A survey of the interactions between phosphorus (P) species and the components of calcareous soils shows that both surface reactions and precipitation take place, especially in the presence of calcite and limestone. The principal products of these reactions are dicalcium phosphate and octacalcium phosphate, which may interconvert after formation. The role of calcium carbonate in P retention by calcareous soils is, however, significant only at relatively high P concentrations – non-carbonate clays play a more important part at lower concentrations. In the presence of iron oxide particles, occlusion of P frequently occurs in these bodies, especially with forms of the element that are pedogenic in origin. Progressive mineralization and immobilization, often biological in nature, are generally observed when P is added as a fertilizer.Manure serves both as a source of subsurface P and an effective mobilizing agent. Blockage of P sorption sites by organic acids, as well as complexation of exchangeable Al and Fe in the soil, are potential causes of this mobilization. Swine and chicken manure are especially rich P sources, largely due the practice of adding the element to the feed of nonruminants. Humic materials, both native and added, appear to increase recovery of Olsen P. In the presence of metal cations, strong complexes between inorganic P and humates are formed. The influence of humic soil amendments on P mobility warrants further investigation. The mobility of phosphorus (P) in the shallow subsurface is a matter of critical importance and considerable complexity. Its importance stems from the fact that P, an essential nutrient for all plant and animal life, is often in short supply. Agricultural fertilizers and other soil amendments, such as mineral P fertilizers and animal manure, provide P that is readily available to plants. The short-term availability of P to crops is strongly influenced by biochemical processes that affect organic matter, while its long-term status is generally determined by geochemical transformations.etc. [i) and organic P (Po) species interact extensively with soil components and are subject to various chemical transformations that affect the retention of the element.The nature of P species in the shallow subsurface varies widely with location, soil type, and management system. In describing P movement in soils, workers often use operational categorizations such as "dissolved reactive P", "particulate unreactive P", etc. ,2 The abetc. . Both ini and Po in the soil [Depletion and oversupply are the two main challenges in subsurface P management. Depletion is especially serious when low input agriculture is practiced, involving land clearing and continuous cultivation that reduce both Pthe soil . Oversupthe soil ,6. Surplet al. [The monitoring and management of environmental P is predicated on accurate determinations of the element in subsurface matrices. A thorough discussion of P analysis is beyond the scope of this survey, but an excellent compilation of analytical techniques has been published under the auspices of the USDA-CREES . Sample et al. . Brieflyet al. [i (absorption at 880 nm) [(i) The Murphy-Riley (MR) colorimetric method for inorganic P analysis  , which  880 nm) .(ii) Inductively coupled plasma (ICP) spectroscopy, with either optical emission (178.290 nm)  or mass et al. in 1995 [(iii) Potentiometry with the phosphate-sensitive cobalt electrode, which was introduced by Xia  in 1995 ,15, and  in 1995 -18.31P NMR spectroscopy [et al. [i and Po. Cade-Menun et al. note that the quantitative use of 31P NMR spectra of soil and litter extracts in solution requires careful sample treatment, control of parameters, and knowledge of the species present in solution. They have published a comprehensive summary of recommendations regarding the choice of extractant, measurement of relaxation times, determination of Fe and Mn content, use of appropriate delay times, and sample temperature [Chemical identification or organic P in environmental samples is generally carried out by troscopy ,20. Spec [et al.  have pubperature .Analyses of P retention and mobilization in natural calcareous environments have shown that both adsorption and precipitation take place, although it is not always easy to distinguish between the two mechanisms. Measurement of P by any of the techniques mentioned above is usually preceded by single or sequential extractions, which often involve the solvent systems summarized in Table et al. on the interactions of P with calcite surfaces [4). Alternatively, octacalcium phosphate 2(OH)2), may be formed [10(PO4)6(OH)2), has also been found to occur [Early work by Cole surfaces  distingue formed , and hyde formed . This ise formed . A cyclito occur . The gento occur , is thatca. 2-h induction period, and then followed by precipitation [Early studies with limestone particles  suggest pitation . P-sorptet al. [3 [2O3 on calcite increases the P sorption capacity 9-fold [It is clear that surface adsorption and precipitation are major mechanisms of P retention in calcareous systems, depressing its availability after fertilizer application. Diverse results have been obtained regarding the relative roles of carbonates and oxide clays in P retention in calcareous soils. Afif et al.  found tht al. [3 ,37. The t al. [3 -40 suggey 9-fold . As the et al. [In comparing the relative importance of surface reactions and precipitation in P retention, Tunesi et al.  concludeet al. , while oet al. , are somA third retention mechanism for P, especially iron rich soils, involves occlusion in Fe oxide particles. From data obtained from single and sequential extractions with ascorbate, citrate-ascorbate, bicarbonate, dithionite, and oxalate, Torrent and coworkers ,45 conclFrom a practical standpoint it is interesting to consider how P interactions in calcareous soils compare to those in limed acid soils. In cases where substantial amounts of metal phosphates have accumulated in soils of both types – due to pedogenesis and/or fertilizer application in excess of plant uptake – calcareous soils are found to contain less surface P than limed acid soils. Ca phosphates predominate in the former, and Al and Fe phosphates in the latter. Overall, P availability to plants is greater in limed acid soils .-1, Pi is sorbed strongly, while at higher concentrations sorption energy is much lower [In unamended soils, especially those not having received manure, P leaching is generally a relatively minor problem compared to erosive losses of the element. There is, however, strong evidence that the extent of subsurface P loss is closely related to the degree of phosphorus saturation (DPS) of the soil. In terms of Olsen P, it has been suggested that below 60 mg P kgch lower , which wch lower ,50. Whench lower ,52. ThisBoth added manure or litter and native organic matter  have significant effects on subsurface P retention. Manure not only affects sorption and precipitation of P, but often contains significant amounts of the element, which is thereby – deliberately or incidentally – added to the land. Humic materials, the breakdown products of the total biota in the environment, generally are not a major source of P, but they do have a mobilizing effect on it in the subsurface. The use of extrinsic humates, especially leonardite humic acid, for soil improvement has experienced an upswing in recent years .2PO4) fertilizer appear to contribute to different P pools [2 extractable P and Mehlich-3 P, while manure  has a greater effect on modified Morgan P, as well as other types of P.The application of manure is widely practiced to increase the productivity of soils that contain inadequate levels of organic carbon. The effects of manure on P availability in various soils has been widely studied, and the general conclusion has been that it is a source of P; interacts with soil components in a manner that increases P recovery by crops; and enhances the effectiveness of inorganic P fertilizer. P added from manure and other sources, however, tends to become less available to plants with the passing of time . As ment P pools . The lat-1) annual manure applications are considered to pose a risk of ground water contamination, as the total P concentration increases with soil depth. The ability of acid soils to retain added P after long-term manuring, is generally low. It has been reported that manure applications have a greater effect on the retention of Pi than the retention of Po [Alkaline soils subject to long-term manure amendments have been shown to accumulate substantial quantities of P, with 50–66% in plant available forms [on of Po .The affinity constants and sorption capacities of soils for P are reduced by organic amendments, especially manure. This can be due to competition for P fixation sites by organic acids, and/or the complexing of exchangeable Al and Fe by components of manure -60. The vide infra) on the one hand, and root exudates on the other hand. It is well established that cover crops such as white lupin  form cluster roots in response to P deficiency, and that these root systems are efficient producers of succinate, citrate, and malate [Parallels may be drawn between the P mobilizing effects of manure and humic materials  P. The bulk of this (75–85%) is Plubility . Short tse notes , howeveret al. [o by phosphatase enzymes, especially phosphomonoesterase, can proceed over periods extending to 20 weeks in soils amended with chicken manure. The orthophosphate content of the soils increases during mineralization, while Po decreases. The sorption of Po  is positively correlated with both organic matter and Fe and Al content of the soil [The chemical identification of P species in manure is of considerable practical importance, since the exact nature of P is a major determinant in the subsurface retention of the element after manure application. Work by Crouse et al.  has showthe soil . Especia2O and NaHCO3, while much of the soil P requires NaOH and HCl. This is related to the fact that soils contain ca. 15 times as much Al, and 10 times as much Fe as manure, while manure tends to have higher Ca and Mg contents. Rapid evaluation of plant-available P clearly is a desirable feature of subsurface analysis, and He and coworkers have introduced a shortcut in the assessment of contributions from manure amendments. They suggest that a single P extraction from dairy manure with a 100 mM acetate buffer at pH 5.0 equals the combined H2O, NaHCO3, and NaOH extractions [The physico-chemical characteristics of manure differ from those of soil, and the use of sequential extractions in manure analysis needs to be carefully evaluated.  A major ractions .o species in manure, such as phospholipids and simple phosphate monoesters, can, despite their relatively low abundance, become a major P component in runoff [3 for readily soluble P, followed by 0.5 M NaOH/50 mM EDTA for more recalcitrant P. Recoveries were superior to those obtained with Hedley and NaHCO3/HCl procedures.Turner and Leytem caution that the presence of organic P in the HCl extract of the Hedley fractionation  procedurn runoff . Turner n runoff , involvii in the H2O extractable fraction of dairy manure is correlated with total P , while g manure . In cont2(SO4)3·14H2O or KAl(SO4)2·12H2O). Of these, alum has proven to be most effective, with the added benefit that it also prevents the loss of ammonia [Chicken manure and swine slurry are apt to provide readily mobile (water soluble) P to soil, which can lead to runoff and eutrophication problems. For this reason, some effort has been expended at reducing the mobility of P in these types of manure and litter. Chemical additives that have been used for this purpose ,84, incl ammonia ,86 and w ammonia ,88.et al. [31P NMR to elucidate the nature of the immobilized P species. This proved to be a difficult task, involving many unresolvable Pi and Po species. It was noted that no crystalline aluminum phosphate species were present.Al-associated P accounts for some 40% of total P in alum amended materials, with about 20% of this being drawn from Ca-phosphate phases. This decreases by about half when alum is added to poultry litter. Hunger et al.  have usei.e. the release of P under the influence of dissolved humic materials, Delgado et al. [Humic and fulvic acids comprise a wide variety of organic materials that are present in all agricultural soils. Their effects on plant growth and nutrition are well documented, ,91 and to et al.  have pubi and humic materials is predicated on the presence of metal ions that act as cationic "anchors", allowing anionic humates and phosphates to associate [A recent investigation indicates that strong interactions between Pssociate . StabiliMuch has been learned about P mobility in calcareous media over the last five decades, but some gaps in understanding remain. Many of these occur at the molecular level of P interaction with subsurface species, including the detailed mechanism of P desorption under the influence of organic species. The role of humic materials in P mobilization is another area of research that has been given relatively little attention and is a potentially fruitful area of study. The use of humates as soil amendments presents an especially interesting case. The practice is gaining popularity – as borne out by the existence of more than 70 purveyors of these "nonconventional soil additives" in the U.S. alone  – but no


Hotez et al. argue that achieving success in the global fight against HIV/AIDS, tuberculosis, and malaria may well require a concurrent attack on the neglected tropical diseases. The age of hypocrisy has been succeeded by that of indifference, which is worse, for indifference corrupts and appeases: it kills the spirit before it kills the body. It has been stated before, it bears repeating: the opposite of love is not hate, but indifference. —Elie WieselTIME Global Health Summit, branded by Bono as the “Woodstock of Global Health” .The last five years have witnessed increased efforts by G8 nations and United Nations agencies to improve the health of the world's 3 billion people living on less than US$2 a day. Most of this attention has focused on efforts to intensify resources for fighting the three most devastating diseases: HIV/AIDS, tuberculosis, and malaria. Together, these “big three” account for a staggering 5.6 million deaths and the loss of 166 million disability-adjusted life years (DALYs) annually , and schistosomiasis . CysticeThe burden of disease resulting from the neglected tropical diseases is huge. As shown inAscaris lumbricoides,Trichuris trichiura, and the hookworms,Necator americanus andAncylostoma duodenale) and the two schistosomes (Schistosoma haematobium andSchistosoma mansoni) are highly endemic to sub-Saharan Africa, where they adversely affect childhood growth and physical fitness ),The President's Emergency Plan for AIDS Relief (http://www.avert.org/pepfar.htm),The Bill and Melinda Gates Foundation (http://www.gatesfoundation.org), supporting the development of new technologies for disease control.


Cells react to changing intra- and extracellular signals by dynamically modulating complex biochemical networks. Cellular responses to extracellular signals lead to changes in gene and protein expression. Since the majority of genes encode proteins, we investigated possible correlations between protein parameters and gene expression patterns to identify proteome-wide characteristics indicative of trends common to expressed proteins.Drosophila melanogaster life span, and Saccharomyces cerevisiae cell cycle.Numerous bioinformatics methods were used to filter and merge information regarding gene and protein annotations. A new statistical time point-oriented analysis was developed for the study of dynamic correlations in large time series data. The method was applied to investigate microarray datasets for different cell types, organisms and processes, including human B and T cell stimulation, We show that the properties of proteins synthesized correlate dynamically with the gene expression profile, indicating that not only is the actual identity and function of expressed proteins important for cellular responses but that several physicochemical and other protein properties correlate with gene expression as well. Gene expression correlates strongly with amino acid composition, composition- and sequence-derived variables, functional, structural, localization and gene ontology parameters. Thus, our results suggest that a dynamic relationship exists between proteome properties and gene expression in many biological systems, and therefore this relationship is fundamental to understanding cellular mechanisms in health and disease. Cells react to changing intra- and extracellular signals by dynamically modulating complex biochemical networks, and cellular responses to extracellular signals lead to changes in gene and protein expression. These processes can be monitored using genomics and proteomics methods. A number of supervised and unsupervised clustering techniques are routinely applied to classify and group genes based on their expression profiles . While tSince the majority of genes encode proteins, we investigated possible correlations between protein-related properties and gene expression patterns to identify proteome-wide features indicative of trends common to expressed proteins. For example, because the cytoplasm, nucleus and extracellular space have different physicochemical properties, such as pH, ionic composition, and protein concentration, the properties of the proteins that are targeted to different cellular compartments are also different. Because there is variation in the specific proteins that comprise the various proteomes, it is intriguing to hypothesize that cellular signaling leads to significant changes in the protein properties of cells. This idea is supported by studies of the relationship between the overall properties of proteins and their amino acid composition, which has been correlated with protein surface properties , subcellDrosophila melanogaster life cycle data [A number of microarray datasets for several different cell types and organisms were analyzed to study possible transcriptome-proteome correlations. Expression studies have revealed certain correlations between genome-related features and coexpressed genes, including co-localization -4 and thcle data .T cell receptor (TCR) activation on the surface of T cells is essential for mounting an adaptive immune response against viruses and microbes. The TCR is a multiprotein complex that activates a large number of signaling pathways . Both thTo study transcriptome-proteome correlations, we introduced time point-oriented analysis. Protein sequence-derived parameters were correlated to gene expression separately at each time point. Time point-oriented clustering (TOC), in which the expression of genes at each time point is clustered separately, was used for other features. Fig. The 114 variables at six time points in two data sets (CD3 and CD3/CD28 costimulation) yielded 1,368 correlation tests. The huge amount of information was organized according to time points and variables, clustered using the Self Organizing Map (SOM) method and visualized as in Figs. Of particular interest are the characteristics that indicated trends in more than one data set or at more than one time point. Of the 75 and 102 significant results in the CD3 stimulation and CD3/CD28 costimulation experiments, respectively, 44 (25%) were common to both /total parameter, correlating positively in both the CD3 and CD3/CD28 costimulation datasets. The aliphatic index, which measures the proportion of aliphatic residues , correlated negatively in the CD3 dataset. The individual aliphatic residues, however, generally gave positive correlations, with the exception of L and V. The frequency of S and T correlated negatively with gene expression in both datasets. The data also suggest that aromatic residues do not generally covary with gene expression. The ratio of polar and large, negative, positive, as well as nonpolar and small residues correlated positively in both datasets. The effect was stronger in the costimulation dataset. Nonpolar residues correlated negatively. Overall hydropathy parameters correlated negatively with expression levels, although this type of correlation varied across time points, due most likely to differences in the parameters contributing to the hydropathy scales. The metabolic cost of amino acid production has been previously evaluated; e.g., for Of particular interest are the characteristics that yielded negative correlations at early time points and positive correlations at late time points, and vice versa. The trend is from a negative towards a positive correlation for the ratio of positive amino acids and (R+K)/total parameters in the CD3 dataset, and D in the costimulation dataset. Serine showed the opposite trend in the CD3 dataset. The proportion of neutral residues and oxidoreductase activity correlated negatively at early time points, but only in the CD3 dataset. The ratio of nonpolar and small amino acids correlated negatively at early time points in the CD3 dataset and positively only at late time points in the costimulation dataset. These results clearly indicate a strong correlation between gene expression and the amino acid composition of protein products. In particular, polar and charged residues correlate strongly, and there is a biased occurrence of hydrophobic and aliphatic residues. Interestingly, leucine and isoleucine, which are hydrophobic structural isomers, had opposite correlations.Protein molecular size and weight are weak indicators, because only MWRES (molecular weight per residue) correlated significantly in the costimulation dataset. Of the volume-related parameters, nonpolar and polar volume correlated negatively, and Polfrac max correlated positively in costimulated cells. Protein flexibility correlated positively in the CD3 dataset at one time point. The most flexible residues, G and A, did not correlate with the transcriptome (except for A at two time points).SCOP  containsFifty ontology classes had a large number of entries, 26 of which correlated significantly with expression Fig. . Many ofIntracellular proteins correlated positively towards the end of the time series in both datasets. Membrane proteins in general and those integral to the plasma membrane were significantly underrepresented at several time points. Based on the prediction data, extracellular proteins correlated negatively with gene expression in the CD3/CD28 dataset.Our results are somewhat related to previous analyses of the yeast proteomes ,29, dataThe previous yeast study  was aime2+-related signaling, and NF-κB pathways. Subsequently, transcription factors became activated, and a significant positive correlation was observed at time point 3 in CD3-stimulated cells. The enriched transcription factors included early growth response factors 1, 2 and 4, NFAT, NF-κB, Jun, Fos, B cell translocation gene, and interferon regulatory factor. Thus, all the major transcription factor components active in TCR signaling were present . In agreement with these results, transcription factors expressed in lymphocytes have highly regulated expression patterns [TCR stimulation activates several signaling pathways ,32,33, apatterns .We analyzed a number of additional datasets to address whether the associations seen in the T cell data analysis represent sporadic observations that are cell type-specific. Dynamic covariation occurred between gene expression profiles and a number of proteome characteristics describing amino acid proportions and physicochemical properties in all the investigated cell types and organisms, suggesting that our T cell data is representative of cells in general.The B cell dataset ,21 repreProline and R/(R+K) were significantly enriched during several consecutive late time points, and neutral and small residues were enriched in the last time point Fig. , whereasS. cerevisiae cultures synchronized by three independent methods: α factor arrest, elutriation, or arrest of a CDC15 temperature-sensitive mutant [Microarrays have been used to study gene expression in e mutant . Expresse mutant .Several significant correlations are apparent in Fig. At the early time points, the correlations are positive  for F, G, I, L, V, W, Y, hydropathy values, as well as for ratios of nonpolar and large, nonpolar, and neutral amino acids. Correlations are mainly negative for E, K, (R+K)/total, rho, and ratios of positive, and polar and small residues. The correlation of V to gene expression is consistent with a previous yeast study ; howeverLarge numbers of correlations occur in the CDC15 and CDC28 experiments, in which the largest fraction of significant MIPS functional classifications was observed in a previous study . This imD. melanogaster study was performed to follow development and gene expression in a multicellular model organism. In this dataset [The  dataset , the traD. melanogaster data, in that once a significant correlation occurs it remains for a prolonged period .There are dynamic correlation patterns between gene expression and proteome properties during the embryonic period and especially the pupae period Fig. . CorrelaThe data contain two major transitions of correlations that do not coincide with developmental stages Fig. . The firThe larval and adulthood data show stable and constant patterns, in which correlations occur between gene expression and a large number of parameters, including a negative correlation for charged and polar residues, their ratios, Polfrac max, rho and flexibility. Positive correlations are seen for aliphatic, aromatic and hydrophobic amino acids, aliphatic index, and hydropathy parameters. Contrary to the observations for T cells, I and L show similar trends. As noted above, the patterns of positive and negative correlations are almost mirror images when comparing the first half of the embryo stage with the two constant regions . Almost all protein properties have statistically significant correlations; the exceptions are N, T and the ratio of neutral and small residues, for which significant correlations occur sporadically.Several gene ontologies (17 classes) have significant correlations. Six ontologies are related to gene expression/DNA binding. They all have positive correlations during the first half of the embryonic stage and negative correlations thereafter. Nuclear localization follows the same pattern. The correlation motifs for the other ontologies are less clear. Oxidoreductase activity correlates negatively at several time points during the embryonic stage and correlates positively at other time points throughout the experiment. The signal transduction ontology has a similar overall pattern, but only in the pupa and adult females.D. melanogaster life cycle datasets have obvious recurring cycles of enrichment and underrepresentation of different properties. The magnitudes of underrepresentation and enrichment are proportional to gene expression in all experiments. These observations along with our findings related to the functional and structural properties of proteins as well as MIPS classification in earlier studies [D. melanogaster dataset.Strong correlations between transcriptome and proteome characteristics appear in all the datasets we studied. One could not expect to obtain similar results from the different studies, which differed with respect to both the processes  and proteins produced. The strength of the correlations is apparently associated with the intensity of gene expression. Thus, proteins having significantly altered expression do not have a random distribution of physicochemical properties. In particular, the yeast and  studies ,21 suggeD. melanogaster, indicating that the dynamic correlations between proteome-related parameters and gene expression likely represent a general paradigm. Since the association is strong and is observed at many levels, cells and organisms, and appears to be a widespread phenomenon, it likely reflects fundamentally important biological processes.Similar, although not identical correlations were obtained when analyzing time series datasets for human T and B cells, yeast, and Only a few studies of system-wide transcriptome-proteome correlations have been published, including human heart  and platProteome-wide isoelectric points and molecular masses, the two properties used for separation in 2D gels, have been analyzed for 103 organisms, including bacteria, archae and eukaryotes . The comThere are presumably several reasons for the observed enrichment/depletion of protein properties, which might be related, for example, to cellular processes involved in changes in metabolism and signaling, localization of proteins within cells and compartments, and complexes and interactions formed between proteins. There are indications for the organellar enrichment of proteins having certain types of properties; for example, the need for positive charge in DNA-binding proteins (including histones). In certain disease conditions, the overall amino acid composition of proteins changes in cells, organs, or body fluids ,42. Thus.Gene expression data were derived from peripheral T cells subjected to seven treatments: mock (untreated) cells; CD3- or CD28-stimulated cells; CD3 and CD28-costimulated cells; and cells treated with ionomycin along with phorbol 12-myristate 13-acetate (PMA), phytohemagglutinin, or FK506. Data were acquitted at six time points up to 48 hr . We did D. melanogaster microarray data set, the transcriptional profiles were investigated throughout the life cycle [ from file Arbeitman.SOMtables.zip. Gene expression information was from .The B cell dataset for genes involved in maturation in anti-immunoglobulin M-stimulated Ramos cells indicated that close to 1,500 genes had significantly altered expression, at least at one time point ,21. Thesfe cycle . RNA expfe cycle . The dat and .A total of about 600 genes had altered gene expression patterns in yeast cultures synchronized by four independent methods  . We alsoD. melanogaster protein sequences, and NCBI genome sequences were used for yeast.We used numerous bioinformatics methods to filter and merge information regarding gene and protein annotations in a number of databases and further calculated and predicted a large number of characteristics for each gene/protein. A dedicated database  constructed for gene and protein information was extensively used for annotations and sequence identification. To identify the corresponding proteins, we used Locus Link ID numbers of ESTs and genes to retrieve UniProt sequences. FlyBase  was usedPhysicochemical parameters and amino acid proportions were directly calculated from protein sequences. Amino acids were further investigated by analyzing the proportions of different groups of residues, namely positive , negative , neutral , polar , and nonpolar . Combinations of characteristics were also investigated, namely neutral and small , nonpolar and large , nonpolar and small , polar and large , polar and small . In addition, the ratio of Glx  and Asx  to total amino acids, the ratio of R to R plus K, and the ratio of R and K to total amino acids were calculated.Several parameters for physicochemical features described in the literature were calculated. The aliphatic index is defined as the relative volume occupied by aliphatic side chains  and was calculated as followsAI = X(Ala) + 2.9 × X + 3.9 × (X(Ile) + X(leu)),rho is the ratio of polar to non-polar volumes. Nonpolar side chains (NPS), the frequency of nonpolar side chains, was calculated according to Waugh's definition by counting W, N, T, F, P, L, and V residues and expressing the sum as a fraction of the total number of residues [where X(Ala), X, X(Ile), and X(Leu) are mole percents for the amino acids . Residueresidues . MWRES iresidues  was calci nis number of residue i, and Hi is the hydrophobicity value of the residue. Four different hydropathy scales [where y scales -51 were POLFRAC_MAX (polar fraction) in an extended chain conformation is calculated as:NRES is the total number of residues in the protein and POLFRAC_MAX is in units of e, the electronic charge, SURF is maximal accessible surface area, and POLSURF is polar surface area [ program iep. In vivo half-life of proteins was calculated according to [where ace area . Moleculrding to .Average flexibility ,55 for e and Bnc is the flexibility parameter of the residue in position i.where D. melanogaster [For the T cell experiment , we idennogaster , 2,976 a, which yielded predictions for four categories . Transmembrane regions were further predicted using the TMHMM server [. To assign each protein to a compartment, we used the SubLoc reliability index and the length of the predicted transmembrane region. If the value of the SubLoc prediction parameter was ≥ 5, the assignment was accepted for the most accurate prediction. In cases where the parameter was between 2 and 4, the length of the transmembrane stretch(es) was taken into account. If the transmembrane region was longer than 36, the protein was predicted to be membrane associated. In cases where the SubLoc prediction value was 1 and the transmembrane region was ≥ 18 residues (the average length of a transmembrane α-helix), the protein was predicted to be membrane spanning. We made predictions in T cell data for 443 proteins, 177 of which were transmembrane, 167 nuclear, 67 extracellular, 7 mitochondrial, and 25 cytoplasmic. Predictions were made for 224 proteins in the B cell data.Subcellular localization of proteins was predicted using SubLoc software , which yM server . To assiGene ontology  informat, were used to search SCOP-derived domains [-6 as a limit. SCOP information was available for 2,022 entries. The limit of e-values was set so that false positives were unlikely. Moreover, we compared a number of findings with those acquired using the InterProScan sequence searching service [Hidden Markov Models (HMMs), downloaded from Superfamily , were us domains  against  domains , setting service , which yDrosophila dataset. To estimate the significance of the observations, a permutation test was performed. One-thousand permutations were calculated for each parameter at each time point in the B cell, T cell, and yeast datasets, and 100 permutations were calculated in the D. melanogaster dataset. To be regarded as significant, the p value had to be ≤ 0.05, both in the correlation and permutation calculations.To monitor dynamic covariations between a number of proteome parameters and gene expression levels along the time series, we applied the Spearman linear correlation test. The schema of the analysis is presented in Fig. For TOC, the expression of genes at each time point are separately clustered. For the actual clustering, we used the SOM method, which organized the clusters according to the shapes of the expression profiles of genes. During the method development, we also monitored the clustering process visually; SOMs served as ideal visualization tools. Following TOC, the Spearman correlation was used to test the linear correlations between the medians of gene expression and protein variables in the corresponding clusters.TOC was an essential step in facilitating the analysis of functional and structural attributes  because, due to the nature  and scarceness of some of these data, a clustering measure was required before conducting the correlation test. Furthermore, all correlation coefficients could be compared by unifying the sample sizes with a fixed number of clusters (20) for each of the different types and data samples.Since the functional and structural information variables were categorical , we conducted correlation tests between the medians of the clusters of expression levels and medians of the ratios (observed/expected in each cluster) of the sequences attributed to the functional or structural variables. The expected number of a given ontology, subcellular localization, or structural variable for each cluster was calculated based on the size of the cluster and the total number of occurrences of the variable.Drosophila datasets, respectively.The correlation coefficients and p values for correlation and permutation tests are in Additional files To provide an intelligible report on the very large correlation analyses, we introduced a new type of visualization. Correlation matrices were formed in which columns and rows represent time points and variables, respectively. Each cell represents a correlation coefficient  between the expression levels of the time point and the values of the variable. Red and green represent positive and negative correlations, respectively. The intensities of the colors are relative to the absolute value of the correlation coefficients. A significant red-colored proteome variable implies an increase in production  compared with the changes in expression of other proteins, which implies the enrichment of the property at that time point. Likewise, a significant green-colored variable implies a reduction in the production of proteins either associated with or containing lower proportions of the property . To determine which variables covary in a similar manner with expression levels over time, the matrix was clustered using SOMs. was used for clustering. We developed the software for all other analyses and calculations as well as for all visualizations.Script from CPAN MTAS participated in the statistical analysis, preliminary program development, and drafted the manuscript. MS carried out the data mining and statistical analysis, developed computer tools, and drafted the manuscript. TOL participated in data analysis and developed data mining, analysis and visualization tools. MV conceived of the study, participated in its design and coordination, and drafted the manuscript. All authors read and approved the final manuscript.CD3-stimulated T cellsClick here for fileCD3/CD28-costimulated T cellsClick here for fileB cellsClick here for fileSynchronized yeast cellsClick here for fileDrosophila life cycle datasetClick here for file


Previous studies have linked smaller kidney dimensions to increased blood pressure. However, patients with Balkan Endemic Nephropathy (BEN), whose kidneys shrink during the course of the disease, do not manifest increased blood pressure. The authors evaluated the relationship between kidney cortex width, kidney length, and blood pressure in the offspring of BEN patients and controls.102 offspring of BEN patients and 99 control offspring of non-BEN hospital patients in the Vratza District, Bulgaria, were enrolled in a prospective study and examined twice (2003/04 and 2004/05). Kidney dimensions were determined using ultrasound, blood pressure was measured, and medical information was collected. The parental disease of BEN was categorized into three groups: mother, father, or both parents. Repeated measurements were analyzed with mixed regression models.In all participants, a decrease in minimal kidney cortex width of 1 mm was related to an increase in systolic blood pressure of 1.4 mm Hg (p = 0.005). There was no association between kidney length and blood pressure. A maternal history of BEN was associated with an increase in systolic blood pressure of 6.7 mm Hg (p = 0.03); paternal BEN, +3.2 mm Hg (p = 0.35); or both parents affected, +9.9 mm Hg (p = 0.002). There was a similar relation of kidney cortex width and parental history of BEN with pulse pressure; however, no association with diastolic blood pressure was found.In BEN and control offspring, a smaller kidney cortex width predisposed to higher blood pressure. Unexpectedly, a maternal history of BEN was associated with average increased systolic blood pressure in offspring. There is a large body of evidence that patients with Balkan Endemic Nephropathy (BEN), in contrast to other chronic kidney diseases ,2, do noWe conducted a prospective study in the Vratza District of Bulgaria, a rural area, and measured the kidney cortex width, kidney length, and blood pressure of the offspring of BEN patients and control offspring at two time points, one year apart. We hypothesize the following:- A smaller kidney cortex width is related to a higher average systolic blood pressure and a higher pulse pressure.- An offspring with parental history of BEN has a higher average systolic blood pressure and a higher pulse pressure.In the period of October, 2003, to April, 2004, we recruited adult offspring (102 study subjects) whose father and/or mother were included in the Vratza Hospital registry of BEN patients in 2001 and who resided in one of three communities . The diagnosis of BEN in the parent generation is based on three groups of criteria: epidemiological, clinical/laboratory, and pathological/anatomical /3600)To test the hypothesis that offspring of BEN parents have increased blood pressure, we grouped parental disease status into three groups: mother, father, or both parents affected. None of the comparison offspring fell into any of these groups (reference). The effect of cortex width and parental BEN status was tested in the same model.In order to estimate the association of minimal kidney width and parental history of BEN with blood pressure based on repeated measurements, we applied linear mixed models. Measurements of the two investigations are not independent and mixed models allow adjusting for within-participant effects . The mixTo compare the agreement between measurements of continuous variables in 2003/04 and 2004/05 we used the intra-class correlation coefficient (ICC). This is the between-subject minus the within-subject variance divided by the sum of the two variances. The ICC quantifies the proportion of total outcome variance that is due to inter-individual variation.Of the 201 participants in the first investigations (2003/04), 189 participated in the follow-up . Two were deceased, one moved out of the area, two could not be contacted, and 7 decided not to participate further. Of the 12 lost, 9 were offspring of BEN patients. Potential risk factors show similar distributions in each of the two investigations . When analyzing blood pressure differences while controlling for confounders, cortex width and parental BEN status had significant effects on systolic and pulse pressure, but not on diastolic blood pressure. A decrease in the minimal kidney cortex of 1 mm was linearly related to an increase in systolic blood pressure of 1.4 mm Hg (p = 0.005) and an increase of pulse pressure of 1.1 mm Hg (p = 0.002).Figure A maternal history of BEN was associated with an increased systolic blood pressure in offspring of 6.7 mm Hg (p = 0.03), and when both parents had BEN, an increased systolic blood pressure of 9.9 mm Hg (p = 0.002). There was no statistically significant effect of a paternal history of BEN.To investigate whether the use of antihypertensive medication had distorted our findings, we repeated the model excluding those observations with antihypertensive drug use. This reduced the number of repeated observation from 388 to 322 (176 subjects), but did not change our findings.Our findings show that a decrease of the minimal kidney cortex width of 1 mm was related to an increase in systolic blood pressure of 1.4 mm Hg. There was no association between kidney length and blood pressure. A maternal history of BEN was associated with a significant increase in systolic blood pressure of 6.7 mm Hg, paternal BEN with a non-significant increase of 3.2 mm Hg, and both parents affected, a significant increase of 9.9 mm Hg. The lack of association of either kidney cortex width or a parental history of BEN with diastolic blood pressure may either stem from the larger individual variance of this parameter or may indicate that there is no such link.In a study with healthy volunteers in England, Raman et al. did not identify a clear pattern between measurements of the right and left kidneys and blood pressure . In anotFor adult Aboriginals in Australia, Singh et al. found that a 10 mL decrease in kidney volume was associated with a 0.5 mm Hg increase in systolic blood pressure . The stuMackenzie et al. posited the hypothesis that the total nephron supply at birth is a factor in determining a susceptibility to increased blood pressure . Given tHence, a maternal history of BEN may have both a direct and an indirect effect on blood pressure in offspring. Indirectly, maternal BEN seems to decrease kidney cortex width, which in turn increases blood pressure in adult offspring. Directly, maternal BEN seems to initiate regulations likely to raise the average blood pressure in affected offspring. However, hypertension is not a predominant feature in BEN offspring. Thus, it is unlikely that the reduction in kidney cortex width in BEN offspring results from hypertension.Renal mass is considered to contribute directly to the development and maintenance of hypertension ,19,28. AHowever, in BEN patients, whose kidneys shrink, the lack of increased blood pressure has been considered to be a typical characteristic before the disease reaches its final stage -9. The rSince our data is based on adult offspring, we are uncertain about the future development of blood pressure in this group. There are three possibilities: (I) Offspring of BEN patients will not develop BEN but maternal BEN may lead to subclinical changes such as an increase in average blood pressure. An argument against this option is that we have found reduced kidney sizes and increased protein excretion in the BEN offspring, which may be considered as early signs of BEN . (II) BEWe found that adult offspring of BEN patients have an average increased blood systolic pressure and pulse pressure, determined in two repeated measurement one year apart. This is contrary to reports in BEN patients, who do not have this characteristic. Surprisingly, the increase was related to a maternal history of BEN, or a history of BEN in both parents, but not to a paternal history alone. Increased blood pressure was also associated with a reduction of the minimal width of the kidney cortex, but not with a reduction in kidney length. In our understanding, these findings emphasize the role of maternal priming in determining susceptibility for increased blood pressure. Balkan endemic nephropathy may serve as a model to better understand the development of both kidney disorders and their related blood pressures.The author(s) declare that they have no competing interests.PSD developed and designed the study and helped write the report. SDT assisted in analyzing the data, literature review, and writing the discussion. RBG conducted the analyses of lead in blood, and assisted in writing the discussion. VAS conducted the clinical examinations of the study subjects and contributed to the interpretation. AGB conducted the ultrasound examinations of the study subjects and contributed to the interpretation. WJK helped design the study and conducted the statistical analysis and worked on the manuscript. All authors read and approved the final manuscript.The pre-publication history for this paper can be accessed here:


Little information is available on leptin concentrations in individuals with IGT. This study aims to determine and correlate leptin levels to anthropometric measures of obesity in pre-diabetic, (IFG and IGT), type 2 diabetic and normoglycaemic Saudis.308 adult Saudis  participated. Anthropometric parameters were measured and fasting blood samples taken. Serum insulin was analysed, using a solid phase enzyme amplified sensitivity immunoassay and also leptin concentrations, using radio-immunoassay. The remaining blood parameters were determined using standard laboratory procedures.Leptin levels of diabetic and pre-diabetic men were higher than in normoglycaemic men . In females, leptin levels were significantly higher in pre-diabetic subjects (14.09 [2.8–44.4] ng/mL) than in normoglycaemic subjects (10.2 [0.25–34.8] ng/mL) (p = 0.046). After adjustment for BMI and gender, hip circumference was associated with log leptin (p = 0.006 with R2 = 0.086) among all subjects.Leptin is associated with measures of adiposity, hip circumference in particular, in the non-diabetic state among Saudi subjects. The higher leptin level among diabetics and pre-diabetics is not related to differences in anthropometric measures of obesity. Saudi Arabia, a Middle Eastern country with a population of 22 million, has undergone significant economic and cultural changes over the past thirty years. Approximately 60% of the population are urbanised and have adopted a 'Westernised' lifestyle in terms of diet and physical activity . ObesityIncreased risk of CHD in subjects with IGT is supported by data from the Honolulu , ChicagoDiabetes increases the risk of developing cardiovascular disease by 2–3 times and increases by as much as 50% the risks of non-cardiovascular mortality associated with this condition -15. Thisper se which is regarded as a pathogenic mechanism in human obesity. Leptin concentrations vary widely among individuals with similar fat mass, indicating other possible factors for its determination ) than in the controls.2 = 0.086 and p value of 0.006 . Even after adjustment for BMI, there was a positive association between log leptin and hip circumference with an RInsulin resistance is the common factor in a range of risk factors for atherosclerosis, specifically hypertension, dyslipidemia and abnormal glucose metabolism. As has been shown in other populations -29, the Data were presented according to gender, since it is already an established fact that leptin levels are significantly higher in women than in men. There are several possible explanations for the difference. One is that females have more adipose tissue than males, but a growing literature indicates that estrogen, especially at higher levels, will stimulate the production of leptin, whereas androgens will suppress the levels of leptin . In thisSerum leptin concentration may contribute to the risk of CVD by altering lipid metabolism and contributing to hypertension via the activation of the sympathetic nervous system and increasing renal sodium re-absorption ,43. HoweA family history of diabetes was found to have a negative association with leptin in pre-diabetic patients; this confirms a previous study which reported that healthy lean non-diabetic Asian Indians are more insulin resistant than other ethnic groups, despite similarities in their living environment . Waist cThe significant correlation of leptin to selected anthropometric measurements of obesity is confirmed in non-diabetic Saudi Subjects. In those with diabetes, this relationship is lost, reflecting the effect of other factors, including hyperinsulinaemia and the activation of sub-clinical inflammation.The author(s) declare that they have no competing interests.NA for the concept and design as well as statistical analysis; OA for the drafting and revising of the manuscript; KA and AJ for the acquisition and interpretation of data; MM and MK for the screening and collection of data; KS for the concept and final revision of the manuscript. All the authors have read and approved the final version.


The effects of TZDs on cancer risk in humans needs to be resolved as these drugs are prescribed for long periods of time in patients with diabetes.Peroxisome proliferator-activated receptors (PPARs) have emerged as important drug targets for diabetes. Drugs that activate PPARγ, such as the thiazolidinediones (TZDs), are widely used for treatment of Type 2 diabetes mellitus. PPARγ signaling could also play an anti-neoplastic role in several A total of 1003 subjects in community practice settings were interviewed at home at the time of enrolment into the Vermont Diabetes Information System, a clinical decision support program. Patients self-reported their personal and clinical characteristics, including any history of malignancy. Laboratory data were obtained directly from the clinical laboratory and current medications were obtained by direct observation of medication containers. We performed a cross-sectional analysis of the interviewed subjects to assess a possible association between cancer diagnosis and the use of TZDs.P = 0.04). This association was particularly strong among patients using rosiglitazone , and among women .In a multivariate logistic regression model, a diagnosis of cancer was significantly associated with TZD use, even after correcting for potential confounders including other oral anti-diabetic agents (sulfonylureas and biguanides), age, glycosylated hemoglobin A1C, body mass index, cigarette smoking, high comorbidity, and number of prescription medications (odds ratio = 1.59, These data suggest an association between TZD use and cancer in patients with diabetes. Further studies are required to determine if this association is causal. Factors affecting cancer incidence in the diabetic population are diverse and complex. Diabetes is a risk factor for colon, pancreas, breast, endometrium and liver cancer in women. In men, although diabetes it is a risk factor for bladder cancer ,2, it isThe increasing prevalence of diabetes has intensified the search for new therapies. Orally administered anti-diabetic agents  can be used alone or in combination with other oral agents or insulin . Over thin vivo [The TZDs are attractive options in the therapy of diabetes because they decrease insulin resistance -10. In ain vivo , lung cain vivo ,16, thyrin vivo , hepatocin vivo  and othein vivo . Seargenin vivo  demonstrin vivo . As suchin vitro and in vivo models that do not take into full account the metabolic characteristics of patients with diabetes. Recently, a population-based report showed that TZDs were associated with reduced risk of lung cancer in diabetic patients [in vitro studies [Evidence of both pro-carcinogenic and anti-tumorigenic effects of PPARγ agonists have been mainly studied patients  in agree studies ,16. HereThis study is part of a larger project, the Vermont Diabetes Information System (VDIS), a study of 8855 adults with diabetes in primary care practices . The subSubjects completed a questionnaire at home and then were visited by a trained research assistant who reviewed the questionnaire responses, assisted the subject with any missing or unacceptable responses, reviewed the subject's medications, and measured their height and weight using a portable stadiometer and scale. Race, education, income, marital status, functional status, smoking, alcohol consumption and comorbid conditions were obtained by questionnaire. Prior to the interview, patients were instructed to gather all current medications, including over the counter preparations, for review by the research assistant. The medication list was ascertained by direct observation of the medication container with recording of the drug name, dose, frequency and route of administration. Duration of therapy was not recorded.We used the SF-12 health survey to determine physical and mental functional status. Results were scored and normalized using designed algorithms and expressed in terms of the Physical Component Summary (PCS) and the Mental Component Summary (MCS) . To deteThe interviews occurred between July 2003 and March 2005. Most laboratory data were obtained from the patients' local clinical laboratories, which all use the same Diabetes Control and Complications Trial/Epidemiology of Diabetes Interventions and Complications high performance, liquid chromatography (HPLC) method for the determination of glycosylated hemoglobin (A1C). Less than 1% of A1C tests were performed using the Bayer DCA 2000 immunoassay point of care instrument, which has been shown to compare favorably with the HPLC method .The research protocol was carried out in compliance with the Helsinki Declaration and was approved by the Committee on Human Research of the University of Vermont. The interviewed subjects provided written informed consent. The full study protocol and variables and the medication profiles of the subjects have been previously reported ,26.2), alcohol drinking (yes/no), cigarette smoking (yes/no), income (in eight ordered categories), duration of diabetes in years, high comorbidity (two or more comorbid conditions), number of prescription medications, number of anti-diabetic drugs, physical function status and mental function status. To reduce the number of variables in the final model, we excluded potential confounders that were associated with the outcome in univariate analyses with P > 0.2. Such weak association argues that the variable is unlikely to be a confounder. However, variables representing use of TZDs, sulfonylureas and biguanides were included regardless of their P-values. Because diabetes is associated with an increased incidence of breast and endometrial cancers and a reduced incidence of prostate cancers [We performed a cross-sectional analysis of the interviewed subjects at the time of their enrollment in the VDIS trial. We explored the association between cancer and the use of oral agents, using logistic regression with history of cancer as the outcome variable and the use of TZDs as the primary predictor variable. We then adjusted for possible confounding by social and clinical factors. Potential confounders tested were: other oral anti-diabetic agents , gender, age (years), race (white/other), glycosylated hemoglobin level , insulin use (yes/no), body mass index BMI; kg/m, alcohol cancers , we alsoThe general characteristics of the study population are described in Table The number of cancer patients in this population was 126 (12.6%). A total of 26% of the subjects used a TZD, including 13.6% on pioglitazone and 12.8% on rosiglitazone , BMI (kg/m2), cigarette smoking, high comorbidity conditions, and number of prescription medications. This model showed a significant association between cancer and the use of any TZD , P = 0.04) (Table P = 0.02) (Table P = 0.76)  (Table P = 0.63) , 1) Table , but not3) Table .P = 0.02) (Table P = 0.001) and number of prescription medications,P = 0.03) (Table P < 0.001) and number of prescription medications,P = 0.02) were associated with cancer history  Table . Other v3) Table . Similarry Table .We have demonstrated an association between the use of TZDs and cancer in a community-based population of adults with diabetes. The association was observed primarily among rosiglitazone users and not among subjects using pioglitazone. Furthermore, the association was found in women, but not men.The gender-dependent observation is difficult to explain. TZDs have gained widespread use in polycystic ovary syndrome (PCOS)  that hasin vitro and our results. Some studies suggest that natural mutant PPARγ alleles might impair native PPARγ function [in vitro studies.Likewise, we are not aware of a convincing explanation or previous results to support the finding in this study of an association with cancer for rosiglitazone, but not pioglitazone. As both are thought to activate PPARγ with similar potency, this finding suggests another mechanism for the association. Furthermore, we cannot explain the differences found between the anti-cancer effect of PPARγ activation function  and diffCancer patients used slightly more prescription medications than non-cancer patients, but similar numbers of anti-diabetic medications Table . This isWe note that cancer patients tended to have more comorbid conditions than non-cancer patients. This could be due to the well-known associations between some of the specific measured comorbidities and various cancers. For instance, chronic obstructive lung disease and cirrhosis are often associated with lung cancer and hepatoma, respectively ,33.To further address potential confounding, we also included two other common oral anti-diabetic agents (sulfonylureas and biguanides). Results presented here showed no significant association of either class of medications with cancer prevalence or any confounding of the association between TZDs and cancer in the general model Table . HoweverThis study has several strengths. First, the interviewed subjects were a randomly selected subset of a large population of patients receiving care in the Northeast USA, and are therefore likely to be representative of primary care patients. Second, all data on medications were obtained by direct observation in the patient's home and not from secondary sources.This study does however also have several limitations, including lack of confirmation of the cancer diagnoses, and lack of information on the time relation between onset of cancer and the use of TZDs or the duration of treatment. In addition, information on the type or stage of the cancer cases is missing, and skin cancers are not reported. As in any cross-sectional study, unmeasured confounders could be responsible for the apparent associations found.Our findings suggest a possible association between cancer and the use of TZDs, particularly rosiglitazone, and particularly among women, but this association requires further investigation.The author(s) declare that they have no competing interests.All authors contributed equally to this workThe pre-publication history for this paper can be accessed here:


Objective. To develop and validate a risk score for detecting cases of undiagnosed diabetes in a resource-constrained country. Methods. Two population-based studies in Peruvian population aged ≥35 years were used in the analysis: the ENINBSC survey  and the CRONICAS Cohort Study . Fasting plasma glucose ≥7.0 mmol/L was used to diagnose diabetes in both studies. Coefficients for risk score were derived from the ENINBSC data and then the performance was validated using both baseline and follow-up data of the CRONICAS Cohort Study. Results. The prevalence of undiagnosed diabetes was 2.0% in the ENINBSC survey and 2.9% in the CRONICAS Cohort Study. Predictors of undiagnosed diabetes were age, diabetes in first-degree relatives, and waist circumference. Score values ranged from 0 to 4, with an optimal cutoff ≥2 and had a moderate performance when applied in the CRONICAS baseline data . When predicting incident cases, the AUC was 0.66 (95% CI: 0.61–0.71), with a sensitivity of 69% and specificity of 59%. Conclusions. A simple nonblood based risk score based on age, diabetes in first-degree relatives, and waist circumference can be used as a simple screening tool for undiagnosed and incident cases of diabetes in Peru. As of 2014, the worldwide prevalence of type 2 diabetes mellitus (T2DM) was estimated to be 9% among adults aged ≥18 years with great impact on mortality, particularly in low- and middle-income countries (LMIC) , 4, untiAn important strategy to prevent or delay T2DM complications is the early identification of those with undiagnosed diabetes; yet, universal screening for diabetes at the population level is not practical in resource-limited settings. The American Diabetes Association recommends the use of glucose test as T2DM screening in people with overweight and obesity as well as in those with other risk factors . As a reAs the American Diabetes Association, the Peruvian Ministry of Health recommends diabetes screening in general population with fasting glucose in adults aged 40 to 70 years with risk factors. However, fasting glucose is not always available in primary care settings, especially in semiurban and rural areas. As a result, a major challenge to be overcome in many countries is the implementation of a simple, fast, and laboratory-free based screening method.Consequently, we aimed to develop a simple laboratory-free risk score to identify people with undiagnosed diabetes and incident diabetes in Peru, a Latin American country that spans coastal, Andean, and rainforest settings. In order to do so, this work benefited from two large-scale population-based surveys: the first one, representative at the national level, was used to develop the score, and the second one, a cohort study, was utilized for external validation. National Survey of Nutritional and Biochemical Indicators for Noncommunicable Diseases (ENINBSC in Spanish), conducted by the Peruvian National Institute of Health [Two different population-based studies were used in this analysis. Thef Health , was usef Health , whose bThe ENINBSC is a national population-based survey carried out in Peru between August 2004 and April 2005, designed to estimate the prevalence of hypertension, type 2 diabetes mellitus, and other risk factors for noncommunicable diseases at the national and regional level . PotentiThe CRONICAS Cohort Study is an ongoing cardiopulmonary project aimed to estimate the prevalence and incidence of hypertension, diabetes mellitus, and obesity in four different settings in Peru that differ in terms of their urbanicity and altitude: Pampas de San Juan de Miraflores, in the highly urbanized Lima, Puno in the altitude  contributing with rural and urban areas, and Tumbes, a semiurban area in the northern coast of Peru . The stuThe procedures of the ENINBSC have been described previously . BrieflySimilarly, the procedures of the CRONICAS study has been published elsewhere . In brieIn both studies, diabetes was defined as any of the following conditions: fasting glucose ≥7.0 mmol/L (≥126 mg/dL) and/or self-report of physician diagnosis. Fasting glucose was assessed by an enzymatic colorimetric method (glucose oxidase GOD-PAP) in both studies. After excluding individuals without known diabetes, undiagnosed diabetes was also estimated to develop and validate the risk score .2), waist circumference , waist-to-height ratio  [Variables included in the analyses were built to guarantee similarities between both studies: sex; age (<55 and ≥55 years); education (in years); self-reported smoking (current versus never/former smoker); alcohol use (user versus never user); self-reported diabetes in first-degree relatives (participant's parents and/or siblings); and levels of physical activity . Anthropometric measurements included in the analysis were body mass index ((BMI), <25, 25–29.9, and ≥30 Kg/md ≥0.70) , and hypd ≥0.70) .http://dx.doi.org/10.1155/2016/8790235).A total of 4,206 participants were enrolled in the ENINBSC, but only 2,472 were included in this analysis. Reasons for exclusion were 1,524 because of age <35 years to make both databases comparable, 129 because of no data about fasting plasma glucose levels being available, and 81 because of known diagnosis of diabetes. In the CRONICAS study, 3,601 participants were enrolled at baseline but only 2,948 records were analyzed as 465 had no data about glucose levels, and 188 were excluded because of previous diagnosis of diabetes. In addition, only data from 2,577 participants was used in the longitudinal assessment of the risk score  with numerical variables. Then, the prevalence and 95% confidence intervals (95% CI) of total diabetes and undiagnosed diabetes were estimated in each study. After that, all cases of known diabetes were excluded from subsequent analyses.p value <0.10 in the bivariate analysis were included in a multiple logistic regression model using stepwise backward elimination with a significance level of 5%. The Hosmer-Lemeshow goodness-of-fit test was used to assess how well the predicted prevalence matched the observed prevalence of undiagnosed diabetes  [The risk score was derived from data of the ENINBSC survey taking into account the multistage sampling strategy of the study. Each potential risk factor  was assessed in bivariate models using logistic regression and undiagnosed diabetes as the dependent variable. Then, risk factors with a ts well) . As we sts well) .For the evaluation of the risk score, the area under the receiver operating characteristic (ROC) curve, sensitivity, specificity, and positive and negative predictive values (PPV and NPV) were calculated. The optimal cut-point was determined using the Youden index, a single statistic that captures the performance of a diagnostic test  . As one We assessed the performance of our score using bootstrap techniques as well as carrying out an external validation using the CRONICAS Cohort Study. Bootstrapping was utilized to estimate confidence intervals for the AUC in our study population. A total of 1,000 random samples with replacement were taken from the development database. The resulting 1,000 prediction models were then assessed to estimate the bootstrap AUC using the bias-corrected version of the confidence intervals . In addiTo evaluate the performance of our algorithm, the Peruvian risk score was compared to previously published models for undiagnosed diabetes including the Brazilian risk score , the Qin Instituto Nacional de Salud and the Centro Nacional de Alimentación y Nutrición, both part of the Ministry of Health in Lima, Peru. In the case of the CRONICAS Cohort Study, protocol and consent forms were reviewed and approved by the institutional review boards of the Universidad Peruana Cayetano Heredia and the NGO Asociación Benéfica PRISMA in Lima, Peru, and the Johns Hopkins University in Baltimore, USA.The protocol and informed consent forms of the ENINBSC study were reviewed and approved by theThe characteristics of participants in both studies are detailed in The overall prevalence of diabetes was 5.1%  in the ENINBSC survey and 8.7%  in the CRONICAS Cohort Study's baseline. After excluding those with known diabetes, undiagnosed diabetes was present in 2.0%  in the ENINBSC survey and in 2.9%  in the CRONICAS Cohort Study.p = 0.21). The Peruvian diabetes risk score was constructed based on the coefficients of that final regression model. The score gave an AUC of 0.73 (95% CI: 0.65–0.78), and the optimal cut-point for undiagnosed diabetes using the Youden index was ≥2 . In addition, when the risk score was evaluated by applying the score to the CRONICAS Cohort Study's population, the AUC for undiagnosed diabetes was 0.68 (95% CI: 0.62–0.73). At the suggested cut-point of ≥2, 42% would be categorized as undiagnosed diabetes with sensitivity, specificity, PPV, and NPV of 70.2%, 58.9%, 4.8%, and 98.5%, respectively . On the p < 0.001), Indian score (p < 0.001), and Qingdao score (p < 0.01) was poorer than our score; however, our algorithm performed similar to the other assessed models, such as the Brazilian risk score (p = 0.93), the Kuwaiti score (p = 0.26), and the patient self-assessment score (p = 0.74), but having only three variables.When previous published algorithms for undiagnosed diabetes were applied to the CRONICAS Cohort Study, the performance of the Rotterdam score  cases per 100 person-years of risk. The AUC of the score was 0.66 (95% CI: 0.61–0.71). With a cut-point ≥2, 42.5% of participants were categorized as at high risk of developing diabetes: sensitivity, specificity, PPV, and NPV were 69.4%, 58.9%, 7.8%, and 97.4%, whereas, for a cut-point ≥1, the respective values were 79.9%, 91.9%, 20.7%, 5.5%, and 98.1%.Using a national population-based survey, a simple nonblood based risk score based on age, history of diabetes in first-degree relatives, and waist circumference was built and shown to perform moderately in detecting undiagnosed diabetes when externally validated. Moreover, the performance of the score was almost similar for detecting incident cases of diabetes in the Peruvian population.A relatively recent systematic literature search found 23 different blood-free prevalent diabetes risk scores: ten from Europe, nine for Asian populations, two from the United States, and two from Middle East . In addiFrom a cross-sectional point of view, with a cut-point ≥2, from 1000 participants assessed by the Peruvian diabetes risk score, a total of 420 would be classified as undiagnosed diabetes with the detection of 20 cases and only 6 will be missing. On the other hand, with a cut-point ≥1, from 1000 screened individuals, a total of 804 would be categorized as having undiagnosed diabetes with the detection of 27 cases and only 7 will be missing. Thus, the reduction of the cut-point of the risk score would increase sensitivity but reducing the specificity and imposing the need of performing a confirmatory test  to almost the double of individuals, with the benefit of having only 7 more people diagnosed.Longitudinally, the same risk score would detect an important number of participants at risk of developing diabetes: 43% of screened individuals would be classified at high risk of diabetes, and of them, 8% would develop diabetes in the next 2.5 years. According to a previous study , 17 repoOur algorithm performed better than the Rotterdam, the Indian, and the Qingdao risk scores in our population, which highlights the need of calibration and/or development of a specific score for different ethnic groups before its adoption. As there are ethnic differences in risk factors for diabetes and Peru is considered a multiethnic country, it is necessary to create specific scores or recalibrate existing algorithms before applying in specific contexts. In addition, with only three variables included, the performance of our predictive model was similar to the other assessed scores included in the analyses. Taken together, the score developed has the potential to augment, in a pragmatic manner, initial rapid screening for diabetes, especially at various nonspecialized primary healthcare services.Our findings also demonstrate that approximately 35% of cases of T2DM (39% in the ENINBSC survey and 33% in the baseline of the CRONICAS Cohort Study) are not aware of their disease. Results are similar to those reported in previous studies in our context  and in sAs the developed risk score is simple, it does not require a blood test or laboratory services, and it might be easily implemented in clinical practice. Moreover, because our score asks for general information in the form of age and diabetes in first-degree relatives and is complemented by a simple anthropometric measure of waist, there is potential for the score to be self-administered.According to our results, any patient aged 55 years and above and having at least one first-degree relative with T2DM has greater probability of having undiagnosed diabetes but also is at risk of developing diabetes in the future. In addition, a greater central obesity, that is, 100 cm or more, independent of the other terms of the score is alone a good predictor of diabetes as reported in previous studies . Our algRecently, the Peruvian Ministry of Health has published the Guide of Clinical Practice for Diagnosis, Treatment and Control of Diabetes Mellitus in Primary Care  and onlyThe strengths of this study include the use of a national population-based survey, including urban and rural areas across major geographical regions, to develop the Peruvian diabetes risk score, as well as its validation using bootstrap but also an independent longitudinal cohort study. Additionally, it is only based on three variables ensuring its simplicity to be used and implemented. However, the study has also some limitations. First, we have utilized fasting plasma glucose as the gold standard for diagnosing diabetes instead of an oral glucose tolerance test (OGTT). Although the OGTT is more sensitive and specific than the fasting plasma glucose, more cases would have been detected with the overload of glucose; it is rarely performed as part of the routine clinical practice. Second, the CRONICAS Cohort Study did not include information from the Amazon rainforest as did the ENINBSC survey. When a sensitivity analysis was performed excluding individuals from the jungle from ENINBSC data, results were similar to those presented in this manuscript (data not shown). In addition, the score was created using a national survey to be applicable to the entire Peruvian population. Third, some variables were not assessed in our logistic regression model such as dietary intake or history of gestational diabetes as such data was not available. As a result, some caution should be made when our algorithm is compared to other risk scores. Fourth, our model is based on the idea of risk stratification instead of individualisation ; for insThe Peruvian diabetes risk score, built using age, self-reported diabetes in first-degree relatives, and waist circumference, proves to be a simple pragmatic screening tool for undiagnosed and incident cases of diabetes in Peru. This experience in generating such simple, easy-to-use approaches for the identification of T2DM can serve to inform other similar LMIC efforts who are on early stages of diabetes prevention. This tool, due to its simplicity, can facilitate various initiatives oriented to introduce and scale up early preventative and management strategies on a wider scale.As 371 (12.3%) subjects were not re-contacted during follow-up, we have conducted a comparison between participants included and excluded from longitudinal analysis.


Our results indicate that the variations of rainfall and water level may have an impact on the aquatic plants in Taihu Lake. The dry period may be not suitable for the growth of the aquatic plants. All aquatic plants in Taihu Lake were dramatically reduced in the dry period, especially for submerged macrophytes and floating-leaf macrophytes. Our results may be helpful for the aquatic restoration in the future.Based on long-term rainfall measurements (1956–2012), water level records (1956–2006), and aquatic plants field survey data (1960–2014), the relationship between precipitation and aquatic vegetation succession in east Taihu Lake, China, is studied. Neither abrupt changes nor any trends were found in the annual rainfall series in Taihu Lake during the studied period (1956–2012). However, for seasonal variations, statistically significant decreases are found in spring and autumn, while the rainfall in winter exhibits statistically significant increase. No significant trend was obtained in summer. A “dry” period was detected in our studied period (1963/1964~1978/1979). Total annual rainfall was significantly positively correlated to the number of rain-days ( The formation, development, and maintenance of wetland ecosystems are closely related to the water resources , 2. The As a functional group, aquatic plants are one of the most important components in aquatic ecosystems , 10. AquSince the beginning of the industrial era, anthropogenic greenhouse gas emissions (largely driven by economic and population growth) and the increasing greenhouse gas concentrations in the atmosphere have led to various changes in the Earth's climate at an unprecedented rate . AccordiGlobal climate change will directly lead to changes in precipitation and the rainfall imbalance. In general, precipitation will be decreased in the low latitude areas, while it will be increased significantly in the high latitude regions . However2 and the average depth is about 1.9 m. The lake is located in the core of the Yangtze Delta within the lower reaches of the Yangtze River Basin, which is the most developed areas in China. Due to the rapid socioeconomic development, the aquatic ecosystem of Taihu Lake has degraded. The distribution and community structure of aquatic vegetation have clearly been changed. The main aim of this paper is to investigate the relationship between rainfall patterns and aquatic vegetation-related processes based on the point of view of plant ecology, by using the long-term (1956–2012) rainfall records from the weather station in Taihu Lake, Dongshan .Taihu Lake is the third-largest freshwater lake in China, occupying a surface area of 2,425 kmhttp://data.cma.cn/). The Dongshan weather station  is located near east Taihu bay , for each xi, the number ri of xi > xj with j < i is computed. Then, the sum is calculated:E) and variance (Var):k is compared with a standard normal distribution at the required level of significance, which is usually α = 0.05 [k| > 1.96. UFk > 0 indicates an increase and UFk < 0 indicates a decrease.To investigate the possible existence of abrupt changes and trends, two methods are adopted in this paper. The first one is the sequential version of the Mann–Kendall test proposed by Sneyers , which iα = 0.05 , 29. Thexi and also going through the whole procedure above, the UBk statistic is obtained. If curve UFk or UBk passes through the 5% significance level from inside outwards, it is considered that the trend is significant. In addition, if curves UFk and UBk clearly cross each other (not overlap) between the critical values at the 5% level, there is an abrupt change, with the intersection point representing the beginning of that change.By using the retrograde series of The second trend estimate approach was computed for each of the four three-month seasons  using data for the period 1956–2012. Trends were computed using the nonparametric median of pairwise slopes method , with st2) of each type of vegetation .There has been extensive amounts of aquatic vegetation field survey data collected from east Taihu Lake. In the current study, we collected a series of published field survey papers and reports covering aquatic vegetation surveys from 1960 to 2014 –36 . In dry period (1963/1964~1978/1979), the rain-days matched with the rainfall volume quite well. A decrement in rainfall depth usually corresponded with a decrease in number of rain-days. However, for those “normal” periods, annual rainfall and annual rain-days did not show this strong relationship. The rain-days remained low in spite of the high precipitation .The annual average of rain-days at Dongshan station was 94. According to the relationship between the anomalies of annual rainfall and annual rain-days . It is reported that the changes of water level may limit the time for plastic responses of morphological and/or physiological traits in aquatic plants. In addition, if the water level change results in high level of disturbance, it may cause physical damage to the plants , which  damage, , 39. Potamogeton macckianus, used to be widely distributed in the middle and lower Yangtze River. In the Taihu Lake, the most produced vegetation type is the emergent species, including Phragmites australis and Zizania caduciflora. Both species' biomass (g/m2) increased from 1960 to 1997; however, it decreased from 1997 to 2014 , Vallisneria spiralis, and Hydrilla verticillata; in 2002, the dominant submerged macrophytes were Elodea nuttallii, Ceratophyllum demersum, and Potamogeton macckianus. For the floating-leaf species, there was a significant increase from 1960 to 2002 and a decrease from 2002 to 2014 ; however, only 40 species was observed in the most recent (2014) field survey . The ext to 2014 . Similar to 2014 . The spe to 2014 . The domPrevious studies , 41 reveNeither abrupt changes nor any trends were found in the annual rainfall series in Taihu Lake during the studied period (1956–2012). However, for seasonal variations, statistically significant decreases are found in spring and autumn, while the rainfall in winter exhibits statistically significant increase. No significant trend was obtained in summer. A “dry” period was detected in our studied period (1963/1964~1978/1979). The other years of the studied period are classified as “normal” period.r = 0.59) and the water level (r = 0.84). However, the strong correlation between annual rainfall and rain-days only occurred in dry period. In those normal period, it is usually found that the rain-days remained low in spite of the high precipitation. Regarding the water level, the strong correlation exists throughout the studied period.Total annual rainfall was significantly positively correlated to the number of rain-days , water level records (1956–2006), and also aquatic plant survey data from 1960 to 2014. The main findings are as follows:


Research on creative cognition reveals a fundamental disagreement about the nature of creative thought, specifically, whether it is primarily based on automatic, associative (Type-1) or executive, controlled (Type-2) processes. We hypothesized that Type-1 and Type-2 processes make differential contributions to creative production that depend on domain expertise. We tested this hypothesis with jazz pianists whose expertise was indexed by the number of public performances given. Previous fMRI studies of musical improvisation have reported that domain expertise is characterized by deactivation of the right-dorsolateral prefrontal cortex (r-DLPFC), a brain area associated with Type-2 executive processing. We used anodal, cathodal, and sham transcranial direct current stimulation (tDCS) applied over r-DLPFC with the reference electrode on the contralateral mastoid  to modulate the quality of the pianists' performances while they improvised over chords with drum and bass accompaniment. Jazz experts rated each improvisation for creativity, esthetic appeal, and technical proficiency. There was no main effect of anodal or cathodal stimulation on ratings compared to sham; however, a significant interaction between anodal tDCS and expertise emerged such that stimulation benefitted musicians with less experience but hindered those with more experience. We interpret these results as evidence for a dual-process model of creativity in which novices and experts differentially engage Type-1 and Type-2 processes during creative production. The study of improvisation is pertinent to any domain that requires adaptation, problem solving, and innovation. The ability to generate, execute, and evaluate choices in real-time can be seen in a range of scenarios from friends having a conversation, to surgeons operating in an emergency room, to musicians performing in a jazz club. In jazz, as in other domains, creative improvisation is developed through rigorous training and experience over many years. Research has begun to offer insights into the structural and functional neural changes that occur as this expertise is acquired Beaty, .In the present study, we tested a dual-process model for understanding creativity  when the cathode was over left ATL and the anode was over right ATL  with tDCS on trained and untrained pianists' finger dexterity and fine motor control  were free of neurological or psychiatric issues and were not taking any neurological or psychiatric medications. They also met the musical requirements: having improvised in a live jazz setting at least 3 times and having at least 10 years of musical training.Jazz pianists from local collegiate music departments, seminaries, bands, and jazz associations in the Philadelphia, PA region were recruited for this study. Due to the highly specialized nature of the sample population, we pursued subject recruitment for 6 months, stopping after we could find no more musicians who met our criteria for participation. Due to the within-subject design of this study, pianists were required to attend three experimental sessions each of which featured a different stimulation-type . Of the 23 musicians recruited, 4 were not able to complete the study due to scheduling conflicts; 1 participant decided not to complete the study; 1 subject's data were not included due to an apparent, unreported, neurological problem. The remaining jazz pianists  at The University of Pennsylvania in Philadelphia, PA. This study was approved by the University of Pennsylvania's Institutional Review Board. At the beginning of the first session, participants signed informed consent, 2 questionnaires as part of a separate study, a handedness inventory, and a mood survey. Upon completion of these surveys, participants were told that they were taking part in a study to examine the effects of tDCS on jazz improvisation without any mention of creativity or expertise. Each participant was given a brief overview of the tDCS equipment, electrodes, and setup while their heads were measured. Once the measurements were complete, participants were fitted with the tDCS electrodes.Logic Pro 9 v.9.1.8  music software recorded the improvisations, collected MIDI performance data, and provided musicians with a bass and drums audio accompaniment. Accompaniments were created through iReal b for Mac OS X v.2.8 , a practice tool with a full rhythm section for any properly formatted jazz chart , sustain pedal, music stand, studio quality headphones, and a binder containing task instructions and jazz lead sheets  were provided for the improvisation task. The experiment's improvisation and recording setup can be viewed in Figure Musicians were randomly assigned to one of six groups which determined stimulation order in their three sessions. The stimulation order was nearly counterbalanced, except for the sham/cathodal/anodal sequence which had one less subject. Musicians were instructed to sit quietly with eyes open while gazing at a fixation cross during the first 6 min of stimulation. The researcher presented the first lead sheet and reminded participants they were now going to improvise to 6, 16-bar jazz songs. Each song included 4 chord cycles and lasted approximately 2 min. The improvisation audio stimulus began with a 4-click count-in, and there were intervals of 15–20 s between stimuli. Musicians improvised with online tDCS for the first 4 takes and offline tDCS for the last 2 in each session. Only the final two offline takes from each session were rated and used in the subsequent analyses. Cognitive demands during online tDCS can influence offline, post-stimulation performance  recorded during sham (n = 34), anodal (n = 34), and cathodal (n = 34) stimulation were normalized to ensure the piano and accompaniment had the same relative volume levels across all takes . The order of the performances was pseudo-randomized for judging with the constraints that the same musician could not be heard consecutively or more than twice within a single judging block. Judging blocks began with an improvisation from an expert and novice improviser from 1 of the first 4 takes in the sham condition. These ratings were not included in the analysis. They were included as a reference point for raters, so they could familiarize themselves with the range of quality of the performances. To determine expert and novice clip selection, we split the participants into quartiles and randomly selected an improvisation from the top (200 or more live performances) and bottom (15 or less live performances) quartiles. Each judge rated the 102 improvisations and 10 baseline takes in 5 blocks of 22–23 improvisations each; however, only the final two, offline takes from each session were included in the analysis. Judging time for each block was approximately 45 min.Upon completion of data collection, the jazz improvisations (Judges scored improvisations on creativity (CR), technical proficiency (TP), and esthetic appeal (AA) on a 7-point Likert scale according to the Consensual Assessment Technique (CAT) Amabile, . This teM = 3.82, sd = 1.29); difficulty of the improvisation task ; ecological validity of the task ; individual practice routines; and experiences improvising in other genres. Values presented here are on a 5-point Likert scale. In the present report, we focus on age and the expertise variables.The demographic musician questionnaire asked basic questions about participants' musical backgrounds and perceptions of the study improvisation task. This included: age; years of music and jazz training; primary performance genre ; number of gigs; degree of comfort improvising jazz  was used to deliver the stimulation current. Thin, saline-soaked sponges were used to interface the 5 × 5 cm rubber electrodes with the scalp. Electrode placement locations were determined using the International 10–20 System. The target-site electrode was placed on the F4 site, approximately overlaying r-DLPFC  to account for inter-individual variation and inter-item variation  hierarchical regression models  goodness-of-fit measure. Changes in −2LL are distributed as χM = 24.2, sd = 4.0), and participants were predominantly male . Expertise data was collected for years of music training , years of jazz training , and number of live jazz gigs performed . The number of gigs covered a large range that spanned 2 orders of magnitude (3–400) and were skewed (skew = 1.95). The number of live jazz performances is an accurate descriptor of a musician's improvisational experience and expertise , technical proficiency , and esthetic appeal . All scales had high reliability, as an ICC > 0.75 is excellent 0.40 to 0.74 is adequate to good, and <0.40 is poor  measured interrater reliability (IRR) for judges' ratings of CR, AA, and TP. Reliability was calculated such that values were computed for consistency where systematic differences between raters are considered to be irrelevant  = 0.96, p < 0.01], CR and TP , AA and TP . These high correlations between scale types may represent the interconnectedness of these three performance features, such that one is needed to express the others in a technically demanding domain like jazz improvisation. Thus, the individual CR, AA, and TP scale-type ratings were averaged to form a single “quality” rating for each improvisation. For further analyses and mixed-effect regression models, the quality rating composite score across judges and scales was the dependent measure for each improvisation.The 3 scale types had highly significant positive correlations after averaging the four judges' ratings for each improvisation: CR and AA [M = 3.85, sd = 1.33). Quality ratings were approximately normally distributed (skew = 0.04), though displaying less peakedness and shorter tails due to negative kurtosis (kurtosis = −1.06). Scores ranged from 1.58 to 6.33, covering almost the entire range of the 7–point Likert scale. No single improvisation received the top score from all judges on all scales, indicating that scores were not clustered at the top or bottom end of the range, avoiding ceiling or floor effects. Although scale-type was not included in the LME regression analyses due to the extremely high correlations between scores on different scales, ratings on the CR scale  were the highest, followed by TP  and AA .Each musician performed 2 takes x 3 conditions ; an overall quality rating was calculated for each take . Thus, in the sham condition, there was an 0.80 increase in ratings per unit increase in expertise. Anodal and cathodal tDCS compared to sham did not affect the quality of performance for the sample as a whole . Figure Estimate = 0.91, SE = 0.32, p = 0.004), and those musicians with the most experience were hindered . Here, ratings of our least experienced participants improved by almost a point when they had anodal stimulation, and the ratings of our most experienced participants decreased by about half a point when they had anodal stimulation compared to sham. The interaction between cathodal tDCS and expertise trended in the same direction but was not significant . There was also a trend for cathodal stimulation to increase ratings for the least experienced musicians , but experts' ratings were not affected .As expected, expertise significantly increased improvisation ratings in the sham stimulation condition  can lead to over-estimation of effect sizes (Button et al., In addition, our sample of jazz pianists included only a moderate range of age and expertise because recruitment was limited by age (older adults producing different responses to tDCS; Fujiyama et al., In this experiment, as in many tDCS studies, localization of the tDCS current is a concern as the pattern of current flow can influence various cortical regions contingent upon individual differences in the geometry of the sulci and gyri (De Berker et al., Although we examined performance in the real-world musical domain of jazz improvisation, the ecological validity of this study may have been somewhat lessened by the stimuli and setting. The chord sequences were loosely based on 16-measure segments of jazz standards, often shifting keys to make them more novel to the performers. While a melody is typically provided on a jazz lead sheet, we did not include a written melody, or “head,” as we did not want sight-reading skills to interfere with one's ability to improvise. While attempting to limit confounding variables, it is unknown how the omission of melodies may have altered the underlying improvisation processes. Additionally, the computer-generated accompaniment did not respond to musicians and had a static tempo; therefore, the soloist could not have the interactions that they would have had in a live jazz setting Monson, .The present study is a first attempt to explore the effects of tDCS on jazz improvisation, a demanding, ecologically valid form of creative expression. Here, we report that brain stimulation differentially influences the ratings of musicians' improvisations dependent upon their degree of expertise. Anodal stimulation to r-DLPFC significantly increased performance quality for the less-experienced pianists while hindering it for those with the most experience. These results provide evidence supporting a dual-process creativity model in which the recruitment of Type-1 and Type-2 processes differs for experts and non-experts. This provides an insight into the neuroplasticity associated with expertise in musical improvisation which may extend to other domains, both artistic and non-artistic.DR, BE, YK, RH, and JK contributed to the conception and design of this work. DR and BE, collected all data. Data analysis and interpretation was conducted by DR, BE, DM, RH, and JK. DR drafted the article, and DM, RH, YK and JK provided critical revisions of the article. Final approval of the version to be published was given by DR, BE, YK, DM, RH, and JK.This project was funded by Drexel University's ExCITe Center - Seed Project 2013-2253.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.