elysia timida and its sole food source, Acetabularia acetabulum were examined to determine whether or not starch, a combination of amylose and amylopectin and the main photosynthate produced by A. acetabulum, is produced by the stolen plastids and whether it accumulates within individual kleptoplasts, providing an energy larder, made available to the slug at a later time.Solar-powered sea slugs are famed for their ability to survive starvation due to incorporated algal chloroplasts. It is well established that algal-derived carbon can be traced in numerous slug-derived compounds, showing that slugs utilize the photosynthates produced by incorporated plastids. Recently, a new hypothesis suggests that the photosynthates produced are not continuously made available to the slug. Instead, at least some of the plastid’s photosynthetic products are stored in the plastid itself and only later become available to the slug. The long-term plastid-retaining slug, Elysia timida throughout a starvation period were stained with Lugol’s Iodine solution, a well-known stain for starch granules in plants. We present here for the first time, an increase in amylose concentration, within the slug’s digestive gland cells during a starvation period, followed by a sharp decrease. Chemically blocking photosynthesis in these tissues resulted in no observable starch, indicating that the starch in untreated animals is a product of photosynthetic activity.Histological sections of This suggests that kleptoplasts function as both, a nutritive producer and storage device, holding onto the polysaccharides they produce for a certain time until they are finally available and used by the starving slug to withstand extended starvation periods. Sacoglossan sea slugs (Heterobranchia: Gastropoda) are also known as “solar-powered sea slugs” and “leaves that crawl” due to some members’ ability to steal chloroplasts (kleptoplasts) from their algal food and retain them for many months –5. As th14C or 13C were conducted [14C was lower in specimens kept in the dark, which presumably has no effect on heterotrophically derived 14C, the amount of photosynthetically derived 14C was determined, demonstrating the metabolic movement of photosynthetically derived carbon into multiple slug-produced compounds , although their longevity may be compromised [Chlorophytes, the algae most often ingested by sacoglossans, produce numerous photosynthates including carbohydrates, lipids and proteins , 10–13. onducted , 14–19, onducted , pigmentonducted , mucus [onducted , amino aonducted ). Since mpounds  and Fetovaia, Elba, Italy (April 2015) at one to two meters depth. Specimens were kept in groups of 25, in aerated tanks containing roughly 3 L freshly prepared artificial seawater (changed every other day). Artificial lighting for 12 h each day provided full-range lighting measuring about 220 μE−2s−1 at the water’s surface to mimic natural conditions and the optimal irradiance [A. acetabulum for two weeks to acclimate to laboratory conditions and to assure feeding.radiance , 37. BotPrior to fixation, the maximum quantum yield (PAM or Fv/Fm) was measured three times with a Pulse Amplitude Modulated fluorometer (Diving PAM) for each animal in order to compute a mean value . This method assesses photosynthetic efficiency in living tissues. PAM values were also measured for specimens treated with the photosynthetic inhibitor Monolinuron – diluted to 2 μg.ml-1 with filtered seawater, in order to determine if photosynthesis was still occurring in the slugs and how it decreased with the continuous presence of the blocker. The monolinuron and seawater mixture was prepared freshly and changed with the water every other day to ensure continuous exposure.A. acetabulum.Forty individuals were starved for each time series: the fall population, spring population and monolinuron. Two specimens  or three specimens (spring population) were first photographed Fig.  and thenTen regions were chosen throughout the slug, in order to analyze different areas containing digestive gland tubules. For each region, two pictures were taken and analyzed in order to estimate the mean relative starch percentage value (RSP). This percentage value was computed using Adobe Photoshop (version CS5.1) by first deleting all non-digestive gland tubule tissue and recording the total digestive gland tubule area Fig. . Then eaE. timida specimens from Elba  did not survive more than 30 days of starvation (except for some specimens that managed 42 days)  Figs. .Fig. 4HiIn the case of the Blanes fall population, the relative starch percentage (RSP) increases until 42 days Fig. , reachinThe Elba spring population also showed an increase in the RSP until 15 days of starvation Fig. . AlthougMonolinuron-treated samples did not contain amylose, at any time point during the starvation period  Figs.  and 4b. E. timida adults, showing that this photosynthate is produced and maintained by A. acetabulum kleptoplasts. Unstarved (control) slugs contained only small amounts of starch. After 3 days (in the case of the Elba spring population) or 10 to 21 days , incorporated chloroplasts had the time to produce photosynthates, which accumulated as starch within the plastid  was found amongst A. acetabulum that were young and completely uncalcified whereas the spring population (Elba) was collected from algae that had already grown long stalks and were already forming caps [A. acetabulum during April, on the island of Elba, Italy. Calcified A. acetabulum stalks present a physical barrier that prevents E. timida from feeding [Acetabularia lifecycle and is followed by a planktonic stage that presumably cannot be eaten by E. timida. Calcified stalks and cap-formation indicate a lower amount of food meaning slugs take in fewer kleptoplasts within the spring population (Elba) slugs, which may also account for a faster decrease in the spring RSP. Kleptoplast abundance has not been described from adult specimens collected in different seasons, so this hypothesis warrants further investigation.Another effect due to the sampling period is that ing caps . This aling caps , who des feeding . Cap-forThe immediate decrease in amylose observed in monolinuron-treated specimens shows that starch production is the result of the ongoing photosynthesis in incorporated chloroplasts. Furthermore, the slugs are not able to face longer starvation periods and show drastically reduced longevity, dying after 21 days. This contradicts previous findings , 24,whicA. acetabulum plastids and starch production in E. timida. The exact effects of monolinuron in these tissues is still unknown, and it is possible that these plastids may still be producing simple sugars such as fructose and glucose, the prerequisite components of amylose and amylopectin, but not producing starch itself. If starch was produced in some of the unblocked plastids, as Christa et al. [Christa et al.  state tha et al.  suggest,E. timida specimens were able to incorporate photosynthetically active chloroplasts, which increase their lifespan during starvation via starch production, we conclude that these incorporated chloroplasts provide nutrition and appear essential for slugs in longer starvation periods. The slug as a direct recipient of these photosynthates is a more likely scenario, since the isotopic radio labeling studies showed 14C labeled carbon in numerous slug produced compounds [By demonstrating that ompounds , 14–18, This study is the first to examine starch production by the chloroplasts stolen and stored in long-term retaining sacoglossan slugs. Previous findings have uncovered algal-derived carbon in slug tissues unrelated to digestion, however it was unknown when and in which form fixed carbon was made available to the starving slug. We now know that not all of the photosynthates produced are immediately transported to slug tissue, rather some accumulate in the kleptoplasts as starch grains. These starch grains increase in size until about halfway through the maximum starvation time experienced by the slugs and subsequently decrease rapidly. This suggests that starving slugs receive the nutritive benefits of the enslaved plastids after an extended time period and that these chloroplasts do indeed help the slug withstand long starvation periods. By using the photosynthetic blocker monolinuron and finding the chloroplasts lack starch, we confirm here that the starch buildup is in fact due to photosynthesis occurring in these chloroplasts. While many aspects of this enigmatic interaction between slugs and kleptoplasts remain unknown, this study directly indicates a major benefit the slug receives from its sequestered chloroplasts. This directly supports the causative hypothesis that some species of sacoglossan slugs can withstand extended starvation due to their sequestered plastids.


Food & Nutrition Research (FNR), I wish to recognize the energy and passion of everyone involved in contributing to and operating this journal. Human capital is responsible for the creation of FNR as the dynamic publication of it spans from the authors who contribute their work to our editors to our reviewers – all of whom seek to work in a timely manner, while facing their own demanding time schedules. FNR is certainly ahead of the times, delivering up-to-date nutrition research that now reaches nearly the entire globe online. There are many great and powerful moments that go on behind the scenes of FNR and I have had the opportunity to both experience these moments and celebrate them over the last 2 years.Having completed my second year as Editor-in-Chief of As 2016 comes to an end, we can conclude that it marks a successful and busy year. In terms of output, FNR has been productive, 100 papers published for the first time, which represents a significant growth in comparison with 2015.In other areas of growth and change, our publisher Co-Action Publishing joined the Taylor & Francis Group, one of the world's leading publishing houses. I believe that this new publishing environment along with the continued support of the Swedish Nutrition Foundation will contribute actively to the further development and direction of FNR. As part of the many steps being taken to manage the growth of the journal, new editors are being appointed and the FNR editorial board is being updated. Editors and the board are expected to contribute actively to developing the journal and to its governance. We will be holding a virtual meeting in the coming months to disseminate all our plans and receive feedback from the board members.On a personal level, I would like to acknowledge the considerable contribution and efforts of the editorial team, namely the editors Per Ole Iversen, Wendy Russell, and Kenneth Maleta and our newly appointed Statistical Advisor Abhik Ghosh. Moreover, I wish to express my gratitude to Veronica Svärd and Caroline Sutton of Co-Action Publishing, and Anneli Hovstadius and Nina Jansson from the Swedish Nutrition Foundation, for their considerable support and patience as we have grown the journal together. Looking forward, the focus of FNR will remain unchanged, namely on publishing Nutrition Research as outlined on the FNR website.In conclusion, we thank all the authors and reviewers for their efforts in developing these studies to a satisfactory outcome. We invite nutrition researchers to submit their original work to FNR and to future special issues. Please do not hesitate to contact me or other members of the editorial team if you feel you can contribute to the further development of the journal.Best wishes for a wonderful holiday season and have a very happy new year 2017!Asim K. DuttaroyEditor-in-Chief


Zika virus (ZIKV) is a mosquito-borne flavivirus distributed all over Africa, South America and Asia. The infection with the virus may cause acute febrile sickness that clinically resembles dengue fever, yet there is no vaccine, no satisfactory treatment, and no means of evaluating the risk of the disease or prognosis in the infected people. In the present study, the efficacy of the host's immune response in reducing the risk of infectious diseases was taken into account to carry out immuno-informatics driven epitope screening strategy of vaccine candidates against ZIKV. In this study, HLA distribution analysis was done to ensure the coverage of the vast majority of the population. Systematic screening of effective dominant immunogens was done with the help of Immune Epitope & ABCPred databases. The outcomes suggested that the predicted epitopes may be protective immunogens with highly conserved sequences and bear potential to induce both protective neutralizing antibodies, T & B cell responses. A total of 25 CD4+ and 16 CD8+ peptides were screened for T-cell mediated immunity. The predicted epitope "TGLDFSDLYYLTMNNKHWLV" was selected as a highly immunogenic epitope for humoral immunity. These peptides were further screened as non-toxic, immunogenic and non-mutated residues of envelop viral protein. The predicted epitope could work as suitable candidate(s) for peptide based vaccine development. Further, experimental validation of these epitopes is warranted to ensure the potential of B- and T-cells stimulation for their efficient use as vaccine candidates, and as diagnostic agents against ZIKV. ZIKV: Zika virus; NCR: non-coding regions; ORF: open reading frame; MHC: major histocompatibility complex; HLA: human leukocyte antigenZika virus (ZIKV), a pathogenic vector borne flavivirus is a member of the Flaviviridae family, transmitted by the Aedes mosquitoes and is known to cause Zika fever. This virus was first identified in the blood of a sentinel Rhesus monkey during a study on Yellow fever in the Zika forest of Uganda in 1947. This was followed by subsequent isolation of this virus from a pool of Aedes mosquitoes of African region . Pregnain silico approach. This might be helpful in developing peptide based vaccines against the Zika virus.Due to lack of rapid diagnostic techniques, effective treatment and availability of novel vaccines against the Zika virus has been a public health emergency of the international concern declared by the World Health Organization . Hence,www.uniprot.org). The retrieved sequence (A0A060H177_9FLAV E protein) was a further subjected to immunogenicity assessment and epitope prediction. The 504 length amino acid sequence of viral envelope (E) protein (GenBank: AIC06934.1 & RCSB PDB ID: 5IZ7), involved in the host cell binding and fusion activity was retrieved from UniprotKB Database (http://www.ddg-pharmfac.net/vaxijen/VaxiJen/VaxiJen.html) was used for the evaluation of immunogenicity of the retrieved protein sequence. This server works on Auto Cross Covariance (ACC) algorithm that predicts protective antigens, tumor antigens, and subunit vaccines with the precision level of 70 to 89 % for the discrimination between antigens and non-antigens .www.imtech.res.in/abcpred) by selecting the window length of 20 amino acids, which were recommended for humaoral immunity. This server is based on information - processing algorithms inspired by the biological nervous system. The server ranks the epitopes as per their respective scores. The higher score of the peptides means a greater probability of becoming most suitable immunogenic epitope .www.iedb.org)  tool IEDB online server . Human http://www.ddg-pharmfac.net/vaxijen/VaxiJen/VaxiJen.html)The immunogenicity of both B-cell and T-cell predicted epitopes was evaluated using a VexiJen V2.0 online server  server was used to determine the best solvent for the predicted peptide based on its amino acid sequence.The Peptide Property Calculator (http://tools.immuneepitope.org/tools/population/iedb_input) of T-cell response, the peptides with wide range of HLA binding specificities result in wider population coverage in terms of geographical expansion. The population coverage rate of the predicted epitopes was calculated by employing the IEDB population coverage tool . The prThe immunogenicity of the viral envelope protein was ensured using VaxiJen V2.0 online server, keeping the threshold at 0.4. The results obtained suggest that the viral sequence is a probable antigen with a score of 0.6178. The prediction and identification of B-cell epitopes in target antigens, being a key step in the epitope based vaccine development, was performed by using ABCPred server. Based on the artificial neural network (ANN) method, the epitopes predicted by ABCPred were in the descending order of their scores, depicting that the top most has the best binding affinity. Therefore, the first (top) epitope 'TGLDFSDLYYLTMNNKHWLV' with the score 0.90 at the position 194 to 213 showing the highest probability to be an antigenic determinant, was selected for further study. MHC-II restricted CD4+ T-cells activation plays an important role in initiating and upholding a proficient antibody response or cytotoxic T-lymphocytes (CTL) response. Further, MHC-I restricted CD8+ CTLs have a vital role in combating the viral infection. Thus, the comprehending principles of T-cell activation and epitope-based vaccine design, direct the identification of helper T-cell epitopes and CTL epitopes. The helper T-cell epitopes and CTL epitopes in the 'Env' protein of Zika virus were identified using the IEDB online tool. The immunogenic peptides were ranked in accordance with their consensus percentile rank from SWISS-PROT program. The lowest percentile rank demonstrated as the best binders. By selecting the first (lowest percentile rank) epitope for each of HLA class-II and HLA class-I gave 63 helper T-cell epitopes (Supplementary Information: Table S1) and 76 CTL epitopes (Supplementary Information: Table S2).http://www.ddg-pharmfac.net/vaxijen/VaxiJen/VaxiJen.html). This tool predicts the antigenicity of a peptide on the basis of amino acid properties and their positions in the peptide. The results demonstrated that the identified B-cell epitope is immunogenic in nature with score of 1.2758, and among all the identified T-cell epitopes, a total of 16 HLA class-II and 25 HLA class-I epitopes were found to be conserved as well as immunogenic (Supplementary Information: Table S3). Peptides with strong antigenicity are always better candidates to be B- and T-cell epitopes than those with weak antigenicity. Hence, the assessment of immunogenicity of the identified peptides was performed using the VaxiJen V2.0 server  was further tested for the toxicity analysis and medium solvency. All the predicted immunogenic epitopes were non-toxic and were classified as acidic, basic or neutral Table 1. Acidic HLA allocation varies among diverse ethnic groups spread over different geographical regions of the world. Therefore, while designing a potent vaccine, population coverage must be taken into account to cover maximum possible populations. In this study, all alleles in IEDB database were identified as best binders with the predicted epitopes and were used to establish the population coverage for these epitopes Table 2. A signiin silico strategy might prove as a boon for the experimental biologists in rapid screening and identification of probable vaccine candidates. T-lymphocytes play a central role in the generation of a protective immune response in many microbial infections  instead of whole protein as a vaccine has been confirmed to represent the complete antigenicity of any protein . Also, al., 2003), hence Recently, a similar approach was used for the identification of highly immunogenic peptides of Env protein of Zika virus  and sugThe sequence of the viral envelope protein was used for the evaluation of the viral immunogenicity to predict the vaccine candidates against the immune-responses of B- and T-cells. The B-cell epitope and the best/highest percentile score generated a single epitope 'TGLDFSDLYYLTMNNKHWLV' at a position of 194 to 213 with immunogenic score 1.2758. The putative CD4+ & CD8+ T-cell core epitopic sequences for all the HLA super types were predicted. We also checked our predicted immunogenic epitopes for the conserved regions and further subjected them to ToxinPred server for the differentiation of toxic or non-toxic peptides. The assessment of the immunogenicity and toxicity properties of the putative epitopes using various online servers showed that the predicted epitopes were immunogenic in nature, and no sequence variability was detected in the region containing the potential epitopes.The epitope NTKNGSISLMCLALG at position 478-492 for HLA Class-II and the epitope RLSSGHLKCR at position 283-292 for HLA Class-I was found to have the highest antigenic score of 1.9904 and 2.0901, respectively, and both were found to be basic in nature Table 1. The putative epitopes have excellent worldwide population coverage as depicted in Table 2in silico approaches by immunologists has helped to speed up the in vitro, in vivo and ex vivo studies by cutting down the large epitope dataset into a smaller one that needs to be confirmed by the experimental analysis. The predictions made by the present study on the primary amino acid sequence of Zika virus should help other researchers to experimentally verify the immuno-dominant epitopes fundamental to the understanding of the pathobiology of Zika virus leading to effective vaccine development.The application of in silico strategy might prove as a boon for the experimental biologists in rapid screening and identification of probable vaccine candidates. The proposed non-toxic epitopes would be a relevant representative of a large proportion of the human population. This work indicates that the proposed epitopes prompt the future vaccine development against the Zika virus. However, the potential of the screened epitopes by experimental validation is warranted.The data from the current work on B- and T-cell peptide epitopes for 'Env' protein of Zika virus might prove a milestone in the development of vaccines to combat Zika virus induced infection. The suggested Essam Mohammed Janahia and Shafiul Haque  contributed equally as corresponding authors.Table S1. Predicted immunogenic peptides for the HLA class-IITable S2. Predicted immunogenic peptides for the HLA class-ITable S3. Immunogenicity assessment of the identified peptides using the VaxiJenThe authors declare no conflicts of interest.The authors are grateful to Amity University Lucknow Campus, UP, India, for providing the necessary facilities for this study. Also, the authors thank the Indian Council of Medical Research, New Delhi, India, for providing Research Associateship to Dr. Aupam Dhasmana (File No- 45/20/2013/NAN-BMS).


Kidney transplantation entails a high likelihood of endothelial injury. The endothelium is a target of choice for injury by ischemia-reperfusion, alloantibodies, and autoantibodies. A certain degree of ischemia-reperfusion injury inevitably occurs in the immediate posttransplant setting and can manifest as delayed graft function. Acute rejection episodes, whether T-cell or antibody-mediated, can involve the graft micro- and macrovasculature, leading to endothelial injury and adverse long-term consequences on graft function and survival. In turn, caspase-3 activation in injured and dying endothelial cells favors the release of extracellular vesicles (apoptotic bodies and apoptotic exosome-like vesicles) that further enhance autoantibody production, complement deposition, and microvascular rarefaction. In this review, we present the evidence for endothelial injury, its causes and long-term consequences on graft outcomes in the field of kidney transplantation. The endothelium plays an important role in vascular biology and regulation of renal function. Healthy endothelial cells are involved in vasodilation through nitric oxide (NO) release, which also inhibits platelet adhesion and aggregation, as well as leukocyte adhesion. Conversely, injured endothelial cells can develop a vasoconstrictive, pro-inflammatory, and procoagulant phenotype. Endothelial dysfunction is associated with traditional cardiovascular risk factors such as hypertension and diabetes, and it predicts atherosclerosis progression and cardiovascular events in the general population , 2. A laKidney transplantation is the best mode of renal replacement therapy, improving both quality of life and life expectancy compared to dialysis , 9. KidnLast, the most commonly used immunosuppressive agents may have divergent impact on the graft endothelium after transplantation. Mycophenolic acid may protect the endothelium, but calcineurin inhibitors have an adverse impact on endothelial function and glucocorticoids can worsen endothelial function under physiological conditions and improve it in the presence of inflammation. While these topics are reviewed elsewhere –14, hereThe kidney transplant procedure is inevitably associated with a certain degree of IRI. Donor type  and length of cold and warm ischemic times are important risk factors for IRI . ClinicaIn the past decade, microvascular injury and endothelial dysfunction have emerged as pivotal elements in the pathogenesis of AKI , 23. In Because the regenerative capacity of endothelial cells in peritubular capillaries appears limited –28, micrin vivo imaging and electron microscopy in murine models of AKI demonstrated a tight correlation between peritubular capillary injury, rarefaction, and renal fibrosis  molecules and prevents the development of an innate immune response to apoptotic cells . EffectoNevertheless, the impact of apoptosis may vary according to cell type and in certain conditions also favor inflammatory responses. Apoptotic endothelial cells externalize phosphatidylserine , which bin vitro and following transplantation in vivo. RIPK3−/− mice show better preservation of microvascular integrity in a model of cardiac rejection (The relative importance of regulated death pathways in AKI or rejection-induced microvascular injury is only beginning to be unraveled. It is generally accepted that broad caspase inhibition can prevent apoptosis at the expense of increased necroptosis and accentuated renal dysfunction , 78, a pejection . Whetherejection , 87. Alsejection , 89. Colejection , this tyEndothelial caspase-3 activation can promote vascular dysfunction through various and non-mutually exclusive pathways Figure . It favoKidney transplantation is associated with an elevated likelihood of damage to the graft macro- and microvasculature, given the IRI that occurs at the time of transplantation and the physical location of the graft endothelium that makes it a target of choice for cell- or antibody-mediated alloimmune injury. IRI, alloimmune damage, and autoantibodies can activate programmed cell death pathways in the graft endothelium, which can in turn trigger microvascular rarefaction, interstitial fibrosis, and graft dysfunction. These pathways represent potential targets for pharmacological intervention that could be delivered in preservation solutions during the period cold ischemia, with the aim of improving long-term graft outcomes.HC, MD, and M-JH reviewed the literature and drafted the manuscript.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.


Most of the inflammation in murine herpes simplex virus type 1 (HSV-1)-induced stromal keratitis (HSK) is due to exposure stress resulting from loss of corneal nerves and blink reflex. Corneal grafts often fail when placed on corneal beds with a history of HSK. We asked if corneal exposure contributes to the severe pathology of corneal grafts on HSV-1–infected corneal beds.Herpes simplex virus type 1–infected corneas were tested for blink reflex. Opacity and vascularization were monitored in allogeneic and syngeneic corneal grafts that were transplanted to corneal beds with no blink reflex or to those that retained blink reflex in at least one quadrant following infection.Retention of any level of blink reflex significantly reduced inflammation in HSV-1–infected corneas. Corneal allografts placed on HSV-1–infected beds lacking corneal blink reflex developed opacity faster and more frequently than those placed on infected beds that partially or completely retained blink reflex. Corneal grafts placed on infected corneal beds with no blink reflex rapidly became opaque to a level that would be considered rejection. However, protecting these grafts from exposure by tarsorrhaphy prevented or reversed the opacity in both syngeneic and allogenic grafts.Exposure due to HSV-1–engendered hypoesthesia causes rapid, severe, persistent, but reversible opacification of both allogeneic and syngeneic corneal grafts. This opacity should not be interpreted as immunologic rejection. Exposure stress may contribute to the high rate of corneal graft pathology in patients with recurrent HSK. In a 2002 perspective paper,9 the authors state that graft rejection in the mouse and rat models can be compared by simply grading the graft opacity level. However, the authors qualify the statement with the caveat that transient opacification of corneal grafts referred to as “rejection episodes” can occur and limit the reliability of graft opacity as the sole criterion for rejection.9 Although it was demonstrated that changing surgical techniques could reduce the incidence of these rejection episodes, the conclusion was that no good explanation for this phenomenon was available. Thus, there is a need for a more thorough understanding of the mechanisms of graft opacity in the mouse corneal transplantation model.Corneal transplantation has long been the most commonly performed transplant surgery in the United States and Great Britain.10 Herpes simplex virus-1 corneal infections result in leukocytic infiltration that can be subclinical or manifest as herpes stromal keratitis (HSK), an immunopathological process that represents the leading infectious cause of blindness in the United States.11 During primary HSV-1 infections, the virus establishes a latent infection in sensory ganglia that upon reactivation can serve as a source of virus for recurrent bouts of HSK. Thus, one could hypothesize that the high rate of sustained corneal graft opacity on HSV-1–infected corneal beds might result from: (1) residual inflammation and vascularization in the recipient cornea at the time of surgery, and/or (2) reactivation of latent virus and shedding at the corneal surface resulting in direct viral cytopathic damage or induction of damaging inflammation in the corneal graft. Indeed, evidence in support of both of these mechanisms has been reported.1217Here, we use corneal herpes simplex virus-1 (HSV-1) infections to create a physiologically relevant high-risk model of corneal graft rejection. Corneal HSV-1 infections are both a major reason for corneal transplants, and a major cause of corneal transplant rejection.1821 Recent findings demonstrate that the HSV-1–induced loss of corneal sensation is associated with partial or complete loss of corneal sensory nerves,2123 and manifests as a loss of blink reflex and corneal exposure and desiccation.22 Indeed, it was shown that most of the inflammation associated with HSK resulted from corneal exposure and could be largely prevented or resolved by performing a protective tarsorrhaphy.22 Interestingly, mice developed severe HSK subsequent to a complete loss of corneal blink reflex.22 Although corneal grafts inherently lack sensation due to severing of corneal nerves during excision, sensation is normally retained in the corneal bed. In contrast, when placed on an HSV-1–infected bed both the corneal graft and the surrounding corneal bed might lack sensation leading to exposure-induced inflammation in the corneal graft. These findings led us to hypothesize that HSV-1–induced corneal exposure might constitute an additional cause for the development of sustained opacity of corneal grafts placed on HSV-1–infected corneal beds. Our findings support this hypothesis. Furthermore, we demonstrate that in order to correlate persistent opacity with graft rejection, one must consider a contribution of long-term exposure stress to graft opacity.An association of HSK and other viral infections of the cornea with a loss of corneal sensation and exposure stress is well-established in humans.Female BALB/c and C57BL/6 mice purchased from Jackson Laboratories  were housed in the Animal Resource Facility at the University of Pittsburgh Medical Center . The use of animals was in accordance with the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research, and all procedures were approved by the University of Pittsburgh Institutional Animal Care and Use committee.4 plaque forming units (pfu) of HSV-1 strain KOS. Briefly, the mice were anesthetized with a mixture of ketamine and xylazine  at doses 150 and 14 mg/kg of body weight, respectively. Anesthesia was delivered to the mice via intraperitoneal injection using a 30-G needle. Mice then received unilateral corneal scarification and topical virus application. The scratches on the corneal epithelium were deep enough to reach but not penetrate the basement membrane.The scarified corneas of female 6- to 12-week-old BALB/c mice were infected by topically applying 1 × 10Tarsorrhaphy was performed under anesthesia using a surgical microscope . For infected but nontransplanted mice the procedure was performed after the skin disease and belapheratitis resulting from HSV-1 infection had resolved . Three mattress stitches were made with 8-0 silk suture  through the interstitial part of the eyelids after both edges of the eyelids were mildly cut with scissors. The adhesion of the eyelids prevented the mice from opening their eyes, protecting the corneal surface from exposure, and desiccation following the loss of corneal sensation. Tarsorrhaphy was also performed directly after some transplantations and maintained for periods of time dictated by the individual experiments.Allogeneic and syngeneic orthotopic corneal transplantations were performed on BALB/c mice. Donor corneas (2-mm diameter) from naïve C57BL/6 or BALB/c mice were excised and transplanted into the recipient cornea graft bed. The cornea graft was secured with eight interrupted 11-0 nylon sutures .For corneas without tarsorrhaphy, corneal opacities were scored at least twice a week in a masked fashion by two investigators with a high degree of consensus. Opacification was scored based on the following parameters: 0 = clear graft, 0.5 = small isolated imperfections or regions of translucence in the graft, 1 = minimal translucence or haziness extending over the entire graft with complete visualization of the iris and pupil, 1.5 = translucence or haziness extending over the entire graft and/or regional opacity in a minority of the graft, 2 = stromal opacity over the majority of the graft with visualization of the pupil margin and the iris structures, 2.5 = stromal opacity extending over the majority of the graft with only small regional translucence allowing very limited and strongly obscured visualization of the iris, 3 = complete stromal opacity with no visualization of the iris structures, 3.5 = complete stromal opacity associated with associated pathology such a granuloma or ulcer in the graft, 4 = destruction of the graft, a loss of graft integrity or graft perforation. Opacity associated with HSV-1 infections was assessed using the same scale and method used for measuring sustained graft opacity, but considered the whole cornea rather than just the graft. For the survival curves a graft was recorded as opaque when two consecutive scores of 3 or greater were observed. The incidence of sustained opacity was then recorded as the date of the first opacity score of 3 or greater. Measurement of neovascularization included the entire cornea, not just the graft, and the same scoring system was used for both transplanted and nontransplanted corneas. Briefly, the cornea was visually divided into four quadrants, and each quadrant was scored as 0 = no vessels visible, 1= vessels extend into the corneal bed, 2 = vessels extending up to graft interface or 50% of the distance to the center of the cornea, 3 = vessels extending into the graft but not reaching the center of the graft or extend 50% to 75% of the distance to the graft center, and 4 = vessels extending to the central cornea. The scores for all four quadrants were added to achieve a final score on a scale 0 to 16.The corneal blink reflex was tested and recorded as positive or negative by loosely holding the mouse and touching four areas in the periphery of the cornea and the center of the cornea with a sterile plastic probe being careful to avoid touching the eyelashes and whiskers see , S2. A lGroups of mice received HSV-1 corneal infections and TGs were excised following the resolution of skin disease and blepharitis (>15 dpi). The TGs were placed into a DNA preserving tissue digestion buffer purchased from Qiagen . The tissue was digested in the presence proteinase K and the DNA was isolated using Qiagen DNAeasy kit as per manufactures instruction. The viral copy number was assessed by using a real-time PCR protocol employing a labelled probe (5′-(FAM)TCCGGACCACTTTTC(NFQ)-3′), which recognizes the gene for glycoprotein H (gH). The reaction was carried out using TaqMan Universal PCR Master Mix . Because HSV-1 contains a single copy of gH, we compared the Ct values with that of a plasmid standard and then used a liner regression to calculate copy number. Mathematical and statistical analysis was performed using Applied Biosystems StepOne software  and GraphPad Prism Software .Splenocytes from C57BL/6 or BALB/c mice were exposed to 3000 rad of gamma radiation and used as stimulator cells. The graft recipient splenocytes were then incubated in triplicate cultures with an equal number of C57BL/6 stimulator cells , BALB/c stimulator cells (negative control), or BALB/c stimulator cells plus 0.1 μg/mL of anti-CD3  as a positive control. After 3 days in culture, the supernatants and the cells were harvested.2S04, and absorbance was read at 450 nm using a Synergy 2 plate reader .The supernatants were analyzed by sandwich ELISA for IFN-γ. Briefly, Immulon 4HBX high binding polystyrene ELSIA plates  were coated with anti–IFN-γ (clone R4-6A2) over night at 4°C, washed with PBS + 0.05% tween 20 , and the wells were blocked for 1 hour with PBS + 0.05% tween and 1% BSA (Sigma-Aldrich Corp.). The wells were washed and then loaded with either 100 μL of either diluted supernatant from the lymphocyte cultures, or of various concentrations of a purified mouse IFN-γ standard. The plates were incubated overnight at 4°C, the wells were washed and loaded with biotin conjugated anti–IFN-γ . After 1 hour at room temperature the wells were washed, loaded with streptavidin conjugated to horseradish peroxidase, and incubated for 30 minutes at room temperature. The wells were washed and loaded with 100-μL TMB peroxidase substrate . Color development was stopped by addition of 50 μL of 1N H5 pfu of HSV-1 KOS induces severe vascularization, opacity, and complete loss of corneal nerves and corneal sensitivity as assessed by blink reflex in nearly 100% of mice.22 A lower dose of the same virus was shown to induce severe HSK in only 50% of mice, but the relationship between loss of corneal sensitivity and development of HSK was not investigated in that study.24 Therefore, BALB/c mice received corneal infections with a suboptimal (1 × 104 pfu) dose of HSV-1 KOS. At 15 to 28 days post infection (dpi) corneas were tested for blink reflex by touching four quadrants of the peripheral cornea and the central cornea  were placed on HSV-1–infected corneal beds that completely lost blink reflex or maintained some level of blink reflex, or on noninfected corneal beds. Graft opacity was measured 9 days post transplant and representative photographs are provided A. FourteA sustained opacity score greater than or equal to 3 is a frequently used metric for defining immunologic rejection in mice. When placed on HSV-1–infected corneal graft beds that completely lost blink reflex, 95% of corneal allografts achieved a sustained opacity score greater than or equal to 3 within 11 days D. In con22 This inflammation is largely reversible by protecting the corneal surface with tarsorrhaphy.22 We hypothesized that exposure contributes to the severe opacity in allografts that are placed on corneal beds that have lost blink reflex. To investigate the contribution of exposure to graft opacity we placed corneal allografts (C57BL/6 donor to BALB/c recipient) or syngeneic grafts (BALB/c mice donors to BALB/c recipients) on HSV-1–infected corneas that completely lost blink reflex and had completely vascularized corneas. The mice were then divided into two groups. Tarsorrhaphy was performed on one group to prevent graft exposure. Graft opacity for the first group was measured at 9 days post transplant  and severe inflammation as measured by corneal opacity. In contrast, partial loss of corneal sensation (retention of blink reflex in one or more sectors of the cornea) was not associated with increased corneal inflammation .First, our earlier work demonstrated that induction of HSK is dose-dependent in BALB/c mice, with 100% disease incidence at an infectious dose of 1 × 10ammation . Our pre29 It is unclear if the corneal nerve degeneration is caused by inflammation at nerve termini, virus infection of the nerves, or a combination of both. Our observation that mice that completely lose cornea sensitivity have similar mean TG genome copy number to those with partial sensory loss is consistent with an inflammatory rather than a viral etiology (Herpes simplex virus type 1 corneal infection results in rapid infiltration of leukocytes into the cornea and also viral invasion of sensory nerves and transmission to the TG. The pathology seen in mice resembles neurotropic keratitis, which can be caused by damage to TG.etiology .30 Because both syngeneic and allogeneic grafts rapidly developed severe and sustained opacity, we concluded in that work that the inflammation resulted from an innate immune response emanating from the inflamed recipient corneal bed. Here, we have amended those findings. There is innate inflammation driving the rapid opacification of cornea grafts, but the driver of that inflammation is exposures stress. Thus, mice that retain blink reflex develop less severe opacity during the first 9 days post transplant (Our previous findings reported that both syngeneic and allogeneic corneal grafts rapidly developed opacity when placed on HSV-1–infected corneal beds that exhibit severe HSK-associated inflammation.ansplant , and proansplant . Perhaps31 The tarsorrhaphy does more than just protect from exposure; it also stops tear film drainage.32 Thus, the dramatic recovery we see in mice the received a tarsorrhaphy may be due to prolonged contact with components of tear film and lid mucosa as well the removal of the exposure stress.Our study demonstrates that the enhanced sustained opacity in corneas transplanted to HSV-1–infected corneal beds can be reversed by tarsorrhaphy, a standard surgical procedure performed in animals and humans to prevent corneal exposure.Corneal transplantation eliminates sensitivity in the transplanted corneal button. However, our data emphasize the importance of retaining sensitivity in the surrounding corneal bed. If the corneal bed completely lacks sensitivity, the transplanted cornea will become rapidly opaque due to exposure keratopathy. This keratopathy is largely avoided if sensitivity is retained in even a single quadrant of the corneal bed or if the corneal bed is protected by tarsorrhaphy following transplant. Thus, assessments of sensitivity in the corneal bed should be included in any transplant model. In humans, consensual blink reflex might mitigate some or all of the impact of grafting to a graft bed that lacks sensation, but a possible contribution should be considered.Supplement 1Click here for additional data file.Supplement 2Click here for additional data file.Supplement 3Click here for additional data file.


Thermal studies showed that the thermal stability and crystallization temperature of the PUS composites increased compared to that of PU. The loss modulus curves showed that neat PU crystallizes at 124 °C and at 127 °C for PUS-0.5 wt.% and rises with increasing loading from 0.5 to 2 wt.%.The advancements in material science and technology have made polyurethane (PU) one of the most important renewable polymers. Enhancing the physio-chemical and mechanical properties of PU has become the theme of this and many other studies. One of these enhancements was carried out by adding starch to PU to form new renewable materials called polyurethane-starch composites (PUS). In this study, PUS composites are prepared by adding starch at 0.5, 1.0, 1.5, and 2.0 wt.% to a PU matrix. The mechanical, thermal, and morphological properties of PU and PUS composites were investigated. Scanning electron microscope (SEM) images of PU and PUS fractured surfaces show cracks and agglomeration in PUS at 1.5 wt.% starch. The thermo-mechanical properties of the PUS composites were improved as starch content increased to 1.5 wt.% and declined by more starch loading. Despite this reduction, the mechanical properties were still better than that of neat PU. The mechanical strength increased as starch content increased to 1.5 wt.%. The tensile, flexural, and impact strengths of the PUS composites were found to be 9.62 MPa, 126.04 MPa, and 12.87 × 10 Starch (amylum) is a natural renewable polysaccharide polymer obtained from a great variety of crops and is a promising raw material for producing biodegradable products ,2,3. StaPolyurethanes (PUs or PURs) are used as host polymers for many applications. PU is composed of organic units called urethane and appears in two types: thermoset and thermoplastic polymer ,9. PUs aDuring the past 40 years, there have been numerous studies and excellent publications on the preparation and the structural, thermal, mechanical, and morphological characterization of TPU systems by various research groups ,12. PUs In this report, PU and PUS composites at different percentages of starch additive were prepared and investigated. The investigation included mechanical properties, physical appearance and morphology, storage capacity, and thermal properties. The study adds very important results regarding the mechanical properties which could enhance the applications of these new composites.The morphologies of the PU surface and starch powder are shown in The SEM images of the tensile fractured surface of PUS composites at different starch loadings are shown in The SEM images of the impact fractured surfaces of PUS composites at different starch loadings are shown in The results of the tensile strength of neat PU, PUS-0.5%, PUS-1.0%, PUS-1.5%, and PUS-2.0% are shown in The Young’s modulus was then tested for neat PU and PUS composites as presented in The maximum load beyond the yield that PU or PUS composites can hold before breaking down is shown in The elongation at break measurements shown in Applying a load in the middle of a sample that is supported at its ends demonstrates a material’s maximum compressive stress and maximum tensile values at the supporting points. The basic requirement of a successful flexural test is that the sample has to withstand maximum tensile strength before failing under compressive strength; otherwise, there is no reason to conduct the flexural test. The flexural test of the neat PU and PUS composites at different starch loading is shown in The maximum load at the yield of the neat PU and PUS composites is shown in The impact test is expressed by the energy or, more specifically, the intensity which is the power per surface area. The basic condition for the impact test is to fracture the specimen by evenly distributing enough load throughout the sample. There are two impact tests; one without a notch and the other with a notch, where the notch is a groove made by very sharp razor with specifications determined by a standard based on the machine itself.−3 J/mm2 and as starch loading increases, the impact factor increases showing its maximum at 12.87 × 10−3 J/mm2 at 1.5 wt.% starch loading. The results show an improvement of about 103% compared to the neat PU which, by increasing starch loading, declines to only 38% at the highest starch loading of 2.0 wt.%. Apparently, the morphology of the higher starch loading samples was distorted based on SEM analysis.The impact strength test of un-notched PU and PUS composites were carried out and the results are shown in −3 J/mm2, declining by 47% compared to the original un-notched sample. The impact strength increases as starch loading percentage increases for the composites samples and reaches its maximum at 6.33 × 10−3 J/mm2 and, similarly to un-notched samples, declines for higher starch loading contents.The last mechanical test is shown in The effect of notching the sample could be used to show that the smoothness of the sample surfaces plays an important role in the impact strength . The resFor completeness, the results presented in this section are summarized in gT) temperatures, storage and loss moduli, and heat deflection or softening points.The DMA technique provides viscoelastic measurements of the response of a material under the influence of a sinusoidal oscillation force, which might be in the form of a strain (strain controlled instrument) or a stress (stress controlled instrument). The stress/strain could be in any mechanical form such as tension, compression, or torsion. For a perfectly elastic solid, the strain and the stress are in phase, while in a purely viscous media, the strain lags the stress by 90°. The phase difference lies somewhere between 0° and 90° for all other solid materials depending on the rigidity and elasticity. The storage modulus (cT) of PU. Adding starch to a PU matrix enhances the nucleation process of PU crystallization by increasing the surface area of the PUS and providing more nucleation sites for the PU to crystallize.The loss modulus (mT). The results show that mT shifts from 156 to 150 °C for PU-2.0 wt.% starch.The TGA curves for neat PU and PUS composites at different loadings are shown in The degradation at d out by . Figure There are only two materials included in this paper: thermostat PU and natural starch in powder form. The most important properties of PU and starch are listed in The sample preparation and the experimental procedures were conducted according to the flow chart shown in E′) and loss modulus (E′′). DMA spectra were taken in tensile mode at 1 Hz frequency in a broad temperature range (50 to 150 °C) with a programmed heating rate of 2 °C/min using a Dual cantilever method, the force applied was 6.35 N, displacement 20 μm. Finally, thermal stability and composition were assessed using Thermogravimetric  . Samples of about 7–8 mg each were heated from 50 to 950 °C at a rate of 10 °C/min under nitrogen atmosphere.The morphologies of the composites’ fracture surfaces were studied using a scanning electron microscope (SEM), Hitachi TM 3000, Somerset, NJ, USA. The 3-D SEM images are, therefore, useful for judging the surface structure of the sample. Tensile properties were determined using an Instron universal testing machine , with a 200 Newton load transducer, according to the ASTM D-638 type V method standard . The tenImproving the mechanical and thermal properties of PU is very important due to an essential demand for industrial and technological use. The thermo-mechanical properties are improved by adding starch to PU at very small amount of 0.5, 1.0, 1.5, and 2.0 wt.%. The effect of loading starch was tested thermally and mechanically by several techniques, such SEM, tensile strength, elongation, Young modulus, maximum load at yield, and DMA of PU and PUS. SEM images of the fracture surface have shown two features: the formation of agglomerated starch over the surface and the potentiality of PUS composites to having cracks at about 1.5 wt.% starch loading. The mechanical properties were improved by 17% for the tensile strength, and about 91% for maximum load at the yield before being adversely reduced due to the agglomeration and cracks. Even the mechanical properties declined after 1.5 wt.% starch loading, however, the mechanical properties never returned the PUS composites to the standard of PU alone. To show the effect of the natural cracks, a test was performed to investigate the effect of creating notches on the PUS surface. The result showed a significant effect of such an artificial crack by reducing impact strength by 103% compared with a non-notched sample. This result shows the importance of the smoothness of the surface, as well as the prevention of the formation of cracks. DMA analyses of storage and loss modulus have shown that the crystalline temperature and melting temperature were affected. These results may lead to expanding the use of PUS composites.


Incompatible coefficient of thermal expansion (CTE) is supposed to be a reason for chipping of ceramic veneered zirconia. This study evaluates the effect of veneering ceramic at varied CTE on bond strength to zirconia.Zirconia disks  were prepared from Y-TZP (Cercon®) and sintered at 1350°C for 6 hours. All zirconia disks were veneered with ceramics  with varied CTE including VITADur® alpha (VDα), VITAVM®7 (VM7), VITAVM®9 (VM9), Cercon® ceramkiss (CCK), IPSe.max® ceram (IeC), and IPS dSIGN® (IdS) (n=15). The specimens were thermo-cycled  prior to determine the shear bond strength on a universal testing machine. The veneering ceramic and zirconia rods  were prepared for CTE evaluation. ANOVA and Tukey’s multiple comparisons were used to determine the statistically significant difference (α=0.05). Weibull analysis was applied for survival probability, Weibull modulus (m), and characteristics strength (σo) of the shear bond. The interfaces were microscopically examined. The phase transformation of zirconia was determined using X ray diffraction.p<0.05). The t→m phase transformation related with the CTE difference. The mean±sd (MPa), m, and σo of bond strength were 20.45±2.32, 9.25, and 21.53 for Z-VDα, 19.47±4.53, 4.66, and 20.31 for Z-VM7, 21.05±3.96, 5.61, and 21.88 for Z-IeC, 25.85±2.74, 9.93, and 27.15 for Z-VM9, 25.82±4.39, 6.27, and 27.06 for Z-CCK, and 2.96±0.73, 4.11, and 3.28 for Z-IdS. The CTE (×10-6/°C) were 10.80, 7.83, 7.87, 9.86, 9.93, 10.03, and 12.95 for Z, VDα, VM7, IeC, VM9, CCK, and IdS. The bond strength was significantly affected by the CTE difference (The CTE’s differences induced stress that affected the bond strength. CTE’s compatibility of veneering ceramic to zirconia is crucial for enhancing the bond strength. The CTE difference approximately 0.77-0.87×10-6/°C was recommended. Key words:Bond strength, coefficient of thermal expansion, zirconia. The celebrity of using all ceramic restoration has been increasing as they provide highly desirable esthetic appearance and extreme biological compatibility. Patients often request for metal-free restoration, which makes ceramic the most preferred restoration choice for reconstruction. Several new dental ceramics have been developed with improved strengths to withstand the stress from the physiologic masticatory function, which are being used as long-span fixed prostheses . Among cThe bond firmness between zirconia and ceramic that is partly related with the compressive force generated is because of appropriate difference of CTE’s value of veneering ceramic and zirconia -26. The The shear bond strengths of six ceramics, based on different CTE, including VITADur® alpha , VITAVM®7 , VITAVM®9 , Cercon®ceramkiss , IPSe.max® ceram , and IPS dSIGN®  veneered to Cercon® zirconia  were evaluated.-Zirconia Specimen PreparationNinety zirconia disks (12.0 mm in diameter and 1.2 mm in thickness) were prepared into disk shape from partially sintered Y-TZP blank using precision machine , and sintered in the furnace  at the recommended temperature of 1350ºC for 6 hours, to derive fully sintered zirconia disks (10.0 mm in diameter and 1.0 mm in thickness) due to 20% volumetric sintering shrinkage.-Ceramic Veneering TechniqueAll zirconia disks were randomly divided into six groups (15 disks per group) to be veneered with VDα, VM7, VM9, CCK, IeC, and IdS ceramic . The zir-Thermal Cycle TechniqueThe specimens were subjected to thermo-cycle process for 500 cycles between 5°C and 55°C for 30 seconds of immersion in each temperature bath, and 5 seconds for specimen transfer prior to the assessment of the shear bond strength.-Evaluation for Shear Bond StrengthEach specimen was evaluated for its shear bond strength by subjecting it to a compression shear apparatus on a universal testing machine . The shear load was applied at the zirconia-ceramic interface at crosshead speed of 0.5 mm/min. The loads at failure were recorded and calculated for shear bond strength  by dividing the failure load  by the area of interface , as shown in Equation 1: σ=P/A ……………….Equation 1.-Determination of Coefficient of Thermal ExpansionThe veneering ceramics and zirconia were prepared into a round rod shaped specimens . The creamy mixed consistency of veneering ceramic was placed, condensed into a silicone mold, and then removed for firing and glazing process. The zirconia rod was prepared from partially sintered Y-TZP blank and sintered in the furnace to derive the final dimension. The horizontal single rod dilatometer  was used to determine the heating and cooling curve for each specimen from 25°C to 500°C at the rate of 5°C per minute. The linear CTE was determined by using Proteus® software version 4.7 .Microscopic ExaminationThe de-bonding surfaces were examined under optical stereomicroscope  and scanning electron microscope  for characterizing the bonding failure’s mode. The zirconia-ceramic interface for each group was examined with SEM, and energy dispersive x-ray spectroscopy .-Evaluation of Crystalline StructureThe crystalline phases of ceramic veneered zirconia was determined for the relative amount of monoclinic (m) and tetragonal (t) phase of Y-TZP using the X-ray diffraction . The specimens were scanned with copper k-alpha (Cu Kα) radiation from the 2θ degree of 20–40o with 0.02o step size at every 2 seconds’ interval. The phase was analyzed in comparison to the known standard database of the joint committee on powder diffraction standards (JCPDS), and calculated for corresponding d-values using Bragg, as shown in Equation 2 : λ = d2 Where: λ is the X-ray wavelength (0.15418 nm for CuKa), d is normal distance of planes with the Miller indices (hkl), and θ is the Bragg angle.The ratio of m- to t- phase was determined by the peaks’ intensities using X’Pert Plus software . The mass fraction of m-phase to the total zirconia phase content was calculated from Garvie-Nicholson formula, as shown in Equation 3, and further corrected for nonlinearity using Toraya formula, as shown in Equations 4 and 5 , Fig. 11.Where: Im and It: integral intensities of monoclinic and tetragonal phaseC: composition-dependent correction factor (C = 1.32)Xt and Xm: the Toraya-corrected mass fraction of tetragonal and monolithic zirconia-Statistical Analysis of DataThe data was statistically analyzed using SPSS/PC Version 20 software . An analysis of variance (ANOVA) was used to determine the significant differences of shear bond strength upon different ceramics. Post-hoc Tukey HSD multiple comparison was used to determine the difference between groups at 95% level of confidence. Weibull analysis was performed to determine the bond strength’s reliability using Weibull++®statistics , and estimated the Weibull modulus (m) from Equation 6 and from a slope of the straight line plotted between ln{ln(1/Ps(Vo)} against m ln(σ/σo). Ps (VΟ)=exp{-(σ⁄σΟ)m } …………..Equation 6Where: Ps (Vo) is the probability of survival as the fraction of identical sample; Vo is the volume of the sample;σ is the shear strength; σo is the Weibull characteristic strength; and m is Weibull modulus.p<0.05) (p<0.05), except for the groups of Z-VM7, Z-VDα, and Z-IeC (p>0.05)  . Post-ho(p>0.05) . Weibull(p>0.05) B,D,F,H,JThe x-ray diffraction patterns revealed that most of crystal structure of tetragonal (t) phase with minor amount of monoclinic (m) phase in every group as shown in Figure This study indicated that bond strength of ceramic veneered zirconia is affected by veneering ceramics that possess different CTE values. Thus, null hypothesis was rejected. The CTE displays a major role in bond strength of ceramic veneered zirconia ,18. The The CTE’s difference principally influenced the interfacial residual stress that reflected on the amount of t→m phase transformation of zirconia . The resTo generate suitable levels of residual stress for promptly bond strength, appropriate CTE difference need to be primarily considered . This stThis investigation described the role of CTE in bond strength of ceramic veneered to Y-TZP. The study proved that bond strength of ceramic veneered zirconia was influenced by the CTE’s difference between zirconia and veneering ceramic. An extreme CTE difference between veneering ceramic and zirconia did not render favorable bond, but resulted in a higher stress inducing phase transformation of zirconia to facilitate the ease of bond failure. Proper selection of veneering ceramic based on appropriated CTE difference is extremely crucial to assure durable bond strength of ceramic fused with zirconia. The CTE of veneering ceramic less than that of zirconia for approximately 0.77-0.87X/ oC was suggested to provide suitable residual compressive stress for conferring a favorable bond strength.


The effect of non-ionizing microwave radiation on starch is due to a gelatinization temperature range that changes starch structure and properties. However, the changes in starch upon microwave heating are observable throughout the heating process. We compared the effects on starch heating by microwaves to the effects by rapid and regular conventional heating. Our results show that microwave heating promotes the rapid rearrangement of starch molecules at low temperatures; starch showed a stable dielectric response and a high dielectric constant. Microwave heating changed the Cole-Cole curve and the polarization of starch suspension at low temperatures. A marked transition at 2.45 GHz resulted in a double-polarization phenomenon. At temperatures below gelatinization, microwave-induced dielectric rearrangement and changes in the polarization characteristics of starch suspensions reduced the absorption properties; at temperatures above gelatinization, these characteristics became consistent with conventional heating. Throughout the heating process, microwaves change the electrical response and polarization characteristics of the starch at low temperatures, but on the macro level, there is no enhancement of the material’s microwave absorption properties. In contrast, with the warming process, the starch exhibited a “blocking effect”, and the absorption properties of the starch quickly returned to the level observed in conductive heating after gelatinization. These reports confirmed that microwave heating causes a rearrangement of intramolecular structures during gelatinization, thus altering the water absorption capacity, solubility, swelling ability, gelatinization characteristics, dehydration shrinkage behavior and rheological properties of starch5. We have long studied the relationship between gelatinization and the heating rate. The key factor that influences the physicochemical behavior of starch under microwave irradiation is the rapid heating effect; the thermophysical and structural properties are similar when conduction heating is carried out at the same rate as microwave heating11.Microwave radiation is non-ionizing and changes the structural characteristics of materials during processing. It is one of the physical means used to modify the structural characteristics of starch. To promote the understanding and application of the physical effects of this process, MuzimbarandaThe effect of microwaves on materials differs from heat transfer in that microwave energy transfer is instantaneously stable and behaves like an excited electromagnetic wave. Some researchers have hypothesized that a microscopic response is instantly generated in material exposed to a microwave field; macroscopic changes may arise from an accumulation of microscopic responses or from the summation of microscopic responses together with other variables. Alternatively, a microscopic response may be too small to cause significant effects on the system. Traditional physical and chemical methods cannot verify or invalidate the above assumptions. We have found that electromagnetic and statistical methods can be used to preliminarily evaluate the characteristics of microwave absorption. In addition, we found that the absorption capability of starch differed in various gelatinization states. However, the causes for these differences and when they are generated are as yet unclear. As traditional methods cannot explain the above-mentioned process, it is necessary to use cross-disciplinary experimental and analytical methods to explore the electromagnetic nature of microwave radiation.13. The coupling efficiency between microwave radiation and polymeric material depends on the intensity, mobility, aggregation state and matrix state of the dipole14. The response of the dipole oscillation field consists of rotation and/or vibration, depending on the symmetry of the molecule; this molecular symmetry determines the degree of heat generation15. Based on this dipole mechanism, water is an important factor in the process of heating starch. Previous reports found that some low-wave properties of materials exhibited abnormal electromagnetic behavior in response to radiofrequency and microwave radiation after the materials were mixed with a strong dielectric material16. Starch is a carbohydrate polymer with a complex structure, composition and aggregation pattern. Pure liquid water absorbs microwave energy across a broad spectrum. Polymer matrix materials dispersed in water undergo different degrees of adsorption, binding and release upon heating; as a result, the material’s molecular polarization behavior changes and a power shift occurs in the dielectric response spectrum17.Polymers absorb microwaves by repositioning their dipoles in an applied electric field18. The Cole-Cole model is usually used to study the Debye relaxation behavior of monomeric materials to determine the polarization behavior through the semicircular curve of the first quadrant; this constitutes a low-field detection method in which the Cole-Cole model describes the polarization behavior of the dielectric19. Although researchers have used the Cole-Cole model to study the structural effects of high-field treatment on a single material, no study has yet applied this method to liquid mixed materials, especially for the study of liquid dispersions of food macromolecules.Earlier studies have shown that the Cole-Cole model is superior to the macroscopic Debye model for describing the dielectric relaxation behavior of pure waterWe evaluated the difference between microwave heating and traditional heating and established a microwave heating (MW) model based on rapid conduction heating (RCV) temperature. Using traditional conduction heating (CV) as a control method, we investigated MW, RCV, and CV. We used these methods to investigate the change in the dielectric properties of starch in a microwave field. We discuss here the Debye polarization mechanism of the corresponding system, as well as the relationship between the variation and the absorption characteristics of the system.Rice starch  was isolated and purified from fresh rice .8.A 30-g sample of fresh rice was steeped in 47 g distilled water for 18 h and ground into a slurry. The pH of the slurry was adjusted to 5.0 by adding 0.1 mol/L HCl; 0.04 g cellulase  was then added to the slurry, which was allowed to react at 50 °C for 2 h. The pH was then adjusted to 7.0 by adding 0.1 mol/L NaOH, and 0.9 g neutral protease  was added and allowed to react at 50 °C for 4 h. The slurry was then centrifuged at 10,000 g for 30 min. The soft top layer was carefully removed and the starch layer was reslurried and washed nine times with deionized water. Butyl alcohol was added to the starch solution, stirring for 18 h to remove the fat content. The isolated starch was then dried in a freeze dryer, passed through a 150-μm sieve and stored in a dryer at room temperature until analyzedA thermostat  with an oil bath (200 °C) was used for rapid heating. The starch suspensions (100 g) with a concentration of 6% (w/w) were prepared with distilled water and heated to the desired temperature in the oil bath under constant agitation while monitoring the temperature online using an optical fiber probe . The suspensions were then immediately placed into an ice bath to stop the gelatinization process. The treated samples were lyophilized and crushed to pass through a 75-μm sieve.The MV experiment was conducted in the same manner as described above, except a microwave oven was used for heating. A 2450-MHz oven  with an output power of 1.2 kW was used to match the heating rate of the conventional heating with an oil bath. Multiple replications were performed in the preliminary stages to determine the power necessary to match the oil bath.The CV experiment was conducted in the same manner as described above, except the sample mass was increased to 200 g and a hot plate was used for heating. Its measured heating rate was 0.13 °C/s.19. The probe was first calibrated using three materials , to ensure the accuracy of the measurements. Each sample was mixed to homogeneity before testing and was measured in triplicate.The dielectric properties of the liquid system were measured using a vector network analyzer  with an open-ended coaxial line, connected to a dielectric probe 12.The reflection loss (RL) versus frequency for complex liquid materials was improved on the basis of the arch method by adding polytetrafluoroethylene (PTFE) as the wall material to the former aluminum plate, to make a specialized container Fig. . The tesAll data generated or analysed during this study are included in this published article.7. However, we paid attention to the subgelatinization levels in this work, in which the highest heating temperature was 70 °C. The multiple correlation coefficient obtained from the function corrcoef in MATLAB was 0.9998. Therefore, the RCV model fitted well with the microwave processing  intermolecular hydrogen bonding in starch. In addition, swelling of the starch granules resulted in the expansion of bonded hydroxyl in the granule to form a hydrogen bond with HAt a microwave frequency of 2.45 GHz, a specific phenomenon was observed during the dielectric constant test of the starch emulsion: the heating mixture system’s ε″ was larger than any of the components. We examined next the behavior of the relaxation in the Debye model to investigate the causes of this specific phenomenon in the mixed system.30. Here we introduce the concept of complex permittivity. The real aspect of complex permittivity reflects the ability of the dielectric effect to store energy in the polarization process, whereas the imaginary aspect describes the energy dissipation during the polarization process. In the present study, three polarization mechanisms of the medium were ion polarization, electron polarization and electric dipole moment polarization. Ion polarization and electron polarization in the microwave frequency range are relatively weak, whereas they are more obvious in the infrared frequency range32. Therefore, we assume that the dipole moment polarization in the gelatinization process leads to a change in the dielectric loss of starch. In the process of polarization, microwave energy turned into heat, resulting in loss.The most basic physical concept of the dielectric effect is polarization, the offset of the positive and negative charge centers in the media. The establishment and change of polarization, known as the dielectric relaxation, occurs gradually over time as one equilibrium state transforms to another. In essence, the dielectric relaxation reflects the energy exchange and the related energy loss of microscopic particles during polarizations and ε∞ are static and optical frequency permittivities, respectively, ω is angular frequency, ω = 2πf and τ is relaxation time.The Debye equation introduces the concept of relaxation time, which describes dielectric relaxation as follows:ωτ in Equation , the material is a high-loss material that exhibits an interface polarization mechanism.ωε″) according to the test results of the real and imaginary part of dielectric constant under the limited frequency; thus, we can obtain parameters τ, ε∞ and εs according to the slope and intercept.a is the average relaxation time, α is a positive number between zero and 1, and α can be used to measure the applicability of the Debye parameters. The larger α is, the more the dielectric deviates from the Debye relaxation.In contrast, if we rewrite the Debye Equation  as shownEquation :8\documeα in the modified Debye equation.The Debye equation is a phenomenological theory that describes the complex energy exchange behavior of microscopic particles in the polarization process as attributed to relaxation time. Relaxation time τ is a function not only of composition and temperature, but also of the structure and microstructure of the material. However, the effect of the inhomogeneous microstructure of the material on the dielectric relaxation can be included in the value of 33.Finally, note that when the electric field frequency is very high (ω→∞), the Debye equation contains a ε′ → ε∞, ε → 0 function. In fact, the dielectric loss is still in the optical frequency , which is ignored in the Debye relaxation theory. Therefore, the Debye equation is limited because it is only suitable for a frequency range far below the optical frequency range0 (v)is the ordinate, and 1/2πf is the slope corresponding to the polarization frequency of Debye The simplified Cole-Cole equation is 0 . This reduction is due mostly to the absence of particulate (not hydrated) starch in a strong electric field. The charged particles cannot break through internal forces between water molecules. Because of the limit of the electrostatic charges on water molecules on the interface layer, at low starch concentrations the water-starch system generates a strong electromagnetic absorption interference. However, with an increase in temperature, starch granules absorb water and swell, resulting in a breakdown of lamellar starch structure and damage to entangled starch chains. During the hydration process, the positive and negative charges of the starch molecules do not coincide, so that the microscopic particles form an electric dipole, begin to exhibit polarity and produce polarization. In addition, as the degree of gelatinization of the system increases, there is considerable enhancement of the absorption characteristics of the materials.36. This experiment revealed an ion polarization contribution37, thus, double-polarization characteristics resulted from the combined action of multiple starch structures that antagonize the polarization process to a certain extent. Further research should help to elucidate these effects.After microwave treatment, the microwave absorption properties of the material did not increase under the experimental conditions, but showed a certain blocking effect in the heating process. We suggest three reasons for this blocking effect. First, combined with impact of the microwave on dielectric properties of the materials, the results showed that ε′, ε″ and tanδε of microwave-treated samples were higher than they were with the other two heating protocols before the gelatinization temperature was reached to cause the numerical increase in dielectric properties. Regarding electromagnetic characteristics, the greater the difference in dielectric constant and permeability, the easier it was to generate a blocking effect that influenced the absorption of electromagnetic waves. However, the starch aqueous permeability was about 1 with no marked magnetic effects; therefore, the microwave heating process amplifies the dielectric permeability difference and reduces microwave absorption. Second, although the electromagnetic loss of microwave-treated material is relatively increased, an impedance matching process of a two-phase system in the electromagnetic wave incident process was changed because the system exhibited a rearrangement of charge based on the material characteristics in the heating process. Therefore, microwave energy incident at the surface and interface was greatly reflected or offset, and that energy was not completely absorbed by the material. Third, combined with the effect of microwaves on the material Debye polarization mechanism, the results explain that although the microwave treatment at low temperatures changes the polarization characteristics of the material close to the frequency of 2.45 GHz, increasing the polarization process of the material, the dual-polarization characteristics of microwave treatment is related to the intrinsic polar moment of the polar molecules and the microelectronic polarization or atomic polarizationWith microwave heating, starch exhibits a relatively stable electromagnetic response before and after gelatinization. The starch suspension rearranges at low temperatures, inducing standardization of the electric dipole moment in its main components and forming a more stable polarization relaxation behavior; in addition, we observe more prominent and stable dielectric properties. At the microcosmic level, microwave heating has a marked effect on the Cole-Cole reduction curve of starch suspensions, changing the original polarization and producing a double-polarization phenomenon that affects the microwave absorption based on the polarization mechanism. At the macroscopic level, microwave heating hinders the microwave-absorbing properties of starch suspensions below the gelatinization temperature; this is consistent with the conduction heating method following gelatinization.We could observe effects of microwaves on starch and the production of starch suspensions throughout the heating process. During the early stage of heating, microwaves immediately affected the dielectric response and the polarization mechanism of the starch. During the middle and late stages of heating, the microwave-absorbing properties under a microwave field markedly changed with the thermal conversion process. This provides a powerful basis for further explanation of microwave thermal runaway.


Pharmaceuticals improved its ranking [Pharmaceuticals confirms its position in the first quartile in the category of pharmaceutical science. Moreover, papers published in our journal in 2013 and 2014 have been cited 4.5 times, on average, in 2015.As explained in the editorial written 12 months ago [It is my pleasure to confirm the progress recorded in 2016. Indeed, the number of published manuscripts has jumped from 45 to 79 in 2016 volume. This includes submissions received in 10 completed Special Issues. They were dedicated to the First International Electronic Conference on Medicinal Chemistry  , Thierry Besson , Francois Dufrasne , Marc Le Borgne , Andrea Porcheddu , Osvaldo A. Santos-Filho , Gary J. Stephens , and Nigel Yarlett .Pharmaceuticals confirmed its status as an international journal by being present as a media partner, an exhibitor, or a sponsor at national and international meetings among which were:Biosensors 2016, Twenty-sixth Anniversary World Congress on Biosensors, 25–27 May, Gothenburg, Sweden;Thirtieth France-Belgium Meeting of Pharmacochemistry, 25–27 May, Amboise, France;Aptamers in Bordeaux 2016, 24–25 June, France;CLINAM 9/2016 Conference and Exhibition, European and Global Summit for Cutting-Edge Medicine: Clinical Nanomedicine and Targeted Medicine, Enabling Technologies for Personalized Medicine, 26–29 June, Basel, Switzerland;ACS National Meeting and Exhibition, 21–23 August, Philadelphia, PA, USA;Réunion du Groupement des Pharmacochimistes de l’Arc Atlantique, 25–26 August, Angers, France;Eighth International Conference on Protein Kinase CK2, 6–9 September, Homburg, Germany;Thirteenth European Biological Inorganic Chemistry (EuroBIC 13) Conference, 28 August–1 September, Budapest, Hungary;Twenty-fourth National Meeting in Medicinal Chemistry, 11–14 September, Perugia, Italy.Fourth Conference on Innovation in Drug Delivery: Site-Specific Drug Delivery, 25–28 September, Antibes-Juan-les-Pins, France;Pharmaceuticals was the organizer of the Second International Electronic Conference on Medicinal Chemistry [hemistry . The evehemistry , submittPharmaceuticals Travel Award for Young Investigators [Mentions should be made, in addition, that we received 83 outstanding applications for the 2016 Pharmaceuticals will offer a travel grant to help support a young investigator in her/his professional career. Details of the competition can be found on our website [http://sciforum.net/conference/ecmc-3.In 2017, we shall pursue our efforts to raise the journal to a better position still and to increase its visibility. Therefore, we can already announce that It is my pleasure to end this editorial by wishing you a healthy and prosperous new year. This is also the opportunity for me to warmly thank, for their confidence, our authors, readers, and reviewers, as well as the scientists of the editorial board, members of our teams in Basel, Barcelona, Beijing, and Wuhan, and our sponsors.