We compared ex vivo activation of CD8+ T lymphocytes from HIV-infected individuals by wild type HIV peptide epitopes and synthetic variants bearing simulated A3G/F-induced mutations by measuring interferon-γ (IFN-γ) production. We found that A3G/F-induced mutations consistently diminished HIV-specific CD8+ T cell responses against the common epitopes we tested. If this reflects a significant trend in vivo, then adaptation by HIV to enrich sequences that are favored for mutation by A3G/F (A3G/F hotspots) in portions of its genome that encode immunogenic CD8+ T cell epitopes would favor CTL escape. Indeed, we found the most frequently mutated A3G motif (CCC) is enriched up to 6-fold within viral genomic sequences encoding immunodominant CD8+ T cell epitopes in Gag, Pol and Nef. Within each gene, A3G/F hotspots are more abundant in sequences encoding epitopes that are commonly recognized due to their HLA restriction. Thus, in our system, mutations of the HIV genome, mimicking A3G/F activity, appeared to abrogate or severely reduce CTL recognition. We suggest that the physiological significance of this potential effect in facilitating CTL escape is echoed in the adaptation of the HIV genome to enrich A3G/F hotspots in sequences encoding CTL epitopes that are more immunogenic at the population level.Due to constitutive expression in cells targeted by human immunodeficiency virus (HIV), and immediate mode of viral restriction upon HIV entry into the host cell, APOBEC3G (A3G) and APOBEC3F (A3F) have been considered primarily as agents of innate immunity. Recent bioinformatic and mouse model studies hint at the possibility that mutation of the HIV genome by these enzymes may also affect adaptive immunity but whether this occurs in HIV-infected individuals has not been examined. We evaluated whether APOBEC-mediated mutations within common HIV CD8 They presumably play a particularly important role in the early phases of HIV infection to limit viral propagation whilst long-lasting and more specific adaptive immune responses takes weeks to months to develop fully.In the physiological setting, the effectiveness of A3G/F against HIV is restricted by several factors. First, the HIV auxiliary protein viral infectivity factor (Vif) binds to A3G/F and promotes their degradation Following the initial innate immune barriers, adaptive immune responses in the form of anti-viral antibodies and cytotoxic T cells (CTL) develop and act to contain viral propagation throughout the course of infection + T cells of HLA-matched, HIV-infected individuals ex vivo. We found that HIV-specific CD8+ T cell recognition was generally diminished by A3G/F-induced mutations. In silico analysis revealed the greatest enrichment of A3G/F hotspot motifs occurs within the viral genomic sequences that encode for the most immunogenic CTL epitopes. Our ex vivo data are consistent with the potential of A3G/F mutations to promote CTL escape. Our observation that HIV has evolved strategically-placed A3G/F mutable hotspots in its genome to maximally direct A3G/F activity towards CTL escape is further evidence that A3G/F mutations likely impact adaptive immunity in vivo.In this study, we simulated A3G/F mutations of CTL epitopes of HIV and measured their effect on recognition by circulating CD8http://www.hiv.lanl.gov/content/immunology/tables/ctl_summary.html). A3G/F mutations on the 5′ dG located in A3G/F hotspots in the CTL epitope-coding genomic DNA were simulated in silico to generate mutated epitopes. For epitopes with multiple hotspots or those with potential for the generation of novel hotspots as a result of first-round mutation, multiple mutated versions were considered. Wild type (wt) and mutant (mut) forms of all selected epitopes were synthesized to ∼90% purity  and used to measure peptide-specific CTL responses from HLA-matched HIV+ individuals.The 9229 nucleotide-long genome of HIV isolate BRU was retrieved from the National Center for Biotechnology Information (NCBI) website. Isolate BRU was selected due to its representation of clade B, which is most prevalent in western countries + T cells. The ELISPOT assay to measure the frequency of HIV-specific CD8+ T cells has been previously described 6 PBMC. Even though the background frequency of IFN-producing unstimulated PBMC was negligible , we only considered >50 SFC/106 PBMC as a positive response. . Differences of >25% in SFC between wt and mut were considered significant.HIV-infected study participants were recruited through the Newfoundland and Labrador Provincial HIV Clinic in St. John’s, Newfoundland, and selected on the basis of their HLA class I types determined for previous immunological studies vs. outside CTL epitopes in each HIV gene was determined by scanning the BRU sequence. To normalize for gene size, the frequencies of hotspot occurrence were divided by the total number of nucleotides. These frequencies were calculated both for all A3G/F hotspots, and also solely for the most frequently targeted A3G hotspot (GGG). Subsequently, the ratio (R) of normalized hotspot frequencies inside to outside epitopes was calculated for each gene. In order to evaluate differences in R amongst various genes, an index was generated by determining the average of the R of all genes and dividing the R of each gene by the average. Thus, an index of >1 indicates an above average ratio of A3G/F hotspots inside to outside CTL epitopes. We further conducted the same type of analysis within each gene in order to compare hotspot frequencies between epitopes restricted to different HLA alleles .The number of hotspots  inside vs. mut epitopes were used to calculate statistical significance using Graphpad Prism 5.0.The wilcoxon signed-rank test was used to evaluate the statistical likelihood that introduction of APOBEC-related mutations into HIV CTL epitopes reduces recognition of those epitopes. Paired values for all Elispot comparisons of wt http://www.hiv.lanl.gov/content/immunology/tables/ctl_summary.html). This analysis for Gag is shown in http://www.allelefrequencies.net). We examined CTL epitopes in Gag, Pol and Nef because these contain the HIV peptides most prominently recognized by CD8+ T cells in silico. Epitopes for which the only possible mutation led to a stop codon or non-conservative changes in HLA anchor residues (thus impeding presentation by MHC class I) were excluded. Using these criteria, we selected and synthesized 27 wild type (wt) epitopes and 64 derivative mutants (mut) thereof (http://www.hiv.lanl.gov/content/sequence/QUICK_ALIGN/QuickAlign.html). We found that nearly all mut epitopes occur in the population. The selected wt and mut epitopes thus considered  tested against TSTLQEQIGW and 1 mut, P197 alone had a positive response that was 3-fold higher against the wt peptide (280 vs. 85 SFC/106 PBMC) (6 PBMC), whilst none had responses against the mut  (+ T cell recognition.We compared the CD806 PBMC) . Three o the mut . Eight s06 PBMC) . Respons06 PBMC) . Taken tvs. 910 SFC/106 PBMC)   . Five had positive responses exclusively against the wt peptide . Recogni06 PBMC) . Recogni06 PBMC) . Recogni06 PBMC) . LVGPTPV06 PBMC) . ALVEICT06 PBMC) . Altoget+ subjects were tested for CD8+ T cell responses against LSEGATPQDL and 3 muts. Five responded against the wt peptide  and 2 had lesser responses against the G to R mut   . Five su wt only . Overallex vivo reduction of HIV-specific CD8+ T cell responses by A3G/F mutations introduced into common HLA-A2, B44 and B57-restricted epitopes suggests that it would be advantageous for HIV to adapt towards maximizing A3G/F hotspots in genomic sequences encoding highly immunogenic or broadly presented CTL epitopes. To test this, we calculated the ratio (R) of A3G/F hotspot frequency inside vs. outside CTL epitopes within each HIV gene, normalizing for sequence length. We considered the frequencies of all A3G/F hotspots, but because A3G mutates HIV several fold more potently than A3F, and CCC (GGG on the plus DNA strand) is its preferentially targeted motif vs. outside CTL epitopes for each gene is reflected by an index that constitutes the ratio of the R value of each gene to the average R of all genes. Considering the index values for all hotspots, as well as GGG alone, Gag, Pol and Nef exhibited the highest indices  (The ctively) .vs. outside epitopes restricted to each HLA allele  CTL response. To examine this, we categorized epitopes on the basis of their restricting HLA . For each gene, we tallied hotspots inside A allele . We obseIn order to test if there is broad A3G/F hotspot enrichment within CTL epitopes, we compared the R of A3G/F hotspots inside to outside CTL epitopes of each gene to the R of hotspots inside to outside antibody epitopes in Env, which we considered as a control. The R for antibody epitopes in Env is 0.71 and 0.9 for all A3G/F hotspots or GGG, respectively. These are 1.3- to 3-fold lower than the R values for CTL epitopes of Pol (1.06 and 1.63), Gag (1.16 and 2.60) and Nef (0.89 and 2.45), but interestingly, up to 7-fold higher than the R values found in the CTL motifs of other HIV genes . The average R value of GGG inside to outside CTL epitopes in all genes is 2-fold higher than its counterpart for Ab epitopes across all genes . Collect+ CTL plays a key role in controlling viremia as illustrated by the correlation between disease progression and class I HLA genotype in vitro by producing truncated peptides that are more effectively processed and presented + T cell response in HIV+ cohorts to epitope variants adapted at the population level found that >50% of mut epitopes including hypothetical variants  retained their ability to stimulate CD8+ T cells The anti-HIV response by CD8+ T cell recognition of HIV. First, it is possible that some in silico-simulated A3G/F mutations may not have arisen in the subjects we tested. This is unlikely since we verified that all of our mut epitopes have been reported at the population level in a database of HIV CTL epitope variants (http://www.hiv.lanl.gov/content/sequence/QUICK_ALIGN/QuickAlign.html). Even if the occurrence of the mutation elicited a response in as low as ∼1 in 10,000 CD8+ T cells, it should be detectable by ELISPOT. It is possible that the decrease in CTL response to mut epitopes was a product of our particular experimental procedures. Our finding that several mut epitopes within Pol were equally well recognized Click here for additional data file.Figure S2A: Pol, B: Rev, C: Vif, D: Nef, E: Vpr, F: Env, G: Gag, H: Tat). The brackets indicate epitopes on the peptide sequence. Epitopes restricted to different HLA alleles are shown in different colors and the restricting HLA is indicated above the bracket. Broken lines display epitopes with no A3G/F hotspots and continuous lines show epitopes that harbor A3G/F hotspots.Map of all HIV-1 Bru isolate CTL epitopes across the viral proteome. HIV proteins are shown individually ((PDF)Click here for additional data file.Figure S3A: Pol, B: Rev, C: Vif, D: Nef, E: Vpr, F: Env, G: Gag, H: Tat). Colors differentiate sequences encoding CTL epitopes from the surrounding DNA. Colors do not otherwise correspond to any common features amongst genes or epitopes.Map of the viral genomic sequences that encode CTL epitopes in the HIV-1 Bru isolate plus-sense coding DNA. Sequence of each gene is shown ((PDF)Click here for additional data file.Table S1Relative enrichment of A3G/F hotspot inside/outside CTL epitopes in HIV genes. For each HIV gene, the frequency of A3G/F hotspots  was calculated in genomic sequences that encode for CTL epitopes and those that do not encode for CTL epitopes. Normalized frequencies were calculated by taking into account the total nucleotide length of the sequence. For each HIV gene, the normalized frequency of A3G/F hotspots inside sequences that encode CTL motifs was divided by the normalized frequency of A3G/F hotspots in sequences that do not encode CTL epitopes to generate a Ratio . The average of R-values was determined and each R-value was divided by the average to obtain the index value and allow for determination of the relative enrichment of hotspots within the CTL epitopes of each gene, as compared to the average ratio of hotspots inside to outside CTL epitopes. An index value >1 indicates relative enrichment.(PDF)Click here for additional data file.


Diploplastron affine is known as a common species of the rumen fauna in cattle and sheep. This protozoon is able to digest cellulose, whereas its amylolytic activity is not well known. The objective of the reported studies was to examine the ability of D. affine to digest starch and to use this polysaccharide to cover the requirement for energy. The enzymatic studies showed that the protozoal cell extract degraded starch to reducing products with the rate being equivalent to 2.4 ± 0.47 μmol/L glucose per mg protein per min. Maltose, maltotriose and a small quantity of glucose were the end products of starch degradation. The degradation rate of maltose was only 0.05 μmol/L glucose per mg protein per min. Two peaks in α-amylase and a single peak in maltase activity were found following molecular filtration of ciliate cell extract, whereas three starch-degrading enzymes were identified by a zymographic technique. Incubation of the bacteria-free ciliates with starch in the presence of antibiotics resulted in a release of volatile fatty acids with the net rate of 25 pmol per protozoan per h. Acetic acid followed by butyric acid was the main product of starch fermentation. The results confirmed the ability of D. affine to utilize starch in energy-yielding processes.The ciliate  Diploplastron affine is a common species of rumen fauna in the domesticated ruminants (Dogiel D. affine to digest starch and to use this polysaccharide to cover the requirement for energy.The ciliate s Dogiel . To dates Dogiel  whereas The ciliates were isolated from the rumen fluid of sheep. The cell extract for enzymatic studies was obtained by homogenization of purified protozoa and removal of the particulate matter by centrifugation. To restrict the bacteria, the ciliates were incubated overnight with ampicillin, streptomycin and chloramphenicol prior to the homogenization. The amylolytic activity of the ciliates was examined by quantification of products released during incubation of the cell extract with appropriate substrates. Fractionation of the crude enzyme preparation was performed by molecular filtration using a glass column packed with Sephadex G-150. Enzymes were identified by a zymography technique following separation of protein on polyacrylamide gel (Gabriel and Wang D. affine possesses enzymes degrading starch and its derivatives. Protozoal cell extract prepared from ciliates incubated with antibiotics released reducing sugars from starch, dextrin, maltose and isomaltose (Fig. Eudiplodinium maggii was studied (Bełżecki et al. D. affine ingested and fermented starch, releasing about 45 pmol VFA per protozoan per h. The control cultures produced only about 20 pmol VFA per protozoan per h (Table E. maggii was fractionated by ion exchange chromatography. On the other hand, three starch-degrading enzymes were identified by a zymographic technique following the separation of D. affine protein by native polyacrylamide gel electrophoresis (Fig. E. maggii crude enzyme preparation (Bełżecki et al. D. affine belongs to the rumen microorganisms which are able to digest starch and to utilize the obtained products as a carbon source in energy-yielding processes.Ciliate ose Fig. , Table 1 h Table  found onsis Fig. . For com


DFROBP showed low fat and high dietary fiber contents. The soluble and insoluble dietary fiber fractions were balanced, which is of importance for the health beneficial effects of fiber sources. DFROBP-containing muffins showed the same rapidly digestible starch content as the reference muffin, whilst the slowly digestible starch level increased with the addition of DFROBP. However, the resistant starch content decreased when DFROBP increased in the muffin. The addition of DFROBP to muffin decreased the predicted glyceamic index, but no difference was found between the muffins prepared with the two DFROBP levels. The sensory score did not show difference between control muffin and that added with 10% of DFROBP. The addition of DFROBP to bakery products can be an alternative for people requiring low glyceamic response.Orange is a tropical fruit used in the juice industry, yielding important quantities of by products. The objective of this work was to obtain a dietary fiber-rich orange bagasse product (DFROBP), evaluate its chemical composition and its use in the preparation of a bakery product (muffin). Muffins containing two different levels of DFROBP were studied regarding chemical composition,  Among current dietary trends, consumption of food products with reduced content of digestible carbohydrates has gained in popularity. Dietary fiber (DF) is a food ingredient that is neither digestible nor absorbed in the small intestine of the human. The development of new products with substantial DF contents is a strategic area for the food industry. Consumers are demanding foods that show two main properties: the first one refers to the traditional nutritional aspects of the food, whereas, as a second feature, additional health benefits are expected from its regular ingestion. Foods complying with these requisites are often called functional or nutraceutical foods.In a rapidly changing world, with altered food habits and stressful life styles, it is more and more recognized that a healthy digestive system is essential for the overall quality of life . DF playMost fractions  of DF are the major constituents of plant cell walls . Althougi.e., low in fat and digestible carbohydrates, high in fiber and low calorie content [™ by Natural Citrus Products  that is used in bakery products.Among good sources of fiber cereal grains, legumes, fruits , vegetables, nuts and seeds are of importance. These sources include citrus, as it exhibits 25–70% fiber content . Approxi content . Thus, o content ,10 and a content ; howeverFibers traditionally used for food processing are derived from cereals. Diverse studies have been conducted to obtain and assess the composition of dietary fiber-rich products obtained from by-products of diverse vegetable sources such as passion fruit, apples, pears, oranges, peaches, artichokes, asparagus, lemon, black currant, pear, cherry and carrot –14.Starch is the major digestible carbohydrate in human diet , represein vitro and in vivo by the macro-food properties , the presence of other dietary compounds as fiber and lipids, as well as anti-nutrients . Also the structure and physicochemical properties of native (raw) starch granules  may influence the kinetics and extent of the polymer digestion [The rate at which starch and other carbohydrates are digested and absorbed in the small intestine, has received great interest because of its association with the glycemic response and postprandial metabolism. Most baked goods contain free sugars and gelatinized starch, which have a readily dispersible in the food matrix. Starch digestibility can be affected igestion .It has been observed that co-ingestion of starch and soluble fibers results in slowered gastric emptying, which may also contribute to reduced postprandial blood glucose and insulin levels and thus influence satiety .Since and increased intake of DF is generally believed to be an effective way for prevention of chronic diseases, this ingredient is used in a variety of foods such as bars, cookies, soups, beverages, confectionery, snacks, in which has. Over the years, various fibers sources have fallen into and out of acceptance by the food industry and consumers alike. However, the use of fiber foods has continued to grow and expand, with ever-incrementing numbers of available applications .In view of the nutritional and technological relevance of dietary fiber, and the considerable volume of sweet orange bagasse disposal by the juice industry, the objective of this study was to obtain and characterize a dietary fiber-rich product from orange bagasse. The product was used to elaborate composite muffins whose chemical composition, starch digestibility, predicted glycemic index and sensory characteristics were assessed.2.2.1.Dietary fiber-rich orange bagasse product (DFROBP) exhibited low moisture content , similarIt is important to highlight the low total starch content of DFROBP (7.1 g/100 g dry sample) . Such a 2.2.Moisture content was similar in the two muffins containing DFROBP and lower than in the control product , showingSalvia hispanica L.) (2.25 g/100 g dry sample) or flaxseed (Linum usitatissimum L.) (1.41 g/100 g dry sample) [No differences between control and DFROBP containing samples were found regarding fat and protein contents . The low sample) .As in the case of DF, the total indigestible fraction (TIF) content of muffins increased with the addition of DFROBP . TIF lev2.3.The muffins elaborated with DFROBP had similar total starch contents . This obStudies in humans have demonstrated that incorporation of slowly digestible starch (SDS) and resistant starch (RS) in the diet can produce health benefits . Approxi® (1.56%), and different crackers (1.6–1.7%) [RS contents followed an inverse pattern to that observed for SDS. The values recorded in the DFROP-containing muffins were lower than those estimated in the reference product . This ma.6–1.7%)  are lowe.6–1.7%) .2.4.90) , particle size and viscosity, as well as by intrinsic characteristics such as structure and physicochemical properties of the starch substrate. In this sense, it is more meaningful to assess H90 as a physiologically closer parameter than the simple RDS/SDS content.Percentage of starch hydrolyzed at 90 min (H90)  and the Soluble components of dietary fiber  and 2 ca® (84) have been determined, while commercial cereal-based crackers had GI values ranging between 52 and 64 [in vivo GI of muffins prepared with DFROBP as this characteristic may be useful for the dietary management of people with impaired glucose tolerance.The pIG suggests important “slow digestion” features for the experimental muffins. A decrease of 10 points in pGI was recorded in muffins with DFROBP compared with the control muffin. Higher GI values for cornflakes (93) and Special K2 and 64 . It woul2.5.3.3.1.Citrus sinensis) from Ixthuatlán, Veracruz, Mexico were used. The peel of the orange was removed with an orange peeler and juice was extracted manually. The bagasse left was dried at 60 °C for 8 h in a tray dryer. The dry bagasse was ground in a manual mill  and sieved through a mesh number 40 (425 μm).Sweet oranges  were acquired in the local market. Wheat flour was provided by Selecta, S.A. de C.V. Mexico. The formulations of these muffins are shown in 3.3.et al. [Moisture content was determined by heating (110 °C for 3 h) using 2 g of sample. Ash, protein and fat were analyzed according to AACC methods 08-14, 42-11, and 32-25, respectively . Total set al. .3.4.The rapidly, slowly digestible and resistant starch fractions were determined with the procedure proposed by Englyst, Kingman & Cummings .3.5.in vitro rate of hydrolysis was measured using hog pancreatic α-amylase according to Holm et al. [The m et al.  with min90) values using the formula proposed by Goñi et al. [90) .The predicted glycemic index (pGI) was calculated from percentage of starch hydrolyzed at 90 min . Participants were untrained judges chosen at random from personnel at Centro de Desarrollo de Productos Bióticos, using a preference scale . A total3.7.Results were expressed as means of values ± standard error of the separate determinations. Comparison of means was performed by one-way analysis of variance (ANOVA) followed by Tukey’s test.4.A dietary fiber-rich orange bagasse product (DFROBP) with a total dietary fiber (TDF) content of 41.5% was prepared. The product increased the fiber content of experimental muffins by 40 and 63% compared to a control muffin. Although no difference in the rapidly digestible starch level was found between control muffin and those containing DFROBP, increased slowly digestible starch contents were recorded after the addition of this ingredient. However, resistant starch levels decreased with the addition of DFROBP. DFROBP-added muffins showed an importantly decreased predicted glycemic index, and the preference test indicated similar acceptability to the control and the 10% substituted muffin. Partial wheat flour substitution with DFROBP allowed production of prototype baked products containing high levels of TDF and indigestible fraction, features that may be of use in dietary regimes for people with different nutritional requirements.


Dear Friends,At the outset I extend my candid wishes to you and your family members for an eventful and prosperous New Year. Beginning January 1, 2011, I have taken over the responsibility of becoming the ‘Managing Editor’ for ‘Toxicology International,’ the official journal of the Society of Toxicology, India. I take this opportunity to express my gratitude to my predecessors for their vision and untiring efforts, which have given a strong base for setting the high standards that the journal continues to uphold, with significant visibility among scientists, academia, practitioners, and research students. I understand the challenges to compete with established journals in the area, in the present scenario, however; collective efforts of membership will help us reach the heights.Being a multidisciplinary forum for toxicology research, we welcome for publication, original research articles, with sufficient importance, novelty, and breadth of interest. In addition to that, articles presenting hypotheses and commentaries addressing current issues, which are of immediate interest to other investigators, are also welcome. Updates pertaining to areas of current importance and interests are also published in the form of mini-reviews. Besides clinical reports, occupational and safety evaluation, legal, risk, and hazard assessment, and impact of toxicants on humans, animals, and environment of sufficient novelty to warrant rapid publication, are also considered.http://www.journalonweb.com/ti/), which allows submission of articles with tracking of its progress till proof. It will be our endeavor to increase the number of issues in future. As you are aware that the journal is indexed at various sites, including ‘Pubmed’, it is high time to brain storm to have to procure an impact factor to improve the ranking of our journal among the journals published in the area of Toxicology.My goal is to continue with the policies developed by the earlier Editors, while striving to raise the standards, reputation, and visibility even further. In order to achieve this, the Advisory Board has been expanded, by including seasoned and active scholars working in diversified areas of toxicology. This will facilitate improvement in review processing for each submitted manuscript, by including one member, and eventually it would help in enhancing the consistency of better quality in the articles. I wish to develop a tradition of publishing special issues with a specific focal theme of high priority and relevance falling within the scope of the journal, which will further increase the journal’s strength, visibility, and circulation. A new feature, to provide a forum for the discussion and interpretation of data published in earlier issues of the journal, is also under consideration. We have achieved success in a difficult, but important task, of reducing the time between submission and publication through online electronic manuscript submission and a peer review system (I publically express my sincere thanks to all the members of the Editorial and Advisory Board, both past and present, the reviewers, authors, and all the dedicated personnel at Medknow Publications, in advance, for their contribution, cooperation, and services. Perhaps you may find some of my goals to be ambitious and far reaching, but I am confident that with your help and cooperation I will be able to achieve these targets.I look forward to continuing and building on the outstanding work of my predecessor, and the many other visionary and skilful people involved in creating and enriching this journal and guiding its development since the day of its inception. It is an appeal to forward suggestions to improve the quality of the journal, to deliver the best to the scientists and the society.Best wishes,


We examine the temporal evolution of digital communication activity relating to the American anti-capitalist movement Occupy Wall Street. Using a high-volume sample from the microblogging site Twitter, we investigate changes in Occupy participant engagement, interests, and social connectivity over a fifteen month period starting three months prior to the movement's first protest action. The results of this analysis indicate that, on Twitter, the Occupy movement tended to elicit participation from a set of highly interconnected users with pre-existing interests in domestic politics and foreign social movements. These users, while highly vocal in the months immediately following the birth of the movement, appear to have lost interest in Occupy related communication over the remainder of the study period. Information communications technologies play a crucial role in the development and persistence of many modern social movements The Twitter platform, like other information communication technologies, has the potential to confer a number of benefits to burgeoning social movements In related work To establish the extent of Occupy participant engagement with Twitter over time, here we study the total amount of Occupy-related traffic on the platform from September 2011 through September 2012. With respect to this measure of activity, we find that Occupy traffic has diminished by orders of magnitude relative to peak activity volumes in late 2011. This effect is evident even in concerted attempts to revive the movement's flagging levels of engagement, with activity returning to baseline within a week of May 1st, 2012 reoccupation efforts.Finding little evidence of sustained activity, we turn our attention to Occupy participants themselves, in hopes of understanding how these users were changed as a result of engaging with the movement online. Using a random sample of 25,000 Occupy users, we study changes in behavior at the individual level with respect to attention allocation and social connectivity. From this analysis we are left to conclude that, on Twitter, Occupy evoked interest from a highly-interconnected community of users with pre-existing interest in domestic politics and foreign social movements. Though we find statistically significant changes in political interests and social connectivity over the study period, the magnitude of these changes pales in comparison to the amount of attention these individuals allocated to the Occupy Wall Street cause.tweetshashtags. By following one or more accounts, a user creates a personalized feed that aggregates into a single, private stream the content produced by the followed accounts. Hashtags, short tokens prepended with a pound sign , allow the content produced by many individuals to be aggregated into a public, topic-specific stream including all the tweets containing a given token.Twitter is a social networking platform that allows individuals to consume content from and contribute content to streams comprised of 140-character messages known as Although by default each user's tweets are publicly visible, the audience for an individual's content is largely limited to his or her network of immediate followers, attaining greater levels of visibility only when it is rebroadcast by large numbers of other users. By including a hashtag in a tweet, however, an individual can contribute content to a high-profile stream, and thereby engage with users who might never otherwise see the content. It is this kind of communication, which represents engagement with a topically cohesive community of users unconstrained by social network structure, that is the primary focus of this study.retweet content produced by another individual, rebroadcasting it to his or her audience of followers, or mention another user in a tweet, which functions as a publicly-visible message targeting that individual.In addition to engaging with different content streams, users can interact with one another in two primary ways. A user can https://dev.twitter.com/docs/streaming-apis/streams/public). In addition to information about the content and users associated with a tweet, the Twitter streaming API provides timestamp metadata that allow for the historical reconstruction of the time series presented in this study.We rely on two primary datasets extracted over a 15-month period from an approximately 5–10% sample of the entire public Twitter stream , a second set relating to foreign social movements, and a third relating to domestic political communication. As we are interested exclusively in the attention allocation habits of Occupy users, we identified the set of hashtags relating to domestic political communication and foreign social movements by manually inspecting the 300 hashtags most frequently used by individuals in the random sample. All of the analyses in this article rely on time series describing changes to measured quantities over the course of the study period. Each time series is produced by computing a single statistic on disjoint sets of tweets partitioned into adjacent, temporally non-overlapping bins of At various times over the course of the study period, our system experienced service outages that affected our ability to collect data from the Twitter API. Amounting to 15 days in total, these periods are: September 29 to October 4, 2011; October 11–12, 2011; December 28–30, 2011; February 11–13, 2012; February 16–17, 2012; and May 28–31, 2012. Owing to the fact that the measures we employ reflect relative composition of the stream rather than its absolute volume, these outages do not unduly influence the statistical character of our results.Let us first focus to the total number of tweets in the Occupy corpus over the course of the year. In light of this finding, we wish to gain insights into the character of the individuals from which Occupy drew its support. We begin by studying how Occupy user interests changed in time, examining the frequency with which 25,000 random individuals produced content relating to one of three topics: Occupy Wall Street, foreign social movements, and domestic politics. Based on the random sample described in engaged user ratioAs we are interested in the behavior of individuals who were active on Twitter at a given time, we identify the set of users engaged user attention ratio. Among the set of users engaged with a topic, we next examine the extent to which that topic tends to dominate their content production activity. To accomplish this, let us consider, for each user As expected, a large fraction of users produced Occupy related content during the period of peak activity, with more than 40% of sampled users allocating on average 64% of their attention to the topic during the third week following the initial protests. However, this intense focus on the subject is not sustained over the course of the following year, with the engaged user ratio decaying to less than 5% in the last three months of the study period. Moreover, comparing the engaged user attention ratios from the first half of the period following the initial Occupy protests (With respect to foreign social movements and domestic political communication, we observe that users who would go on to engage with the Occupy movement online tended to exhibit interest in these topics before the initial protest activity in September, 2011. Comparing the engaged user ratios in the first 12 weeks of the study period with those observed during the last 12 weeks of the study period, we find a significant but small increase in domestic political communication activity. This conclusion is based on a two location t-test for a difference in sample means, in which we reject the null hypothesis (Finally, let us examine the extent to which Occupy users tended to interact with one another over the course of the study period. To this end we focus on the proportion of retweets and mentions produced by active users in the random sample that involved another user who produced at least one Occupy-related tweet during the year following the movement's inception. This proportion is computed with respect to all of a user's retweets and mentions, regardless of content, rather than just those related to Occupy Wall Street. Inspecting the 95% confidence interval bands in While interest and activity relating to the Occupy movement has substantially diminished, one could envision that increased levels of engagement with the political process online might constitute a positive outcome for the movement's participants. Along these lines, however, Occupy users remain barely changed, exhibiting a slight increase in attention paid to domestic politics and a slight decrease in attention paid to foreign social movements. Relative to the dramatic behavioral changes these users exhibited in the early stages of the movement, and the magnitude of Occupy-related communication in general, these changes constitute a somewhat underwhelming long-term effect.Similarly, a supporter of the movement might take as a promising outcome increased levels of interaction among Occupy users. Such a scenario could indicate that these individuals formed a more tight-knit community over the course of the year, creating social and communication bonds that may help to facilitate the efficient spread of information, potentially even reinforcing individual propensity for offline activity Taken together, these data suggest that, on Twitter, the Occupy movement tended to elicit participation from a set of highly interconnected users with pre-existing interests in domestic politics and foreign social movements. These same users, while highly vocal in the months immediately following the movement's birth, appear to have lost interest in Occupy-related communication over the remainder of the study period, and have exhibited only marginal changes in their attention allocation habits and social connectivity as a result of their participation.These findings should not be taken to suggest that the Occupy movement itself has failed, as an argument can be made that the movement played a role in increasing the prominence of social and economic inequality in the public discourse. Though it would be unreasonable to argue that users could have maintained the frenetic pace of Occupy's earliest days, it is doubtless that supporters may have hoped for a more sustained discourse than is evident from the near-complete abandonment of these once high-profile communication channels.


According to the United Nations' World Tourism Organization, there were an estimated 880 million international travelers in 2009, or roughly 13% of the world's population [National and international tobacco regulations (such as article 13 of the FCTC) have been promoted to prevent the tobacco industry from communicating with youth and other susceptible populations [Presented in vivid, colorful displays portraying brand names, themes, and logos Figure , and oftDuty free sales undermine the purpose of taxation and harms public health by encouraging personal consumption. Research from a number of countries has indicated that the percentage of smokers consuming tobacco from a duty free source ranges from below 1% in the US, Canada and Australia, and up to 3.8% and 5.6% in New Zealand and the UK, respectively -9. When The time has come to discuss the rational behind exemptions for airport advertising and duty free tobacco sales. Advertising and purchases within international travel should be governed by universal regulations such as those promoted by the FCTC and action should be taken towards their implementation at a global level as the tobacco industry does not recognize borders or boundaries ,10.Tobacco Induced Diseasesembraces the need for the collection of basic research on tobacco promotion and advertising within airports as a means for promoting evidence based advocacy.The authors declare that they have no competing interests.


The increase of aesthetic demands, together with the successful outcome of current implants, has renewed interest in the search for new materials with enough mechanical properties and better aesthetic qualities than the materials customarily used in implanto-prosthetic rehabilitation. Among these materials, zirconium has been used in different types of implants, including prosthetic abutments. The aim of the present review is to analyse current scientific evidence supporting the use of this material for the above mentioned purposes.We carried out the review of the literature published in the last ten years (2000 through 2010) of in vitro trials of dynamic and static loading of zirconium abutments found in the databases of Medline and Cochrane using the key words zirconium abutment, fracture resistance, fracture strength, cyclic loading.Although we have found a wide variability of values among the different studies, abutments show favourable clinical behaviour for the rehabilitation of single implants in the anterior area. Such variability may be explained by the difficulty to simulate daily mastication under in vitro conditions. The clinical evidence, as found in our study, does not recommend the use of implanto-prosthetic zirconium abutments in the molar area. Key words: Zirconium abutment, zirconium implant abutment, zirconia abutment, fracture resistance, fracturestrength, cyclic loading. Implantology has been one of the greatest odontological achievements in the last century. Once the stage when osteointegration was the only key to success has been overcome, the present challenge is to ensure both the functionality and the aesthetic value of the implant-supported prosthesis. The advantages of using ceramic components in implantoprosthetic rehabilitation can be summarized in the possibility to individualize the components, to take full advantage of their light transmission properties, excellent aesthetic qualities with colours resembling those of teeth and their great compatibility with preimplant soft tissues. Nevertheless, there are also disadvantages such as their lesser resistance to fatigue . Nowadays most companies in the sector offer ceramic abutments which may be prefabricated or manufactured and can also be prepared in a dental laboratory either by a technician following the traditional method or with the assistance of computer-based designs. The materials chosen for that purpose are: highly pure alumina Al2O3), TZP ceramics  or PSZ ceramics . TZP ceramics show superior properties to alumina due to such microstructural differences as density, size of particles  and their polymorphic mechanism against flaw propagation. The main reason for such superiority is the yttrium stabilization of the zirconia structure . These oxides can stabilize a relatively weak cubic structure below room temperature. On the other hand, if we add enough amount of stabilizing oxide, and the material is correctly processed, zirconium particles may be stored in a metastable tetragonal form at room temperature. All these materials are known as partially stabilized zirconium ceramics (PSZ). The transformation from the tetragonal to the monoclinic structure may serve to improve the mechanical resistance and fracture tenacity of PSZ ceramics. Used under stress in the area of a flaw, metastable tetragonal particles pass to a stable monoclinic structure, change which is known as martensitic transformation. The extension of the resulting volume is distributed around these particles close to the flaw thus compressing it and delaying its propagation until the stress is increased. This physical phenomenon ensures the reliability of this material for the production of implant-supported abutments . Zirconium oxide, or zirconia, shows similar mechanical properties to metals and its colour resembles that of teeth. Partially yttrium-stabilized zirconia shows the best properties for the purposes under discussion. Zirconia was first used with medical pur-poses in 1969. Helmer and Driskell introduced it as a new material for orthopaedic uses in the Symposium on Use of Ceramics as Surgical Implants. They presented it as a substitute for titanium or alumnia prostheses in hip replacement. The first zirconia abutment, Zirabut®  was produced in 1997, although many more were added later .The aim of the present review is to confirm scientific evidence in favour of the resistance of partially yttrium-stabilized zirconia abutments, commonly known simply as zirconia abutments, using dynamic and static loading trials. We carried out the review of the literature published during the last ten years (2000-2010) of in vitro trials of dynamic and static loading of zirconium abutments. In order to do so, we used the databases of Medline (Pubmed) and Cochrane. The key words used in the search were: zirconium abutment, zirconium implant abutment, zirconia abutment, fracture resistance, fracture strength, cyclic loading. First of all, we searched zirconium abutment OR zirconium implant abutment OR zirconia abutment and then we looked for fracture resistance OR fracture strength OR cyclic loading. Finally, we combined the keywords of both searches with the operator AND. In order to complete the search, we carried out a manual examination in the following publications of the articles available on the subject published in the last ten years: Journal of Prothetic Dentistry, Journal of Oral Rehabilitation, Quintessence International, Journal of Prosthodontics, International Journal of Oral Maxillofacial Implants, International Journal of Prosthodontics.We examined all the studies including in vitro fracture strength at a constant speed and dynamic loading trials of zirconia abutments used as implantoprosthetic abutment . We considered both studies in which the implantoprosthetic system was completely rehabilitated and those in which it was not. We found 9 articles whose main characteristics are shown in .In one of the trials on zirconia abutments  (WohlwenLikewise, Butz and cols.  in theirAramouni and cols.  in a recGehrke and cols.  in a repAdatia and cols.  publisheKerstein and Radke  comparedIn both instances, the fracture was due to small defects in their surface resulting from the fabrication process. AAZ® abutments had statistically lower probability of fracture at loads which resembled human mastication. However, both types of abutments failed at loads heavier than those occurring in daily mastication. Kim and cols.  studied An in vitro study carried out by Mitsias and cols.  comparedNorthdurft and cols.  introducThe literature so far published on this subject allows us to observe the behaviour of zirconium abutments under loads and pressure resembling daily mastication. We commonly find a wide variability of fracture force values among similar studies. The difficulty to simulate daily mastication resides in the great amount of factors involved in such process which complicate the design of this kind of tests. As a result, although the occlusal forces involved in mastication are well documented, there is no agreement on mean values. Leaving aside individual anatomic and physiologic characteristics, maximum occlusal force is reported to be located in the first molar region, with values ranging from 180 to 850 N, decreasing to values between 95 and 250 N in the incisive region -16. ConsAs regards fatigue trials, a mean load of 50 N during 2.4x105 cycles is considered to represent a year long of mastication. Such data are supported by the agreement observed between clinical and laboratory studies. In this sense, zirconium abutments still show some shortcomings, but we must bear in mind that the lesser the tension on repaired abutments the better their fatigue behaviour and the more satisfactory the results of in vitro studies on abutments without prosthetic crowns.Current clinical tests show the favourable outcome of zirconium abutments in the incisive region, even in the canine-premolar region, in instances of unitary implant-supported rehabilitations -19. Neve


Mammalian gene regulation is often mediated by distal enhancer elements. Despite this, mechanistic investigations of correlated expression have thus far been focused on proximal promoter regions, mainly because distal enhancers are not known for a vast majority of genes. However, distal enhancers of co-regulated genes are likely to have correlated activity.cis) and from different chromosomes, after accounting for HS autocorrelation affecting cis-pairs. Using non-parametric tests and controlling for dependencies, highly correlated pairs are compared with background pairs for: enrichment of co-occurring binding motifs; for correlated gene expression across the 15 cell samples sourced for HS data; for shared gene function; for evidence of interactions between shared enhancer-binding transcription factors (TFs) and chromatin-modifying enzymes; and for Hi-C evidence of pair co-localization. The relationship between correlated enhancers now established, we conclude by scaling this pairs perspective to the building and validating of an enhancer network.Using P300-bound regions as putative enhancers  and usinWe find that correlated enhancers tend to share common TF-binding motifs, and that several chromatin modification enzymes preferentially interact with these TFs. We show that we can predict correlated enhancers with 73% accuracy based only on the presence of shared motifs for specific TFs. Also, genes near correlated enhancers have correlated expression and share common function. Correlated enhancers coincide with spatially proximal genomic regions assayed by Hi-C in two different cell types. Finally we construct an enhancer network based on shared motifs and correlated activity, and show its high overlap with biological processes and pathways.Overall, our analysis suggests that functionally linked genes may be co-regulated by distal enhancers whose activities are regulated by common sets of TFs and mediated by both 3D chromatin structure as well as chromatin modification enzymes. Our work represents the first investigation of enhancer networks based on correlated activity across multiple cell types.


Spastic paresis in cerebral palsy (CP) is characterized by increased joint stiffness that may be of neural origin, i.e. improper muscle activation caused by e.g. hyperreflexia or non-neural origin, i.e. altered tissue viscoelastic properties . Differentiation between these components is hard to achieve by common manual tests. We applied an assessment instrument to obtain quantitative measures of neural and non-neural contributions to ankle joint stiffness in CP.Twenty-three adolescents with CP and eleven healthy subjects were seated with their foot fixated to an electrically powered single axis footplate. Passive ramp-and-hold rotations were applied over full ankle range of motion (RoM) at low and high velocities. Subject specific tissue stiffness, viscosity and reflexive torque were estimated from ankle angle, torque and triceps surae EMG activity using a neuromuscular model.p = .002) and tissue stiffness 2.1 times larger (p = .018) compared to controls. High tissue stiffness was associated with reduced RoM (p < .001). Ratio between neural and non-neural contributors varied substantially within adolescents with CP. Significant associations of SPAT (spasticity test) score with both tissue stiffness and reflexive torque show agreement with clinical phenotype.In CP, triceps surae reflexive torque was on average 5.7 times larger (Using an instrumented and model based approach, increased joint stiffness in CP could be mainly attributed to higher reflexive torque compared to control subjects. Ratios between contributors varied substantially within adolescents with CP. Quantitative differentiation of neural and non-neural stiffness contributors in CP allows for assessment of individual patient characteristics and tailoring of therapy. Cerebral palsy (CP) comprises a variety of non-progressive upper motor neuron (UMN) lesions occurring in the developing fetal or infant brain. The resulting movement and posture disorders are generally characterized by loss of muscle strength, i.e. paresis, improper muscle activation by e.g. increased reflexes and loss of coordination by e.g. flexion synergies. In addition, changes of tissue viscoelastic properties may modulate the characteristics of the primary motor disorders ,2. SpastDe Vlugt et al.  developeThe objective of the present study was to quantify neural and non-neural contributors to ankle joint stiffness in patients with spastic CP and to assess its validity and reliability. A quantitative discrimination between the neural and non-neural components of joint stiffness in CP gives insight in pathophysiological mechanisms and may provide a strong instrument for development of tailored intervention strategies and their follow-up.Twenty-three adolescents with CP (mean age (SD) and range: 14.9 (2.4) y, 12–19 y, fifteen male) were recruited from the outpatient clinics of the Rijnlands Rehabilitation Centre and the Department of Rehabilitation Medicine of the Leiden University Medical Centre, Leiden, the Netherlands. Table Subjects were seated with their foot fixated onto an electrically powered single axis footplate . Subjects were seated with their hip and knee positioned at approximately 80° and 70° of flexion respectively Figure . The thiMeasurements were performed on the most affected ankle of each patient and at the right ankle in case of controls. Maximum plantar and dorsal flexion angles were assessed by a gradually increasing flexion torque from 0 to a maximum value of 15 Nm. RoM was defined as the difference between the maximum plantar and dorsal flexion angle and used as boundary for the subsequent RaH rotations. During the RaH rotations, the ankle was rotated at 4 different angular velocities  over the individually assessed RoM, starting in maximal plantar flexion. RaH rotations were started at random time instants. The hold phase lasted 4 seconds after which the ankle was moved back again to the neutral position. Time to cover a complete RaH rotation did not exceed 15 sec. Rest periods of about 30 sec were introduced between each RaH rotation to avoid hysteresis effects . All RaHTo distinguish between the neural and non-neural contributions to ankle joint stiffness, a nonlinear neuromuscular model of the ankle joint was used by which the ankle torque was predicted and matched to the measured ankle torque using EMG and ankle angle as input. The model included a Hill-type muscle model to describe the torque contribution from muscle activation induced by stretch reflexes. Hill-type models account for the effect of muscle length and lengthening velocity on muscle force. Passive torque from viscoelasticity  was modeled by exponential force-length and force-velocity functions. Tendon stiffness (series elastic element) was assumed to be infinitely stiff . The fuli the sample time and n the number of data points used for the parameter estimation. Tmeasured,i is the measured ankle reaction torque and Tel,imod the predicted ankle torque. Those rotations exhibiting a VAF score lower than the mean VAF over all rotations minus 2 times standard deviation were excluded from analysis.With Primary outcome parameters were RoM, tissue stiffness and viscosity and torque from triceps surae (TS) and tibialis anterior (TA) stretch reflexes. As passive tissue stiffness and viscosity strongly depend on joint angle, values at the maximal common dorsal flexion angle of all subjects were calculated for inter-subject analysis. This particular angle was chosen as exhibiting probably the largest contrast between subjects . Model sDifference in RoM between patients with CP and healthy controls was tested using an unpaired t-test. A linear mixed model was used to determine the difference in primary outcome variables between healthy controls and patients with CP (random factor) and to assess the effect of velocity . Stepwise linear regression procedures and one way ANOVA with Bonferroni correction were applied to assess associations of primary outcome variables with RoM and secondary outcome variables i.e. speed of ankle rotation, age, gender, GMFCS , AshwortOne subject  could not complete the RaH measurement due to insufficient relaxation and in this particular case only the RoM was used. In 2 other healthy subjects 1 RaH rotation had to be excluded due to technical problems. In total, 16 of the 256 RaH rotations from 9 subjects (8 CP), were excluded due to poor model fits  or insufficient relaxation . The VAF of the remaining RaH rotations was above 98.9%, meaning that the model could well describe ankle torque dynamics.t = 2.10, p = .044), see Figure RoM in dorsiflexion was significantly smaller in CP  and was significantly larger in CP compared to healthy controls , see Figure F = 9.86, df = 3, p < .001). We found no significant difference between the groups regarding ankle viscosity .Tissue stiffness was independent of velocity  and the difference increased with velocity . TS reflexive torque showed a large variation within the group with CP. TA reflexive torque was not significantly different between CP and healthy controls  and did not change with velocity .TS reflexive torque  was on average 4.2 times higher than reflexive torque at the highest ankle rotation speed (120 deg/sec) with a standard deviation of 3.3 indicating a substantially variation between subjects. Total explained variation of reflexive torque (120 deg/sec) by tissue stiffness (15 deg/sec) was 38%. Association between tissue stiffness and reflexive torque was low (ICC: less than .5).β = 15.8 se 4, p = .001). For reflexive torque, both SPAT score and age were significant positive contributors . Tissue stiffness , TS reflexive torque  and RoM (p = .001) differed significantly with respect to SPAT but not Ashworth score . It was suggested previously that the role of passive stiffness may increase over reflex activity with age in children with spastic diplegic CP  and thatCorrelation of SPAT score with both tissue stiffness and reflexive torque underlines the fact that it is difficult to split the neural and non-neural component with the manual tests such as SPAT. In future work we will measure more patients and with a wider range of GMFCS and SPAT scores to study this correlation more extensively.The present and former  studies TS reflexive torque was more dominant in CP than in stroke . For theTissue stiffness and reflexive torque could both be reliably estimated: tissue stiffness especially at the low ankle rotation speed and reflexive torque at the high speeds. This illustrates the feasibility of the method to distinguish contributors to joint stiffness on an individual basis. As expected, reflexive torque and not tissue stiffness was significantly influenced by ankle rotation speed and especially tissue stiffness was associated with RoM at low speed. Significant associations of SPAT score with both tissue stiffness and reflexive torque show agreement with clinical phenotype.We selected patients with CP with a relatively high GMFSC score (median of 1) to create a homogenous population. This limits validity of the present study and prevents more extensive elaboration of the relation with clinical phenotype which will be important for goal directed therapy. There were only 3 patients with a GMFCS score higher than 1, so correlations of GMFCS with neuromuscular parameters could not be studied well. The present study did not differentiate contributions of gastrocnemius and soleus and was performed at 1 knee angle. Joint stiffness should also be assessed in relation to functional movements, like walking using static and dynamic measurements . In thisFuture work will comprise the assessment of Achilles tendon stiffness by ultrasound measurements, the assessment of joint stiffness as a function of ankle rotation angle in a more detailed way, measurements at different knee angles and under functional (loaded) conditions.Using a novel instrumented assessment technique, patients with CP showed a smaller RoM and higher tissue stiffness and reflexive torque compared to control subjects. Good reliability and validity of the assessment technique combined with considerable intra-individual variance are a base for individual tailored therapy.The authors declare that they have no competing interests.KGG performed the analyses and wrote the analysis software, assisted in the experiments and wrote the manuscript. EV designed the experiment, wrote the processing software, assisted in data processing and interpretation and writing the manuscript. JG designed the experiment, assisted in data processing and interpretation and writing the manuscript. HHM designed the experiment and critically reviewed the manuscript. DW conducted the experiments and recruited the patients and critically reviewed the manuscript. RWH performed the clinical assessment and critically reviewed the manuscript. JA took part in discussions on the outcome and assisted in writing the manuscript. CM designed the experiment, performed the clinical assessment, assisted in data processing and interpretation and writing the manuscript. All authors read and approved the manuscript.


Loss of anterior tooth causes aesthetic and functional disharmony. Although no restorative material can approach the appearance of intact tooth enamel, glass ceramic, at the increased risk of brittle fracture, can mimic original tooth color better than the other restorative options. The newest zirconia material comes with unparalleled individualization in aesthetics and optimal physical properties. One of the basic principles of tooth preparation is conservation of tooth structure. This clinical report describes the replacement of maxillary and mandibular incisor with latest generation zirconia adhesive fixed partial denture. The authors have achieved unmatched aesthetics with newer high translucency zirconia. In the field of indirect restorative materials, it has always been desirable to have the right blend of aesthetics and durability. The zirconia material conquers all domains of dental restorations where extreme precision, strength, and aesthetics are required. The recently introduced high performance zirconia adds to its appearance by having improved translucency, that is, although inferior to glass ceramic but of course without as much as two thirds reduction in flexure strength.With respect to dental restoration, aesthetics means color that is matched perfectly in lightness to its adjacent teeth and translucency. Translucency refers to having the appearance between complete opacity and complete transparency. In order to produce lifelike restoration, the light must be transmitted and reflected as it does with the natural tooth and it's done.Lava plus high translucency zirconia is recently introduced by 3M ESPE . It is an after comer of original Lava zirconia, which is considered as one of the most successful ceramic systems. The newer version can be fabricated with hand layered, pressed, or monolithic options. This material is available with the widest range of dyeing liquids and Lava plus effect shades for excellent color match with VITA classical shade guide and unprecedented customization. It is claimed to have the only zirconia system with direct conversion to VITA system 3D-MASTER shades. The translucency of zirconia material is determined by impurities and structural defects as it causes light absorption and scattering, respectively. The high quality processing of Lava plus minimizes the effects of impurities and structural defects. The translucency of zirconia material is reduced by different refractory index and segregation nature of the alumina which is added into zirconia for aging stability. In Lava plus zirconia, the alumina content is reduced to 0.1 wt% and distribution is improved while maintaining the aging stability of the material .The strength of new zirconia material is no way inferior to Lava zirconia. The high strength of the material allows thinner walled restorations to add to the translucency and conserves the tooth structure, of course.The anterior restorations in the following case illustrate the use of this material to achieve highly aesthetic result.A 52-year-old man was diagnosed with partial anodontia with missing maxillary and mandibular right central incisor . VariousIn the next appointment, the provisionals were carefully removed. The Lava plus zirconia frameworks fabricated with CAD/CAM were evaluated intraorally for marginal fit, occlusion, and so forth. Subsequently, traditional hand layered technique was applied to allow restorations that exactly mimic the nature .μm aluminium oxide grain and treated with alloy primer. The prepared enamel surfaces were isolated, cleaned with prophylaxis paste, rinsed with water, air-dried, and etched with 37% phosphoric acid for 20 seconds. The dual cure adhesive resin cement  was used for luting the restorations to the abutment teeth. Excess cement was removed by hand waving technique, after brief tack light cure for 1 second, while holding the restoration in place. Restorations were light- cured according to manufacturer's recommendations for 20 seconds from all possible aspects (Figures The intaglio surface of the prostheses was grit blasted with <50  Figures . The pat2. Veneering of Lava plus zirconia restoration has three options as discussed above. The Lava plus monolithic veneering technique is selected when the clinical situation demands high strength or when there is reduced interocclusal space. The technique for bonding the restoration is essentially the same as with any other zirconia material. However, conventional cements can be used with traditional anterior FPD made of high translucency zirconia which is an added advantage over glass ceramics.High translucency zirconia is an option worth considering for restorations that need to be aesthetically superior and serve patient well for years. For Lava plus anterior adhesive bridge, the selected case should not have deep bite, heavy bruxism, and mobile abutments. The clinical protocol described in this article is straightforward and exactly the same as recommended. Considering laboratory protocol, the recommended connector cross section is 7 mmThis clinical report illustrates a conservative treatment with Lava plus zirconia adhesive FPD for the replacement of maxillary and mandibular incisor. The mechanism behind optimized translucency and color is described with case selection, specific tooth preparation steps, and clinical procedure involved in the newer high translucency zirconia adhesive FPD.


Sir,This is with reference to the comments on our article.2 RespectThe established drug efficacy studies/concepts need repetition in different gene pool/geographical area to verify the optimal results in diverse situations.4 The repApart from the efficacy of hydroxyurea, we also had our concern about safety aspects throughout the study period.Even the negative results are welcome in scientific community and some of the journals are fanatical for this job. We have conducted a scientifically sound, ethically approved, prospective study in sickle cell disease endemic tribal area which itself holds some logical merit to make a place in indexed journal.Indian Journal of Pharmacology has shown a full accountability and scientific conclusion to decide on this study in their much valued journal.Our main objective was to recommend a wider adoption of hydroxyurea for treatment in areas of high prevalence of sickle cell disease, which is still lacking. The


Situs inversus is a rare congenital anomaly characterized by the complete inversion of thoracic and abdominal organs. Liver transplantation in such patients or from donors in situs inversus is technically challenging because of the reversed anatomic structures. A small number of successful liver transplantation cases concerning situs inverus in either recipients or donors have been recently reported with different graft position and orientation. Here we reported an extremely rare case of liver retransplantation from an ABO incompatible situs inversus donor to an adult situs inversus recipient.A 53-year-old complete situs inversus man developed graft failure due to severe biliary complication after his first liver transplantation from a situs solitus donor. Re-transplantation was performed using a graft liver from a likewise situs inversus donor. Although the blood type between donor and recipient was incompatible, the post-operative outcome was excellent under proper prophylaxis to the antibody-mediated rejection.To the best of our knowledge, this is the first report of liver transplantation from situs inversus to situs inversus in adult recipient. Liver transplantation using situs matching donor makes the procedure much easier at the surgical point of view, which has a benefit of less potential surgical complications. Furthermore, ABO-incompatibility is acceptable for donor allocation in cases that both donor and recipient are situs inversus. Situs inversus (SI) is a rare congenital anomaly characterized by the complete inversion of thoracic and abdominal organs with an estimated incidence in humans about 0.025% to 0.005% live births . The exaHere we report a successful case of LTx from a SI donor to an adult SI recipient. Our case is particular rare and unique because: 1. The patient required re-transplantation due to graft failure after the first LTx from a normal donor; 2. The whole graft from the donor with SI was implanted; 3. The ABO blood type between donor and recipient was incompatible.A 53-year-old male was referred to our hospital in September 2012 for re-transplantation due to severe biliary complication after the first LTx. His blood type was “O”. Complete SI without other abnormalities was identified when he was a teenager. Following splenectomy and hepatectomy due to hemorrhage after hepatitis B related cirrhosis and hepatocellular carcinoma respectively, he underwent LTx from a deceased donor with situs solitus because of tumor recurrence in 2005 at the Eastern Hepatobiliary Surgery Hospital, Shanghai, China. Since half a year after LTx, he suffered episodes of biliary complication characterized as jaundice and fever. Bile duct stents were inserted several times. The total bilirubin fluctuated between 70-200 μmol/L and it was gradually elevated to about 400 μmol/L before he was listed for re-transplantation. He had severe skin pruritus and sporadic slight hepatic comma.The donor was a 32-year-old male who suffered brain injury in a motor vehicle accident and was pronounced dead shortly after admission. Image studies showed that he was situs reversus. His blood type was “B”. Given the organic situs match, the donor liver graft was then allocated to the recipient. No technical difficulties were encountered during the procurement. The liver graft showed completely inverted structure but without vessel anomaly after trimming the excess tissue on back table Figure .After careful laparotomy in recipient, a normal oriented liver was transplanted showing deeply bilious appearance with blunt right lobe and shrunken left lobe. The whole graft was severely rotated to the left upper quadrant. Excision of the original liver graft was performed with preservation of recipient inferior vena cava (IVC). The new liver was then brought onto the field as its original position. The structure of IVC, hepatic artery, portal vein and common bile duct of the graft were in line with the recipient’s corresponding structures. A piggy-back technique with some modifications was applied for cavo-cavostomy. In brief, a longitudinal midline incision on the posterior wall of donor suprahepatic IVC was cut to make a triangulated and wide orifice, which was end-to-side anastomosed to the corresponding similar incision in the anterior wall of recipient IVC from the orifice of hepatic veins. Donor infrahepatic IVC was ligated. The portal vein was end to end anastomosed. The donor’s common hepatic artery was anastomosed to the confluence of the recipient’s proper hepatic artery and gastroduodenal artery. Cholecystectomy was performed as usual and biliary reconstruction was achieved via end-to-end choledochocholedochostomy Figure A, B.To reduce the risk of complications from ABO mismatch, the patient was treated with the protocol of our center: 600 mg Rituximab  before transplantation and a total of 200 g intravenous immune globulin during operation and the first ten consecutive days post transplant. The maintenance immunosuppression included tacrolimus, mycophenolate mofetil and steroid. The post-operative outcome was uneventful. He was doing well with excellent graft function at 11 months follow-up. Computed tomography assessment demonstrated the graft in normal appearance in the up left quadrant Figure .Because of the rare occurrence of SI and the complexity of the situation, there remains limited experiences of LTx in patients with this problem . To dateIn fact, partial LTx from either deceased or living donor has been performed more because such approaches may minimize the difficulties with a smaller graft ,16. NeveThe present case was an ABO-incompatible LTx that was often performed in emergencies when ABO-compatible graft was unavailable. It was used to be a controversial issue because of the high risk of antibody-mediated rejection due to preformed anti-ABO antibodies. Various methods have been applied to improve the outcome of ABO incompatible LTx, including graft local infusion, use of rituximab, intravenous immune globulin, plasma exchange, and splenectomy ,18. EspeThe current case is the first report in adult LTx from SI donor to SI recipient. Form the surgical point of view, the same mirror-image liver vessel structure makes the present procedure much easier and thus have less potential surgical complications. Given the limited experiences of LTx in SI, if situs matches, we suggest that it’s better to allocate SI donor to SI recipient. ABO-incompatibility is acceptable for donor allocation in cases that both donor and recipient are SI.Written informed consents were obtained from both the recipient of the transplant and from the next-of-kin of the donor for publication of this case report and all accompanying images. The copies of the written consent are available for review by the Editor-in-Chief of this journal.SI: Situs inversus; LTx: Liver transplantation; IVC: Inferior vena cava; GB: Gallbladder; HA: Hepatic artery; PV: Portal vein; CBD: Common bile duct.The authors declare that they have no competing interests.YS reviewed the data and literature and wrote the manuscript. GH collected and reviewed the literature. ZW and YJ participated in the donor procurement. YS, ZM and SY participated in the operation and the treatment of recipient. ZS contributed to revising the manuscript. All authors have read and approved the final manuscript.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-230X/14/46/prepub


Arterial (aortic) stiffness is a well-recognized pathophysiological change that plays a significant role in the determination of risk factors for various cardiovascular diseases. MeasureIn-vivo aortic MRE and MRI was performed on 8 healthy volunteers Ages 18-35yrs). All imaging was performed using a 3T-MRI Scanner . The volunteers were positioned in the supine position and placed head first in the scanner. 60Hz mechanical waves were introduced in to the aorta using a pneumatic diver. A GRE-M5yrs. AllFigure This study demonstrated the feasibility of comparing MRE based stiffness estimates and PWV based stiffness estimates in the same imaging plane of the aorta.


Thus, the identification of new I-Ag7 and HLA-DQ8 epitopes would be of great help to further experimental and biomedical manipulation efforts. In this study, a novel GPS-MBA  software package was developed for the prediction of I-Ag7 and HLA-DQ8 epitopes. Using experimentally identified epitopes as the training data sets, a previously developed GPS (Group-based Prediction System) algorithm was adopted and improved. By extensive evaluation and comparison, the GPS-MBA performance was found to be much better than other tools of this type. With this powerful tool, we predicted a number of potentially new I-Ag7 and HLA-DQ8 epitopes. Furthermore, we designed a T1D epitope database (TEDB) for all of the experimentally identified and predicted T1D-associated epitopes. Taken together, this computational prediction result and analysis provides a starting point for further experimental considerations, and GPS-MBA is demonstrated to be a useful tool for generating starting information for experimentalists. The GPS-MBA is freely accessible for academic researchers at: http://mba.biocuckoo.org.As a severe chronic metabolic disease and autoimmune disorder, type 1 diabetes (T1D) affects millions of people world-wide. Recent advances in antigen-based immunotherapy have provided a great opportunity for further treating T1D with a high degree of selectivity. It is reported that MHC class II I-A In 2009, the first-in-human beings Phase I clinical study reported that proinsulin peptide injection is both well tolerated and safe Besides immunosuppressive drugs and antibody-based immunotherapies, antigen-based “tolerogenic” immunotherapy has attracted considerable attention as a third-generation approach, particularly for its highly selective targeting of aberrant T cells in silico prediction of MHC-binding epitopes has emerged as an efficient approach to generate useful information for the purposes of biomedical design http://mba.biocuckoo.org/ links.php). For example, the prediction results of SYFPEITHI g7 is the only expressed MHC class II molecule in the NOD mouse g7 or HLA-DQ8 epitopes et al. developed the first online server of PREDNOD for the prediction of I-Ag7, and the two MHC class I molecules Kd and Db binding peptides in the NOD mouse et al. used an expectation-maximization alignment algorithm to design computational programs for the prediction of I-Ag7g7 and HLA-DQ8, although they were developed for the comprehensive prediction of a variety of MHC class I and/or II binding peptides. Currently, although a number of computational studies have been performed, only MHC2Pred As a complement to labor-intensive and time-consuming experimental assays, the g7 and HLA-DQ8. The experimentally identified epitopes were obtained from the scientific literature, and a modified Gibbs sampling approach was adopted to determine the core nonamers in these epitopes. For training and prediction, a refined GPS algorithm g7 epitopes and vice versa, the results show that I-Ag7 and HLA-DQ8 recognize highly similar peptide profiles. With this powerful tool, we predicted potentially novel I-Ag7 and HLA-DQ8 binding peptides from T1D-associated epitopes, which bind to other types of MHC molecules. All of the experimentally identified T1D antigens together with their epitopes were absorbed into TEDB 1.0. The ab initio predicted epitopes were also provided. Taken together, the prediction and analysis results are helpful for further experimental investigation, and the GPS-MBA can serve as a practically useful adjunct program for experimentalists. The online service and local packages of GPS-MBA 1.0 were implemented in JAVA and freely accessible for academic research purposes at: http://mba.biocuckoo.org.In this work, we developed a novel GPS-MBA software package for the prediction of I-Ath, 2011) with the keywords “I-Ag7 peptide”, “HLA-DQ8 peptide”, or “Type 1 diabetes epitope”, we collected 318 experimentally verified and naturally processed mouse I-Ag7 binding peptides in 177 proteins, and 134 human HLA-DQ8 epitopes from 84 proteins (http://www.uniprot.org/uniprot/).A search of the scientific literature from PubMed  and negative (−) data sets for testing. The 318 I-Ag7 and 134 HLA-DQ8 epitopes of known length were regarded as the (+) set. If at least one predicted nonamer is fully located in the epitope region, the epitope is predicted as a positive hit. To avoid any bias, all of the 9-mer lengths of the same proteins which were either not covered or not fully covered by the original epitopes were taken to be the (−) set.In our data set, the length of most of the epitopes varies from 9∼30aa. Thus, we adopted a refined Gibbs sampling approach By further literature review, we collected 203 T1D-associated epitopes in 25 proteins which have the potential to be recognized by other types of MHC or unknown molecules .Sn), specificity (Sp), accuracy (Ac) and Mathew's Correlation Coefficient (MCC). The measurements were defined as below:As previously described n-fold) cross-validations were performed. In the LOO validation, each core nonamer in the data set was picked out in turn as an independent test sample, and all the remaining core nonamers were regarded as training data. This process was repeated until each nonamer was used as test data one time. In the n-fold cross-validation, all the (+) core nonamers and (−) nonamers were mixed and then divided equally into n parts, keeping the same distribution of (+) and (−) nonamers in each part. Then n-1 parts were merged into a training data set while the remnant part was taken as a testing data set. This process was repeated 20 times and the average performance of n-fold cross-validation was computed. Furthermore, the Receiver Operating Characteristic (ROC) curves were drawn, and AROC (area under ROC) values were calculated.To evaluate the prediction performance and robustness, the leave-one-out (LOO) validation and 4-, 6-, 8- and 10-fold  sites in proteins A and B as:The basic hypothesis behind the scoring strategy is that similarly short peptides would exhibit similar 3D structures and biochemical properties Score denotes the substitution score of the two amino acids of A[i] and B[i] in the BLOSUM62 at the position i. If S<0, we redefined it as S = 0. A given nonapeptide is then compared with each of the 9aa core peptides from the training data in a pairwise manner to calculate the similarity score. The average value of the substitution scores is regarded as the final score.Sn, Sp, Ac and MCC under different cut-off values. Thus, we fixed the Sp at 90% and compared the Sn values of the LOO validation.The performance improvement procedure is comprised of two steps, weight training (WT) and matrix mutation (MaM). To evaluate the degree of performance improvement, we calculated the scores for all sites of the training data set in each time. By gradually increasing the threshold, we computed the A and B was updated as follows:In this step, the substitution score between the two 9-mer peptides iw value is the weight of the position i. Again, if S′ is <0, we redefined it as S′ = 0. Then we randomly picked out the weight of any position for +1 or −1 and re-computed the LOO result. The manipulation was adopted if the Sn value was increased. This process was continued until the Sn value did not increase any further.Initially, the weight of each position was defined as 1. The Sn value by randomly picking out an element of the BLOSUM62 matrix for +1 or −1. The process was repeated until convergence was reached. Selecting a different initial matrix, e.g., BLOSUM45 will generate a convergent result if the training time is sufficient (Data not shown).The aim of this step is to generate an optimal or near-optimal scoring matrix from an initial matrix. We re-calculated the LOO result to improve the The online service and local packages of GPS-MBA 1.0 were implemented in JAVA. For the online service, we tested GPS-MBA 1.0 on a variety of internet browsers, including Internet Explorer 6.0, 8.0 and 9.0, Mozilla Firefox 8.0 and Google Chrome under the Windows XP and Windows 7 Operating System (OS), Mozilla Firefox 8.0 and Google Chrome under Fedora Core 15 and Ubuntu 10.04 LTS (Linux), and Safari 5.1.1 under Apple Mac OS ×10.5 (Leopard) and 10.7 (Lion). For the Windows and Linux systems, the latest version of Java Runtime Environment (JRE) package (Java SE 5.0 or later versions) should be pre-installed. However, for Mac OS, GPS-MBA 1.0 can be directly used without any additional packages. For convenience, we also developed local packages of GPS-MBA 1.0 which support the three major Operating Systems Windows, Linux and Mac OS X.g7 epitopes and 134 HLA-DQ8 binding peptides of various lengths from 8∼30aa  is the average final similarity score compared with the other nonamers, while S is the similarity score between Pi and Pj. The 9-mer with the maximal Score(Pi) is sampled and then updated to the new nonapeptide Pi , WT and MaM The series of GPS algorithms contain two computational procedures of a scoring strategy and performance improvement k-means clustering approach can be used to classify positive data into multiple groups g7 and HLA-DQ8 recognition When the training data set is large, the 174ERQNGVLNSWTDQDS188, identical to 46–60 in the Igκ chain C region) was sequenced as an I-Ag7 epitope by MS/MS analysis g7 epitopes of 4APTVSIFPP12, 49NGVLNSWTD57, 68SSTLTLTKD76, and 69STLTLTKDE77 were predicted , while the other three predicted hits are available for further experimental investigation.Taken together, the GPS algorithm has been improved, and the performance improvement process only required the two steps of WT and MaM. The training order was shuffled for better performance, and the online service and software packages of GPS-MBA 1.0 were implemented in JAVA. As an example, the prediction results for the mouse Igκ chain C region (UniProt ID: P01837) are shown . Previouredicted . The 49Ng7  and HLA-DQ8 (0.1534), respectively  was still much better than MHC2Pred (47.80%) and RANKPEP (64.15%)  . Once ag15%) (g7  and HLA-15%)   . Also, t (33.5%) . In partectively . Howeverpeptides . Also, tpeptides . In thispeptides . Taken tg7 or HLA-DQ8 epitopes in 245 antigens  provides an interface for querying the TEDB database with one or several keywords such as TEDB ID, MHC Type, or UniProt Accession, etc  or juvenile diabetes, is a chronic metabolic disorder and autoimmune disease that takes a heavy toll world-wide g7 or HLA-DQ8 epitopes, the online services have been not available over the internet g7 and HLA-DQ8, they were actually designed for the general prediction of MHC class I and/or II epitopes g7 and/or HLA-DQ8. In this study, we focused on the prediction of I-Ag7 and HLA-DQ8 by constructing the GPS-MBA software package. By comparison, the performance of GPS-MBA is much better than MHC2Pred and RANKPEP  and HLA-DQ8 (24/134). Again, in the non-redundant core nonamers, only 22 9-mer peptides were found to bind with both I-Ag7 and HLA-DQ8. Based on these results, we propose that the sequence profile of I-Ag7 is intrinsically similar to that of HLA-DQ8.Here, we used the WebLogo server g7  and HLA-g7 , respectg7 . P4 is wnonamers  and not nonamers . Althougprofiles . We specab initio predictions with GPS-MBA were also integrated into TEDB. Thus, such an integrative platform should prove to be useful for experimentalists. We believe that computational analysis, together with subsequent experimental identification, will help advance the study of T1D into a new and highly productive phase.In addition to the predictions of GPS-MBA, all of the experimentally validated T1D-associated epitopes were collected in the online database of TEDB 1.0. The Table S1g7 epitopes.The core nonamers of the mouse I-Aa. Epitope, the original epitopes; b. Position, the positions of core nonamers in the protein sequences; c. Core nonamer, the finally aligned core nonamers were marked in red; d. The protein sequence was retrieved from the UniParc (UniProt Archive) Database (http://www.uniprot.org/help/uniparc).(XLS)Click here for additional data file.Table S2The core nonamers of the human HLA-DQ8 epitopes.(XLS)Click here for additional data file.Table S3From the scientific literature, we collected 203 epitopes in 25 proteins, with 70 MHC class I epitopes, 84 MHC class II binding peptides, and 49 epitopes for which the MHC molecules are still unknown. The detailed prediction results of GPS-ARM were provided. a. UniProt, the UniProt accession numbers of T1D antigens; b. Pos., the position of the original known epitopes; c. Peptide, the experimentally identified epitopes; d. MHC Type, the experimentally identified MHC molecules that recognize the epitopes; e. Pre. Pos., the predicted position of the binding peptides; f. Pre. Peptide, the predicted core nonamers; g. Pre. Type, the predicted MHC molecules of I-Ag7 and HLA-DQ8 that potentially recognize the core 9-mers.(XLS)Click here for additional data file.


A highly respected, renowned academician of his generation, Dr. Saleh Zahedi-Asl passed away on the 3rd November 2013, leaving behind a lasting legacy. His sudden, totally unexpected demise left us all shocked and bewildered, but forced to come to terms with the tragic loss. Dr. Zahedi was the Editor-in-Chief of both the International Journal of Endocrinology and Metabolism and the Iranian Journal of Endocrinology and Metabolism, and the Deputy Director of the Research Institute for Endocrine Sciences (RIES), Shahid Beheshti University of Medical Sciences, Tehran. In the past he had been the editor in-chief of the Scientific Medical Journal of Ahvaz University of Medical Sciences (AUMS). A member of several editorial boards, both international and national, research councils and national examination boards and societies, Dr. Zahedi was never intimidated by controversy, did not always concur with current trends and was always available to and willing to corroborate with and counsel authors within and outside Iran.Born in Ardebil, Iran, on August 29th 1951, Dr. Zahedi graduated with an honors bachelor’s degree in basic sciences from the University of Tabriz (1973) and went on to obtain his master’s in physiology from the university of Jundishapur, Ahvaz (1978). Dr. Zahedi then proceeded to the University Of Newcastle Upon Tyne, UK for his Ph.D. (1981), during which he developed radioimmunoassay for insulin, two kinds of glucagon (pancreatic & gut) and GIP (glucose-dependent insulinotropic pepetide). He was granted a MRC postdoctoral fellowship to work with Dr. John Brown on projects involving raising monoclonal antibodies to hormones, extraction, purification and sequencing of new biologically active peptides. He had published over 108 papers in Persian and 81 in English and edited one book and translated four books from English to Farsi. Dr. Zahedi’s publications include many papers on both animal and human research, as well as some valuable papers in the field of endocrinology and gastro-intestinal physiology. He had recently worked on the developmental effects of thyroid hormones and demonstrated for the first time that maternal hypothyroidism could affect carbohydrate metabolism and insulin secretion in adult offspring in rats. He also had some other publications on laboratory methods including the development of modified RIA and ELISA methods. He used the assays mentioned to study the plasma pancreatic and gut glucagon, GIP, insulin and gastrin responses to feeding in controls and patients with duodenal ulcer (DU) disease, who were significantly hypoglycaemic before and after breakfast compared with controls, and had significantly higher insulin, GIP and gut glucagon responses after feeding, demonstrating that the relative hypoglycaemia was enough to activate the vagus nerves and to cause peptic ulcer; he further showed that secretin provocation tests, usually used to diagnose pancreatic gastrinoma, produce marked insulin release in patients with primary hyperparathyroidism but not in controls or in patients with gastrinoma or those with DU disease.In another study, he reported that the gut hormone cholecystokinin (CCK) has a similar distribution in human brain compared with results documented for guinea pig and porcine brains. Using the gastrin assay and column chromatography to identity CCK molecules of different sizes, he also showed that in the cerebral cortex most of the CCK is in the grey matter and that in patients with Alzheimer disease there is significantly more glucagon in grey matter than in controls. He was very involved in teaching different aspects of human physiology, especially the GI tract, the endocrine system and circulation to both medical and post-graduate students, and had conducted well over fifty theses. To date he had conducted over 100 research projects, established a research laboratory and was currently working mainly on the behavior of isolated islets and isolated aorta from diabetic animals. He received many awards, including one for outstanding professor in 2011.A workaholic, Dr. Zahedi was a member of the Iranian Academy of Medical Sciences. Apart from this he held almost thirty-five other important key positions; he was a member and chairman of the physiology and pharmacology boards of the country, and was a founding member of continuing education in Iran.So many distinctive compliments come to mind when trying to eulogize Dr. Zahedi for what he was renowned and respected for, but space and words to describe him are lacking. Devout, self- effacing, affable, always the gracious gentleman, Dr. Zahedi is and will remain with us in our thoughts and prayers for a long long time.