The same STATs and JAKs are also involved in signaling by functionally different cytokines, growth factors, and hormones. Related to this, IFNγ-activated STAT1 binds to the IFNγ-activated sequence (GAS) element, but so do other STATs that are not involved in IFNγ signaling. Activated JAKs such as JAK2 and TYK2 are also involved in the epigenetics of nucleosome unwrapping for exposure of DNA to transcription. Furthermore, activated JAKs and STATs appear to function coordinately for specific gene activation. These complex events have not been addressed in canonical STAT signaling. Additionally, the function of noncoding enhancer RNAs, including their role in enhancer/promoter interaction is not addressed in the canonical STAT signaling model. In this perspective, we show that JAK/STAT signaling, involving membrane receptors, is essentially a variation of cytoplasmic nuclear receptor signaling. Focusing on IFN signaling, we showed that ligand, IFN receptor, the JAKs, and the STATs all undergo endocytosis and ATP-dependent nuclear translocation to promoters of genes specifically activated by IFNs. We argue here that the vacuolar ATPase (V-ATPase) proton pump probably plays a key role in endosomal membrane crossing by IFNs for receptor cytoplasmic binding. Signaling of nuclear receptors such as those of estrogen and dihydrotestosterone provides templates for making sense of the specificity of gene activation by closely related cytokines, which has implications for lymphocyte phenotypes.A small group of only seven transcription factors known as STATs  are considered to be canonical determinants of specific gene activation for a plethora of ligand/receptor systems. The activation of STATs involves a family of four tyrosine kinases called JAK kinases. JAK1 and JAK2 activate STAT1 in the cytoplasm at the heterodimeric gamma interferon (IFN Our understanding of signaling by cytokines such as the interferons (IFNs) at the level of gene activation is stunningly deficient in mechanisms when compared to that of nuclear receptor signaling, as seen, for example, in the case of steroids and their receptors. The canonical model of type I and type II IFN signaling is a representative in fundamentals to that of cytokine or hormone signaling by any protein or peptide signaling via the JAK/STAT pathway.γ (type II IFN) signaling involves basically heterodimeric receptors IFNGR1 and IFNGR2, Janus kinases JAK1 and JAK2, and transcription factor signal transducer and activator of transcription 1α , causing autophosphorylation (activation) of and binding of the JAKs to IFNGR1. Somewhere in the activation process, JAK2 moves from IFNGR2 to IFNGR1 by some unknown mechanism. Also, somewhere in the process, IFNGR1 becomes phosphorylated in the cytoplasmic domain. These events cause binding, phosphorylation, and asymmetric dimer formation of STAT1α. The activated STAT1α, via an intrinsic nuclear localization sequence (NLS), undergoes energy-dependent nuclear translocation to promoters associated with genes that are activated by IFNγ.According to this model, IFNiewed in ). IFNγ bγ, except that there are over 16 different type I IFN subtypes, all of which bind to the same heterodimeric IFNAR1 and IFNAR2 receptor complex. They all use JAK1 and TYK2 tyrosine kinases to phosphorylate STAT1 and STAT2, which in the activated state form a heterodimer, but some are toxic (apoptotic) at high doses while others are not. The toxic type I IFNs such as human IFNα2 bind the receptor with a 10-fold higher binding affinity than does the nontoxic bovine IFNτ, yet they have the same specific antiviral activity on the same cells [α for IFNγ, ISGF3 undergoes nuclear translocation, presumably via an intrinsic NLS. Also, according to the canonical model, ISGF3 is responsible for the specific activation of genes specific for type I IFNs. Thus, signal transduction via JAK/STAT does not explain the unique biological activities of different type I IFNs.Type I IFN signaling is quite similar to that of IFNme cells . There ime cells . Similarλs) further illuminates the inadequacy of the canonical model of JAK/STAT signaling in explaining the mechanism of the specificity of cytokine signaling. Unlike type I IFNs, interleukin 10 receptor 2 (IL10R2) and IFNλ receptor (IFNλR) form the heterodimeric receptor for IFNλ [λs use JAK1 and TYK2 kinases to activate STAT1 and STAT2 for signal transduction. Although the type I IFN receptor is ubiquitous on cells, the type III receptor is cell specific, appearing in particular on epithelial cells and some other cells such as neutrophils. The induction of reactive oxygen species (ROS) in neutrophils is inhibited by IL28A type III IFN, but not by the IFNβ type I IFN [A comparison of type I IFN signaling with that of type III IFNs  and activated STAT1α (pSTAT1α) [α (HP1α) from histone H3, resulting in transcription of genes repressed by HP1α. Activation of JAKs in the nucleus is an important epigenetic event not considered in the canonical model, but is addressed in the noncanonical model of IFN signaling later. We will first discuss briefly the nuclear receptor signaling as it is a big indicator and a guidepost for understanding cytokine signaling as well as that of other peptide/protein signaling in terms of nuclear kinases as well as the role of enhancers/enhancer noncoding RNAs (eRNAs) in such signaling.The recent demonstration of activated JAK2 in the nucleus of cells by gain-of-function mutation (JAK2V617F) or by wild-type JAK2 activated by cytokines or growth factors provides profound insight into the mechanism of cytokine signaling . It alsopSTAT1α) . The earpSTAT1α) . The actγ and its receptor. A comparison of the two at the level of the promoter and enhancer region of genes that are activated by steroids versus those activated by IFNγ readily illustrates the difference in knowledge. In canonical signaling, IFNγ binds to its heterodimeric receptor subunits IFNGR1 and IFNGR2, with IFNGR1 playing a dominant role in the binding [α, which forms an asymmetric homodimer and undergoes active nuclear transport to the promoters of genes that are activated by IFNγ. The canonical model does not provide insight into the movement of JAK2 from IFNGR2 to IFNGR1. Importantly, it does not show the connection between activated STAT1α at promoters of genes that are activated by IFNγ and specific IFNγ function as many cytokines with their own unique functions also activate STAT1α [γ above is not addressed [α in terms of their coordinated function as activated JAK2 in the nucleus is not considered in the NGS studies.As suggested, we do not have comparable understanding of mechanisms of specific gene activation between nuclear receptor systems such as steroid/steroid receptor and canonical JAK/STAT signaling as exemplified by IFN binding . JAK1 ise STAT1α . Next-geddressed . NGS alsγ not only at promoters, but also at enhancers and eRNAs.We have previously presented an overview of nuclear receptor signaling as per steroid hormone (SH)/steroid receptor (SR) signaling . We brieγ signaling by contrast, with only STAT at the promoter, tells us very little and is even primitive relative to that of SH/SR signaling.In a nutshell, SH/SR signaling proceeds as follows. There are many reviews on the subject with considerable detail, but this overview makes our point . SH bindγ signaling but also in general with peptide and protein ligands of hormone, growth factor, and cytokine signaling. The role of enhancers in gene activation is vague and even unknown for almost all ligand/receptor systems except for the case of NRs. Most of the transcribed RNA in the genome does not result in protein production, but rather plays a regulatory role in gene regulation. Of particular interest in the context of enhancers is a subgroup of long-noncoding RNAs (lncRNAs) called enhancer RNAs (eRNAs) [The abundance of genetic and epigenetic mechanisms in SH/SR signaling as well as in other NR signaling at the promoters of genes activated by NRs provides a picture that contrasts not only with IFN (eRNAs) –20. eRNA (eRNAs) , 22.Chromatin immunoprecipitation (ChIP) has been applied to NGS procedures such as ChIP-seq to demonstrate that the estrogen receptor (ER) binds to 5,000 to 10,000 locations across the genome . AnotherActive enhancers are known by the company they keep. Thus, high levels of histone 3 lysine 4 monomethylation (H3K4me), low levels of H3K4 trimethylation (H3K4me3), and increased H3K27 acetylation (H3K27Ac) are associated with active enhancers . These tβ-estradiol (E2) results in E2-bound estrogen receptor α (ERα) association with enhancers adjacent to genes that are upregulated by E2 [α is thus bound to both the enhancers and promoters of genes that are activated by ERα. Similar to the ERα results, studies with dihydrotestosterone- (DHT-) treated prostate cancer cells also showed AR association with an enhancer involved in AR activation of specific genes [In the case of human breast cancer cells, treatment with 17ed by E2 . E2/ERα ic genes . ChIP asic genes . Furtheric genes . The intA current picture of the players at the enhancers and at the promoters of genes activated by steroid hormones such as estrogen and testosterone is presented in γ, details of IFNγ interaction with the IFNGR1 receptor subunit were obtained using X-ray crystallography [γ binding to receptors in intact cells. The structural studies focused on interaction between IFNγ and IFNGR1 extracellular domain via well-defined secondary structures. The C-terminus of IFNγ, which contains a polycationic tail, did not form a clearly defined secondary structure and did not show interaction with the IFNGR1 extracellular domain. Interestingly, the C-terminus polycationic tail functions as a classic nuclear localization sequence (NLS) [γ function [Structural studies of protein/peptide ligands binding to the membrane receptor extracellular domain are generally looked upon as key to gaining mechanistic insight to signaling that occurs at the cytoplasmic domain of the receptor. In the case of IFNlography . IFNGR1 ce (NLS) . To datefunction .γ for the IFNγ receptor in intact cells, IFNγ N-terminus peptide IFNγ (1-39), but not the NLS-containing C-terminus peptide IFNγ (95–132), inhibited IFNγ binding to the receptor on cells [γ. We next used a full-length IFNGR1 soluble receptor and carried out bindings with IFNγ and with overlapping peptides, including IFNγ (1-39) and IFNγ (95–132). We also used overlapping IFNGR1 extracellular and cytoplasmic domain peptides. We discovered that the N-terminus peptide of IFNγ bound to the IFNGR1 extracellular domain and that the C-terminus peptide bound to the IFNGR1 cytoplasmic domain [γ (95–132), as well as the human counterpart peptide, bound to the IFNGR1 cytoplasmic domain region (253–287), adjacent to the binding site of activated JAK2. The binding was specifically blocked by anti-(253–287)-specific antibodies in fixed, permeabilized cells. Related to this, the binding of Sepharose-coupled JAK2 to labeled soluble IFNGR1 was enhanced by IFNγ and IFNγ (95–132), but not by the N-terminus IFNγ (1-39) peptide. Binding was specifically blocked by IFNGR1-binding site peptide IFNGR1(253-287). Such enhanced binding could explain the movement of JAK2 from receptor subunit IFNGR2 to the receptor subunit IFNGR1.In our early studies of binding sites on IFNon cells . This wac domain . Specifiγ and IFNγ (95–132) in treated cells. The first step was to show that internalized IFNγ (95–132) possessed IFN activity. Thus, macrophages with active pinocytosis internalized IFNγ (95–132) and induced antiviral activity as well as upregulation of MHC class II antigens [γ (95–132), Pal-IFNγ (95–132), for internalization by fibroblasts, similarly resulted in the induction of antiviral activity and upregulation of MHC class II antigens [γ peptide Pal-IFNγ (95–132) function which is evidence that the C-terminus peptide functions through IFNGR1.The challenge was to show functional and physical evidence of intracellular IFNantigens . Attachmantigens . Knockouγ also possesses biological activity across species. For example, the following have been reported: (1) liposome-encapsulated human IFNγ induced an antitumor effect in murine macrophages [γ induced an antitumor effect in murine fibroblast cells [γ microinjected into murine macrophages induced MHC class II antigen expression in murine macrophages [γ are odd for two reasons. First, protein ligands like IFNγ are supposed to function by binding to extracellular receptor domains [γ does not have a biological effect on murine cells when simply added to these cells in cultures, because the murine IFNγ receptor extracellular domain does not recognize human IFNγ [γ have had to sit in limbo, waiting for a mechanism of IFN signaling that did not exist at the time of these discoveries.There has long been evidence that internalized IFNrophages , 2) intst cells , and (3)rophages . These ieported:  liposome intst ceman IFNγ . These cγ ligand and IFNGR1 receptor subunit from the plasma membrane to the nucleus. Specifically, we showed that endocytosed IFNγ associates with the cytoplasmic domain of the receptor IFNGR1 subunit in the following manner [γ but not IFNGR1(253-287) intracellular binding site peptide blocked binding of 125I-IFNγ to the IFNGR1 extracellular domain. Internalized IFNGR1(253-287), however, blocked intracellular cytoplasmic binding of 125I-IFNγ to IFNGR1 subsequent to extracellular binding. In the determination of internalization dynamics of IFNγ receptors, we showed that the presence of IFNGR1 and IFNGR2 in the lipid microdomain on the surface of the cell was central to the endocytic events that are linked to the IFNγ noncanonical signaling pathway [γ, receptor subunit IFNGR2 remains in the plasma membrane [γ or if cross-linking in fact occurs in a IFNGR1/IFNGR2 manner [γ attached is exposed to the cytoplasm in endocytic vesicles, since the microinjection of antibodies to IFNγ C-terminus in cells blocked IFNGR1 nuclear translocation as well as STAT1α activation [α activation of STAT1α.Considerable insight has been gleaned concerning trafficking of the IFNg manner . Unlabel pathway . In humamembrane , 41. Thi2 manner , 43. Thetivation . The antγ from the IFNGR1 extracellular domain in the lumen of the endosome and subsequent association with the IFNGR1 cytoplasmic domain in the cytosol of the cell. The mechanism of these events are illustrated in γ receptors present in lipid microdomains that are key to our noncanonical model of signal transduction [+-ATPases or V-ATPases . Theiewed in . Thus, Viewed in . This reγ/IFNGR1 , 49. Thend above , and the 253-287 . The expactivity , 51. Thuactivity  and FiguReceptor tyrosine kinases (RTKs), such as epidermal growth factor receptor (EGFR), have also been shown by different laboratories to undergo energy-dependent nuclear translocation , 53. In γ noncanonical signaling and that of NRs and their ligands. The combination of immunoprecipitation with Western blotting, ChIP assay followed by PCR, nuclear confocal immunofluorescence, and other focused techniques showed that IFNγ, STATα, JAK1, and JAK2 were all present at the GAS element of genes activated by IFNγ [γ NLS in translocation of IFNGR1 into the nucleus. Thus, treatment of cells with IFNγ or with the internalizable C-terminus peptide Pal-IFNγ (95-132) resulted in IFNGR1 translocation to the nucleus with the NLS of IFNγ playing a key role for importin α/β and nuclear pore complex recognition [α, pSTAT1α, and activated JAKs, pJAK1 and pJAK2, form a complex with IFNγ and IFNGR1 at or in the vicinity of promoters of genes that are activated by IFNγ [The data presented here show a remarkable similarity between IFN by IFNγ , 44, 60.-IFNγ 95-2 resulteγ. GAS-luciferase reporter gene transfection, along with IFNGR1 and nonsecreted IFNγ, resulted in enhanced reporter activity. Further, fusion of IFNGR1 to yeast GAL-4 DNA-binding domain resulted in enhanced transcription from the GAL4 response element, consistent with a transactivation domain in IFNGR1 [γ/IFNGR1 in specific gene activation by IFNγ. In NR signaling, the factors at the promoter seem also to be present at the enhancer and we feel that IFNγ, IFNGR1, and pJAK2 should be included in specific gene studies as well as in genome-wide studies.There is evidence that IFNGR1 plays a role in gene activation by IFNn IFNGR1 . These rγ, but was also found in the type I IFN system [α under the same protocol conditions as for IFNγ showed IFNα2, receptor subunits IFNAR1 and IFNAR2, and the type I IFN tyrosine kinase TYK2, all at the response element ISRE of the oligoadenylate synthetase (OAS) promoter [β-actin did not show these IFN players in the same cells treated with IFNα2. Similarities between noncanonical IFN signaling and steroid signaling are presented in Figures Within the IFN family, noncanonical signaling is not limited to IFNN system . Cells tpromoter . An unreα from H3. This, in conjunction with other epigenetic effects (see below) on H3, plays a key role in euchromatin activity related to gene expression in the associated cancers [As indicated, it has been shown that both wild-type and gain-of-function mutated JAK2 perform key epigenetic functions in the nucleus . Mutated cancers . It has  cancers . The que cancers .Essentially lost in the JAK2V617F discovery is the fact that wild-type JAK2, as indicated, also phosphorylated H3 at Y41 (H3pY41) with the same epigenetic result . ActivatChIP-seq has recently been used to study the genomic effects of nuclear JAK1 in an autocrine IL-6 and IL-10 activated B-cell lymphoma . SpecifiAn important functional role of the induction of H3pY41 by the kinase activity of JAK2 and TYK of the IFNs, as discussed here, would be evidence of related nucleosome unwrapping, so that the IFN/receptor complexes have access to the DNA. In this regard, it has been shown that JAK2 induction of H3pY41 increased nucleosome unwrapping for access to transcription factor binding by several folds . Lysine Changes in H3K9 have also been shown to be associated with gene activation . For exaα and DHT/AR paint a picture that is visual and communicative. Promoters contain NR, SRCs, and secondary cofactors like p300/CBP HATs, kinases, and Pol II for transcription iewed in , 65, 66.γt/STAT3 at Th17 cell SEs [Genome-wide NGS with the primary focus on STATs and superenhancer (SE) markers, such as p300 HAT and H3K4me1, showed different patterns for T helper1 (Th1) versus Th2 cells . There acell SEs . The potcell SEs .γ treatment of cells results in a dramatic increase in STAT1 at putative enhancers of IL-6 and tumor necrosis factor [It appears that the cytokines may play a major role as inducers and possibly direct participants of signal-dependent transcription at the enhancers that participate in phenotype-specific gene activation. With regard to STATs and Th1 and Th2 cells and the respective roles of STAT4 (Th1) and STAT6 (Th2), recruitment of p300 and other enhancer signals have been attributed to these STATs based on the absence of p300 at enhancers where STATs are deficient . Master All STATs bind to the GAS element, but how they recognize enhancer DNA is not clear. Cytokines that preferentially activate particular STATs may use another STAT or STATs if there is a deficiency of the preferred STAT. For example, STAT3 may substitute for STAT1 or STAT6 may substitute for STAT5 [α from histone H3, resulting in unwrapping of the nucleosome and exposure of the DNA, presumably at both the promoter and enhancer [γ, activated JAK2 (pJAK2) underwent nuclear translocation along with activated STAT1α (pSTAT1α) [α, activated TYK2 (pTYK2) underwent nuclear translocation [γ and receptor subunit IFNGR1 are part of these nuclear events involving pJAK2 and pSTAT1α, analogous to NR signaling, which could help resolve “the STAT specificity paradox” [We are not aware that any of the extensive NGS studies with STATs has differentiated between activated (phosphorylated) or nonactivated (unphosphorylated) STATs at enhancers or at other regions of the genome. Activated STATs have been reported to be associated with euchromatin, while nonactivated STATs were associated with heterochromatin , 69. Eucenhancer , 26. We pSTAT1α) , 70. Simlocation . In bothOne does not need to fully subscribe to our NR-related noncanonical model of IFN signaling in order to have interest in associations of STATs at promoters and enhancers. We do not yet know if STATs bind to eRNAs, but there is a report of a lncRNA that is involved in conventional dendritic cell (cDC) differentiation which associates with and plays a role in STAT3 phosphorylation in the cytoplasm . Nuclearγ has a C-terminus that qualifies as an IDR [It should be noted that CBP/p300 is highly prone to interactions with intrinsically disordered proteins (IDPs) as well as with proteins with intrinsically disordered regions (IDRs) . IDPs ans an IDR . The proα and DHT/AR, eRNA binds to these receptors as well as to complexes of the mediator, CBP/p300, various other transcription factors, and other players at the enhancer and promoter [promoter , 25, 27.α orchestrate the transcription of eRNA at the enhancers. The eRNA appears to be involved in mRNA transcription at the promoter, which also contains E2 and ERα. Other factors comprise the supporting cast. In canonical JAK/STAT signaling, STAT is the counterpart of the NR and we think that herein is the problem with the interpretation of NGS studies with STATs across the genome that has resulted in “the STAT specificity paradox.” For IFNγ signaling, the inclusion of IFNγ, IFNGR1, and activated JAKs in NGS studies in the context of the noncanonical model could potentially help resolve the paradox.The heart and soul of studies of ligand/receptor signaling is elucidation of the mechanism of specific gene activation. For the case of NRs, the foundations are in place with the NR and ligand at promoters and enhancers of genes specifically activated by the ligand. For example, E2 and ER


All participated in the Ewha Birth and Growth Cohort study. We used a mixed model analysis to estimate the longitudinal association of serum UA concentration due to the rs3825017 , but there were no significant differences by sex. The effects of rs3825017 on UA concentration were significant in boys, but not in girls. Boys with the T allele of rs3825017 had higher concentrations than their counterparts regardless of the time of follow-up. The rs16890979 genotypes were not significantly associated with serum UA concentration in either sex.Overall, the tracking coefficient of UA concentrations in children 3 to 9 years of age was 0.31, and was higher in boys than in girls . Regarding individual variance, serum UA concentrations decreased as age increased (SLC22A12 gene was associated with UA concentration in childhood.This study showed that rs3825017 in the  Uric acid (UA) is the major endpoint of purine metabolism in humans , 2. SeruSLC2A9 encoding GLUT9), solute carrier family 22 (organic anion/cation transporter) member 12 , and the ATP-binding cassette transporter subfamily G member 2 (ABCG2) [SLC22A12 and SLC2A9 have been reported to be responsible for hypouricemia [ABCG2 is also known to be a major cause of gout and hyperuricemia that acts by decreasing urate excretion [SLC2A9 gene had significant effects on gout in an Asian population [ABCG2 on gout was also reported in a meta-analysis  [SLC22A12 related to hypouricemia, c.774G > A and c.269G > A were identified in Japanese [It has been suggested that genetic variants markedly contribute to UA concentrations , with an (ABCG2) . The impuricemia . ABCG2 ixcretion . Among t 0.0001) , but the 0.0001) . As variJapanese  subjectsJapanese . In addiJapanese . Except Japanese , dietaryJapanese , and bodJapanese  are alsoSLC2A9 associated with UA concentrations in children [SLC22A12 and rs16890979 genotype of SLC2A9) of UA-related genes on the UA concentration in childhood using repeated-measures UA data derived from the Ewha Birth & Growth Cohort study to determine whether genetic variants influence the UA concentrations during the first 10 years of life. Because there is little data on how the UA concentration is maintained over time, we also estimated the tracking coefficient of the UA concentrations during childhood.Evidence from GWA and epidemiology studies has suggested a number of candidate genes and a recent study from the Viva La Familia Study also reported genetic variants in children . StudiesThis study was conducted as part of the ongoing Ewha Birth & Growth Cohort study, a single-hospital-based prospective cohort that was established in 2001–2006. The cohort was started to investigate the risk and preventive factors that may be associated with growth and disease susceptibility via longitudinal observations. The study enrolled 940 subjects at 24–28 weeks pregnancy who visited Ewha Womans University Mokdong Hospital, Seoul, Korea for prenatal care. We conducted follow-up when their offspring reached the ages of 3, 5, and 7 years and annually thereafter. By 2016, the follow-up of children aged 9 years had been completed. The methodology of the cohort study has been reported elsewhere . In thisWhen visiting a hospital to participate in the follow-up program, we collected a blood sample from an antecubital vein in a vacutainer tube containing ethylenediaminetetraacetic acid (EDTA). All blood sample were obtained after fasting at least 8 h. Serum UA concentrations were measured using the uricase-peroxidase coupled reaction method on a Hitachi Auto-analyzer 7600  in the Clinical Biochemistry Department of the Seegene Medical Foundation, Seoul, Korea. UA measurements during follow-up were performed in the same way.SLC22A12 and rs16890979 in SLC2A9 were screened using the TaqMan fluorogenic 5′ nuclease assay . The final volume of the polymerase chain reaction (PCR) was 5 μL, and each reaction contained 10 ng genomic DNA, 2.5 μL TaqMan Universal PCR Master Mix, and 0.13 μL 20× Assay Mix. The thermal cycle conditions were as follows: 50 °C for 2 min to activate the uracil N-glycosylase and to prevent carry-over contamination, 95 °C for 10 min to activate the DNA polymerase, followed by 45 cycles at 95 °C for 15 s and 60 °C for 1 min. All of the PCR reactions were performed using 384-well plates on a Dual 384-Well GeneAmp PCR System 9700 (ABI). The endpoint fluorescent readings were performed on an ABI PRISM 7900 HT Sequence Detection System (ABI). Duplicate samples and negative controls were included to ensure the accuracy of genotyping, which was performed at DNA Link Inc. in Seoul, Korea. The distribution of the two SNPs in the study subjects satisfied Hardy–Weinberg equilibrium.The genotypes of rs3825017 in 2 (m2). BMI at each follow-up point was considered a covariate. Birth weight was obtained from the hospital birth records.As covariates, we considered BMI and birth weight, based on previous studies , 18. Matt-test. The average UA concentration according to genotype was analyzed using a generalized linear model.Numerical variables are presented as the mean and standard deviation and categorical variables were presented as the number of subjects and percent. Using repeated-measures data for UA concentrations in subjects aged 3, 5, 7, and 9 years, we estimated the intra-class correlation (ICC) as the tracking coefficient through mixed model analysis . There wTo investigate the long-lasting effects of genetic variants on UA concentration, we used mixed model analysis with a random intercept model. Analyses were conducted using a variance component structure based on lower Akaike information criterion (AIC) values for model selection. The genetic variant used a dominant model (CC vs. CT + TT genotypes) to ensure that the groups had sufficient subjects. Maternal education level and birth weight were treated as time-independent covariates. The BMI at each follow-up visit was considered a time-dependent covariate. To assess the effect modifier, we also included the interaction between sex and genotype as a dominant model.p value < 0.05 was considered statistically significant. For the interaction, P values < 0.1 were considered to be significant, based on a previous report [All of the statistical tests were conducted using SAS 9.4  with two-sided tests, and a s report .p > 0.05, χ2 test) and there was no sex difference in UA concentration with follow-up time (P > 0.05). Overall, the tracking coefficient of UA concentration from 3 to 9 years of age was 0.31, and was higher in boys than in girls . While the tracking coefficient of UA concentration from 5 to 9 years of age was higher , these values can interpreted as showing moderate stability.Two SNPs were examined in 620 children. The rs3825017 allele frequencies of CC, CT, and TT were 61.1%, 34.9%, and 4.0%, respectively. The rs16890979 allele frequencies of GG, GA, and AA were 99.0%, 1.0%, and 0.0%, respectively (Table Table P < 0.001). Individual UA concentrations changed at the same rate with age in the children, indicating that there were no significant effects of a random slope in time. The rs3825017 genotypes had significant effects on the changes in UA concentration in the first decade of life in boys only (Table P for interaction = 0.08). The longitudinal effects of rs16890979 were not assessed due to the very low frequency of the minor allele.The longitudinal effects of rs3825017 genotype on individual UA concentration are presented in Table SLC22A12 had a notable effect on UA concentration in the first 10 years of life in boys. Although there was no sex difference in UA concentration, the genetic effects of rs3825017 on concentration differed by sex (Pgenotype × sex = 0.08) during childhood. The tracking coefficient of UA from 3 to 9 years of age showed moderate stability.This study used a longitudinal approach to examine the relationship between two candidate SNPs and UA concentration in a general pediatric population. The rs3825017 genotype of SLC22A12, which encodes urate transporter 1 (URAT1), and SLC2A9, which encodes glucose transporter 9 (GLUT9), are involved in modulating urate reabsorption in the renal tubules. A meta-analysis of GWA studies identified multiple candidate loci related to UA level in European  and East Asian  populations [SLC22A12 or SLC2A9 were initially reported in Japanese patients with renal hypouricemia, although patients from various ethnic groups have subsequently been identified [SLC2A9 through GWA analysis [SLC2A9 was associated with UA concentrations and it accounted for 4.3% of the variance [SLC2A9 rs16890979 showed possible associations with gout [SLC2A9 rs16890979 in an experimental validation study revealed no significant difference in the expression of GLUT9 [Both ulations . Mutatioanalysis . Of genevariance . This asvariance  and a Czvariance , but thevariance , 27. Altvariance , our stuvariance , 0.23 invariance . In addiith gout . Howeverof GLUT9 . FunctioSLC22A12 affected UA concentration in boys, but not in girls. Three other studies have examined rs3825017 [SLC2A9, several studies have suggested sex differences by genotype including rs16890979; the effects were seen more clearly in women than in men [HIST1H2BF-HIST1H4E, NIPAL1, and FAM35A were novel risk loci [In our study, rs3825017 in s3825017 , 31. In n in men . The sexn in men , the difAlthough the pathological mechanism of UA is unclear, Mazzali et al. reported that rats with induced mild hyperuricemia develop mild hypertension and slightly reduced renal function . In addiEven in children, UA had an independent effect on higher blood pressure after controlling for adiposity factors, renal function, and pubertal status . A cohorTo the best of our knowledge, this is the first study to assess the persistent effects of genetic variants on the UA concentration during childhood. Compared with non-carriers, boys with carriers of at least one rs3825017 T allele exhibited elevated UA concentrations. Our results support those of an earlier study conducted in Korean males . AdditioThis study had several weaknesses. First, we assessed the effect of one tag SNP; other genetic variants near URAT1 may also be important. In addition, residual confounding by unmeasured factors may affect UA concentration. Conversely, our results are less likely to be influenced by unhealthy factors such as alcohol consumption and comorbidities compared to adult studies and case-control studies, because this study examined a general pediatric population. We assumed that the genetic variants were dichotomized by a dominant effect of the risk allele due to the low frequency of homozygous carriers of the risk allele. It is necessary to confirm this in a large-scale study and in studies of different ethnic groups. To verify the statistical power, we calculated the required sample size for linear mixed model with random intercept. As a result, 41 subjects per group were required to evaluate the significance of a difference in the UA concentration between the two groups , assuming that the power was 80% with α = 0.05, four repeated measurements, and mean difference − 11.3. Thus, we found that the statistical power of our study was adequate. Finally, there is a limit to generalizing our findings because the evidence is still insufficient.SLC22A12 and rs16890979 in SLC2A9). Therefore, further studies with extended follow-up periods are required to consider other major genetic loci associated with UA concentrations, such as glucokinase regulatory protein (GCKR) and ABCG2 [In summary, we estimated the stability of UA concentration over 6 years beginning in early childhood, and observed moderate stability. A single measurement of UA during childhood may have a limited ability to predict disease in later life. Through a longitudinal approach, we also found that rs3825017 genotype had a notable effect on UA concentration in boys during childhood. Our study considered only two SNPs (rs3825017 in


Giardia duodenalis and Cryptosporidium spp. are common gastrointestinal protozoa in mammals. Many studies have been conducted on the distribution of G. duodenalis and Cryptosporidium spp. genotypes in sheep and cattle. However, in China, information about molecular characterization and genetic analysis of G. duodenalis and Cryptosporidium spp. in goats is limited. In this study, 342 fecal samples from adult goats were collected from 12 farms in Sichuan Province, China. The occurrence of G. duodenalis and Cryptosporidium spp. in adult goats was 14.9% (51/342) and 4.7% (16/342), respectively. All G. duodenalis were identified as assemblage E, with two novel genotypes (assemblages E17 and E18) being detected at the beta-giardin (bg) locus. Based on three loci—beta-giardin (bg), triose phosphate isomerase (tpi), and glutamate dehydrogenase (gdh)—multilocus sequence typing revealed three novel multilocus genotypes (MLGs) of assemblage E ). Small Subunit (SSU) rRNA-based PCR identified two Cryptosporidium species, namely C. xiaoi (11/16) and C. suis (5/16). This study is not only the first to report C. suis infection in adult goats in China but is also the first to use the MLG approach to identify G. duodenalis in adult goats. Giardia duodenalis and Cryptosporidium spp. are two genera of intestinal parasitic protozoa that infect humans and a broad range of animals, including livestock, companion animals, and wildlife [wildlife –3. Infecwildlife . Damage wildlife ,6.Giardia duodenalis and Cryptosporidium spp. are two of the most common identified parasitic protists in ruminants  and G. duodenalis is considered a multispecies complex with at least eight distinct assemblages (A–H). It has been reported that G. duodenalis assemblages A, B, and E are capable of infecting goats [ng goats , and of ng goats –11. The ng goats –16.G. duodenalis has been identified in a wide variety of animals, including cattle , sheep , goats , dogs , cats , pigs , rabbits (assemblages B and E), rodents , non-human primates (NHPs) , and some other wild animals in China [G. duodenalis infection in goats is limited with reports from only four provinces  [Cryptosporidium spp. has also been widely reported in various animals in China, especially for ruminants, such as cattle, sheep, and goats [Cryptosporidium species  have been reported in goats worldwide with C. hominis and C. parvum frequently reported in humans, especially in developing countries including China, which suggests the potential occurrence of zoonotic transmission between goats and humans [C. ubiquitum, C. xiaoi, C. parvum, and C. andersoni have been identified in goats, with C. ubiquitum and C. xiaoi being predominant [Cryptosporidium spp. genotypes that have the potential to infect goats are still not clear.In recent years, in China . Of the ngjiang) ,13,14. Snd goats . Severald humans . Howeverdominant ,14,19,20G. duodenalis and Cryptosporidium spp. in cattle, particularly calves, and only a limited number of reports have focused on goats as the reservoir for these two parasites [G. duodenalis or Cryptosporidium spp. infection in goats is available in Sichuan Province. To gain a better understanding about the genetic diversity and frequency of G. duodenalis and Cryptosporidium species in adult goats, we conducted the first known molecular study on G. duodenalis and Cryptosporidium spp. infection for Sichuan Province, China. Here, we report for the first time the occurrence and genetic characterization of G. duodenalis infection in adult goats in Sichuan Province, China.Recent studies have mostly focused on reservoirs of arasites . Accordiarasites . HoweverThis study was reviewed and approved by the Institutional Animal Care and Use Committee of Sichuan Agricultural University under permit number DYY-S20174604. The field studies did not involve any endangered or protected animal species. Prior to the study, permission was granted by the owners of 12 farms. No other specific permits were required for the described field studies. In this study, all fecal samples were carefully collected cautiously from the rectum of each goat without causing discomfort.http://ngcc.sbsm.gov.cn/).Sichuan Province, located in southwestern China, has a subtropical monsoon climate, where the breeding industry is still in the initial development stage although it has a long history of traditional husbandry. From June to August 2017, a total of 342 fecal samples were collected from goats at 12 different farms in Sichuan Province, China, with a history of animals with diarrhea, including Qingchuan , Beichuan , Songya , Dalin , Linshui , Shuikou , Shuangliu , Jianyang , Mingshan , Fushun , Naxi , and Jingjiu  . The citThe 12 farms are distributed throughout Sichuan Province, except the western region where animal husbandry is relatively undeveloped. Of the 12 farms, six  practiced intensive feeding, while the other six farms are free-range. All goats included in the study were more than 1 year old. Feces were collected directly from the rectum of each goat into a 50-mL centrifuge tube, which was immediately capped, labeled, and placed in a box with ice. Fecal samples were transported to the Sichuan Agricultural University and stored in 2.5% potassium dichromate at 4 °C. All samples were processed within 24 h after collection.® DNA isolation kit  according to the manufacturer`s instructions and stored at −20°C in 100 μL of the solution buffer provided in the kit [Before DNA extraction, feces were washed with distilled water to remove potassium dichromate. DNA was extracted from the feces using a PowerSoil the kit .Giardia spp., DNA samples were subjected to nested PCR amplifications based on the detection of beta-giardin (bg), triose phosphate isomerase (tpi), and glutamate dehydrogenase (gdh) as previously described [Cryptosporidium spp. was performed targeting the small subunit (SSU) rRNA gene [For detection of escribed . Nested RNA gene . The priRNA gene , 24. TheGiardia and Cryptosporidium species/genotypes, sequences were aligned with reference sequences from GenBank using BLAST (http://blast.ncbi.nlm.nih.gov) and Clustal X. For the phylogenetic analysis, sequences at the three loci obtained in this study were concatenated to form one multilocus sequence for each isolate. A neighbor-joining tree was constructed using Mega 5 (http://www.megasoftware.net/) based on the evolutionary 124 distances calculated using the Kimura 2-parameter model with 1000 replications for the bootstrap analysis.The secondary PCR products were sent to Invitrogen  and sequenced in both directions. To determine the species/assemblages of bg loci; E2 (KT92262) and E3 (KT92259) for the tpi loci; and E3 (KF843925) and E4 (KF843926) for the gdh loci.Representative nucleotide sequences were deposited into the GenBank database under the following accession numbers: MG602956–MG602963. GenBank accession numbers of reference sequences are listed as follows: E5 (KY769092) and E8 (KY633465) for the Giardia duodenalis infection in goats has been reported from many countries such as Ghana, Malaysia, India, Greece, Tanzania, Iran, Spain, the Netherlands, and China [G. duodenalis infection in goats is limited, with only three reports available [G. duodenalis infection in adult goats in Sichuan Province, China.nd China ,16,25,26vailable ,13,14. OG. duodenalis, with the positive rate ranging from 5–37.5%  fecal samples from adult goats, which was higher than that observed in Shaanxi (10.8%) [G. duodenalis in kids in Greece (40.4%) [G. duodenalis-infection may be associated with age, which had been previously proved in sheep and cattle [In the present study, goats from 10 of the 12 farms showed presence of  5–37.5% . The hig1 of 324 .9% fecal (10.8%) , or Heil (10.8%) , but low1 of 324 .9% fecal (40.4%)  are signd cattle ,28. HoweG. duodenalis was analyzed by sequencing the bg, tpi, and gdh loci with 51, 37, and 33 sequences were obtained, respectively , with only one isolate (JY13) identified as E8 (2.0%). Two novel subtypes, named E17 (2.0%) and E18 (2.0%), were also detected. Among the 43 tpi sequences, no genetic variation was observed, with assemblage E2 (97.7%) being predominant and only one isolate (BC07) from Beichuan identified as assemblage E3 (2.3%). At the gdh locus, only two known subtype assemblages, E4 (97.0%) and E3 (3.0%), were found among the 33 sequences, of which the majority of the isolates were identified as assemblage E4, except one isolate from Jianyang (JY13) was identified as assemblage E3. Among the bg, tpi, and gdh loci, predominant subtypes of assemblages E at each locus (E5 (94.1%) at the bg locus, E2 (97.7%) at the tpi locus, and E4 (97.0%) at the gdh locus) were also commonly detected in sheep, cattle, yaks, and pigs, demonstrating a potential risk of cross-species transmission of G. duodenalis to different animals [ animals ,29,31,32bg, tpi, and gdh), forming three novel assemblage E MLGs (MLG-E1 (sc), MLG-E2 (sc), and MLG-E3 (sc)) ) . Assemblectively ,33. Phylectively ,31. MoreG. duodenalis infection in adult goats in Sichuan Province, China. This first-time observation requires a more extensive epidemiological investigation in goats of various ages.In short, our study, for the first time, reports the occurrence and genetic characterization of Cryptosporidium spp., with the positive rate ranging from 2.5% to 19.2%  were identified. C. xiaoi was the most predominant genotype detected in three farms, while C. suis was only found in Naxi. Todate, this is the first molecular identification of C. suis infection in adult goats. C. xiaoi was the predominant species found in this study , which is consistent with that reported in previous surveys in China [C. andersoni and C. ubiquitum are predominant species, respectively. Other studies found that C. parvum was the dominant species in goats [C. parvum, C. andersoni, and C. ubiquitum, which were considered zoonotic subtypes, were not detected. The difference in genotype between our survey and previous studies may be related to host age, raising density, geographical ecological conditions, and examination methods [Among the 16 positive samples, two in China ,19. Howein China , which rin goats –36. In t methods , 19,20.G. duodenalis and Cryptosporidium spp. in adult goats in Sichuan Province, China. We found that assemblage E of G. duodenalis was dominant in adult goats in this region. For the first time, we used an MLG approach to identify G. duodenalis in adult goats, and we detect three novel assemblage E MLGs. For Cryptosporidium spp., two Cryptosporidium species (C. xiaoi and C. suis) were observed, which is, notably, the first time C. suis infection has been identified in adult goats. For a better understanding of the epidemiology and genotypes of G. duodenalis and Cryptosporidium spp. in goats, further investigation with a larger sample of goats of different ages is needed.This is the first known report on the occurrence and genetic characterizations of


To evaluate fertility knowledge and awareness among infertile women attending an Indian assisted fertility clinic and their understanding of the menstrual cycle, how age affects fertility and need for assisted fertility treatment.A cross sectional study was conducted including 205 women seeking fertility treatment at an assisted reproductive unit between March 2017 to August 2017. Patients were interviewed with the help of structured questionnaire by a fertility counsellor. The previous studies were reviewed and a questionnaire was made according to our patient profile and sociodemographic characteristics. Knowledge and awareness was stratified according to socioeconomic status (SES).Most women (59%) were aged between 20 to 30 years indicating concern about their fertility and need for evaluation. More than half (63%) women were from the middle socio-economic strata. Knowledge about fertility and reproduction was low: 85% were not aware of the ovulatory period in the menstrual cycle, only 8% considered age more than 35 years as the most significant risk factor for infertility and most were unaware of when to seek treatment for infertility after trying for pregnancy. Less than half of women understood the need for assisted fertility treatment and donor oocytes in advanced age.Most Indian women across different SES are unaware of the effect of age on fertility. Targeted educational interventions are needed to improve knowledge regarding ideal age of fertility, factors affecting fertility potential and fertility options available for sub-fertile couples. Fertility counselling and information should be provided to young people at every contact with health care professionals. Parenthood is a dream of many couples, however they may not plan a pregnancy keeping advancing age and issues pertaining to infertility in mind . Both unA major factor responsible for delayed child bearing and increasing incidence of subfertility is lack of awareness about fertility potential. Decisions about whether, when and how to conceive, should be a matter of individual or couples’ choice. However, an accurate understanding of reproductive facts is essential for informed fertility decision making. Currently awareness about fertility is low worldwide , 5.Until recent years in India, there was a trend of early marriage and having a first child at less than 25 years, but now due to socioeconomic development and greater interest in education, job and financial settlement there is a delay in parenthood. Most of the women now plan their first pregnancy after 30 years of age. As in other developing countries, in India, having children is the social norm while childlessness is socially stigmatized . InfertiAlthough the incidence of infertility is on rise, no study till date has evaluated fertility awareness among Indian women. Determining the level of knowledge and awareness of fertility practices among Indian women has important public health implications. Targeted fertility education and public enlightenment programmes may help in reducing the number of women experiencing age related infertility and also enable timely referral for assisted fertility treatment. The present study was conducted to evaluate knowledge of women attending infertility clinic regarding factors affecting fertility and availability of advanced infertility treatment options.This was a cross sectional study conducted from March 2017 to August 2017 at infertility clinic of All India Institute of Medical Sciences, New Delhi, India.A total of 205 women attending infertility clinic participated in the study. The inclusion criteria were women of age group 21 to 44 years who either directly consulted the infertility clinic or were referred form general OBGYN clinic and were trying for conception for at least 6 months. Women were invited to participate in the study at the initial consultation and those who voluntarily gave written consent were interviewed by a fertility counsellor. The interview consisted of a nine item questionnaire which was designed after reviewing previous papers on fertility awareness and modified according to patient population and level of understanding in an Indian set up –13. The P < 0.05 was considered to be statistically significant.All statistical analysis was carried out using SPSS, IBM version 19 . Descriptive statistics were used for quantitative variables and were expressed as mean, standard deviation or frequencies, and percentages. The percentage of participants answering correctly was stratified according to SES scale. For comparing categorical data, Pearson’s chi square test or Fishers exact test was carried out as appropriate. For all statistical tests The baseline demographic characteristics of the study population are presented in Table When we assessed the response in relation to age related decline in fertility, it was found that most participants (> 85%) were aware that young women are more fertile and it is easier to achieve pregnancy between 20 to 30 years. This awareness was demonstrated across all socioeconomic strata but significantly better in upper and upper middle class. However, only 26% (53/205) participants could correctly identify the critical age threshold of 35 years after which it becomes difficult to achieve pregnancy. On enquiry regarding the ovulatory period, it was found that 85% participants failed to correctly identify the mid cycle as the most likely phase to achieve pregnancy. This knowledge was consistently poor among all SES classes. Almost 97% (193/205) participants associated past intake of combined oral contraceptive pills with infertility. This belief was similarly demonstrated in all classes of participants. Sixty to 80 % of participants were not aware of the duration after which to consult a fertility specialist if they have been trying for pregnancy and are unable to achieve the same. Only 26% women were aware of the need of assisted reproduction and donor oocyte at the age of 50 years. On subgroup analysis according to SES, women in the upper SES had better knowledge (39% correctly answered) regarding need of ART and the option of donor oocyte in advanced age than in other categories. Similarly, knowledge regarding surrogacy was also higher among women belonging to upper SES category than in the lower category (more than 50% in upper SES correctly answered versus 24% in lower SES).There is a worldwide increase in age at first conception and decline in fertility rate . InfertiOur study population consisted of women attending infertility clinic who either consulted directly or were referred from general OBGYN clinic. These women are supposed to have more knowledge about infertility than general population. Although majority of participants (85%) were aware that younger age group (20 to 30 years) is more fertile and fertility declines with age however they were unable to identify the critical threshold of 35 years after which fertility rapidly declines. The participants in the upper and upper middle SES class had better knowledge compared to the lower SES classes. Similar findings were also reported in a study of fertility awareness among oocyte donors in Spain in which the majority of participants (75%) overestimated the age of decline in fertility . AnotherOur study also highlights another important finding regarding association of oral contraceptive pills and infertility. Nearly 97% of women believed that past history of pill intake was associated with infertility. People often incorrectly attribute the age related decline in fertility or other unexplained cause of infertility to contraceptive pill use. In a study conducted in African women between 37 and 62% of women believed that contraceptives could harm the womb . ImplemeThe majority of respondents (74%) in our study believed that it was possible to achieve pregnancy at 50 years either naturally or with the help of ART with self oocytes. This over optimism and belief that it is possible to have children naturally until the onset of menopause has also been shown in previous studies .Knowledge of the need of donor oocytes and assisted reproduction in women of advanced age was better in upper and upper middle SES classes. Patients in the upper SES were significantly more aware about surrogacy and its implications. Bunting et al. have shown in their study that variation in fertility knowledge is mainly related to socio-demographic factors like education, employment and development index . SimilarMost of the studies on fertility awareness have been conducted in women only. Since, men play a major role in childbearing decisions and timing, it is important to assess their level of understanding and knowledge of factors affecting fertility .Taking into account low fertility knowledge among men and women, British Fertility Society has started Fertility Education Initiative (FEI) in association with Royal College of Obstetricians and Gynaecologists and few other stakeholders to help all women and men so that they are able to make informed choices about their fertility planning . The FEITaking into account the increasing prevalence of infertility and referrals to ART clinics, accurate fertility information should be provided widely to the masses through awareness programmes, media and at the time of contact with healthcare professionals. Programmes similar to FEI can be started in India which can be integrated with education curriculum to increase fertility awareness. As it is with other areas of health, one should be empowered with sufficient knowledge about fertility potential and planning. It is mandatory to have methods and tools available to spread fertility awareness among all potential couples, college students and carrier settlers so that they will not face the reality of infertility in the twenty first century.Our study is the first of its kind to be conducted in India and has highlighted some very important issues of both national and international significance. We assessed the SES of the study population which has not only provided a better understanding of variation in knowledge among different SES categories but also indicated that higher socioeconomic status does not necessarily mean a better awareness of fertility knowledge. However, there are some biases in our study. We interviewed women from the fertility clinic of a tertiary care centre and they may not represent the general population. Prior consultation for infertility has been found to improve knowledge in some areas of infertility .The present study has identified significant important gaps in women’s knowledge and awareness regarding fertility practices. These include poor knowledge of factors affecting fertility especially age related decline in fertility, fertile period in the menstrual cycle and myths regarding oral pill intake and risk of infertility. The study also concluded that higher socioeconomic status and better education does not translate into improved knowledge and awareness of factors affecting fertility. Better accessibility of higher SES classes to media and internet may have enhanced their knowledge regarding advanced fertility treatment options, but they still lacked the basic knowledge of fertility affecting factors. Hence, there is a need of generalized education of masses and targeted interventions at primary level. The most important information that needs to be transmitted to young infertile couples is when it would be biologically too late to achieve childbearing and availability of other ART facilities at late age. Future research should focus on assessing the knowledge of infertile males and also professionals involved in primary health care.


Protein kinases modulate the reversible postmodifications of substrate proteins to their phosphorylated forms as an essential process in regulating intracellular signaling transduction cascades. Moreover, phosphorylation has recently been shown to tightly control the regulatory network of kinases responsible for the induction and maintenance of pluripotency, defined as the particular ability to differentiate pluripotent stem cells (PSCs) into every cell type in the adult body. In particular, emerging evidence indicates that the balance between the self-renewal and differentiation of PSCs is regulated by the small molecules that modulate kinase signaling pathways. Furthermore, new reprogramming technologies have been developed using kinase modulators, which have provided novel insight of the mechanisms underlying the kinase regulatory networks involved in the generation of induced pluripotent stem cells (iPSCs). In this review, we highlight the recent progress made in defining the roles of protein kinase signaling pathways and their small molecule modulators in regulating the pluripotent states, self-renewal, reprogramming process, and lineage differentiation of PSCs. Pluripotent stem cells (PSCs) have unique properties allowing them to undergo unlimited self-renewal and retain pluripotency to differentiate into any cell type in the developing organism, providing a precious source of cells for applications in regenerative medicine , 2. PSCsin vivo and their growth features in vitro [in vitro, similar to that of naïve PSCs, but cannot generate germline-competent chimeras in vivo [β (TGFβ) serine/threonine receptor kinase signals [PSCs are classified into molecularly distinct “naïve” and “primed” pluripotent states based on their potential to develop into the germline lineages in vitro . Primed  in vivo . In addi in vivo . Naïve P in vivo . In cont signals –12.in vivo [in vivo following injection of a single cell [in vitro [Although PSCs can differentiate into all kinds of cell types in an adult organism, they have limitations in their ability to develop into the extraembryonic placental tissue in vivo . Recent gle cell –16. LCDMgle cell . In contin vitro , 16.β serine/threonine receptor kinases signaling [The acquisition and maintenance of pluripotency are intrinsically associated with the core regulatory networks of particular transcription factors such as Oct4, Sox2, and Nanog . Severalignaling  , PI3K, and Erk. Autocrine FGF4-Erk1/2 signaling has an important role in promoting the transition from a naïve to a primed state and stabilizing the primed cell state [The MAPK/Erk signal transduction cascade transduces the environmental signal of growth factors such as FGFs, and the optimal level of Erk signaling is critical for self-renewal and pluripotency . FGFs anll state . Moreovell state . LIF supll state . LIF alsll state . Indeed,ll state .However, the pharmacological inhibition of Erk signaling by PD0325901 was found to be insufficient to maintain mESCs over the long term or clonally without LIF supplementation, implying that there are alternative signaling pathways associated with pluripotency , 28. CheAlthough hESCs show increased STAT3 phosphorylation in response to exogenous LIF, LIF addition does not seem to enhance self-renewal in culture . It is wβ-catenin is a key pathway involved in embryonic development and has proven to support the pluripotency of both mouse and human ESCs [β-catenin, leading to long-term self-renewal primarily by abrogating function of the T-cell factor 3 (Tcf3) transcription factor which acts as a repressor on the pluripotency network, such as Oct4, Sox2, and Nanog [β-catenin target genes, such as brachyury [β-Catenin can also physically interact with Oct4 resulting in the upregulation of Nanog expression in a potential Tcf-independent manner [Wnt signaling via man ESCs . Activatman ESCs , 38. Addman ESCs , 34. GSKnd Nanog , 40, andrachyury . β-Catent manner .β-catenin by tankyrase inhibition, can promote the establishment of homogenous primed-state mEpiSCs [β-catenin complex, and thereby increasing cytoplasmic retention of β-catenin and preventing the β-catenin-TCF interaction [Dual inhibition by CHIR99021 and PD0325901, so-called two inhibitor (2i), can drive self-renewal and inhibit differentiation and conversion of naïve mESCs . In cont mEpiSCs , 43. Sim mEpiSCs . However mEpiSCs  and stab mEpiSCs . It is seraction . In addieraction .The PI3K/AKT/mTOR signaling cascade helps to regulate a variety of cellular processes, including cell proliferation, growth, survival, and metabolism. PI3K/AKT can be activated by insulin/insulin-like growth factor 1 (IGF1) in both mouse and human ESCs. mTOR is a conserved protein kinase that is active in two distinct complexes, mTORC1 and mTORC2. The genome-wide CRISPR-KO screen reveals that receptor tyrosine kinase- (RTK-) mediated AKT activation and the mTORC1-negative regulators such as Tsc1/2 and Gator1 play an important role in the regulation of appropriate GSK3 activity for naïve pluripotency in mESCs. Interestingly, the loss of Tsc1/2 complex causes AKT/mTORC1-dependent GSK3 inhibition, but the loss of Gator1 complex shows the opposite phenotype of GSK upregulation .In both mouse and human ESCs, this PI3K/AKT/mTOR signaling pathway is essential for maintaining pluripotency . Thus, Pβ superfamily of structurally related cytokines, including TGFβ, BMPs, and activin/nodal, plays key roles in regulating the stem cell fate. TGFβ signaling is initiated by the binding of ligand to its serine/threonine kinase receptors and phosphorylation of the cytoplasmic signaling molecules Smad2/3 for the TGFβ/activin/nodal pathway or Smad1/5/8 for the BMP pathway. During embryonic development, epithelial cells undergo a morphogenetic event known as the epithelial-to-mesenchymal transition (EMT), in which the cell polarity and cell-cell adhesion are lost toward differentiation to mesenchymal cells. TGFβ signaling can induce the EMT, which is essential for developmental processes, including mesoderm and neural tube formation [The TGFormation , 53.β signaling is essential in sustaining the pluripotency of hESCs and mEpiSCs, and its modulation can lead to direct lineage-specific differentiation [βRI kinase inhibitor SB431542 leads to a reduction in Oct4 and Nanog expression [β/activin/nodal signaling, but not BMP, is necessary for proliferation of mESCs [TGFntiation . By contntiation , 55. Indpression  and diffpression , althougpression . By contof mESCs .Functional screening and phosphoproteomic profiling suggested novel roles for kinase signaling pathways in modulating the stem cell fate . Cyclin-Human naïve PSCs are a recently derived population from human embryos or primed hPSCs , 63–65. β/activin/nodal pathway) [Moreover, the LIF/2i/PD173074 tyrosine kinase inhibitor promotes the transition of human primed PSCs to naïve PSCs or their maintenance in a naïve pluripotent state, in combination with Oct4, Klf4, and Klf2 , and alspathway) . LIF/3i pathway) . Howeverpathway) . These rβ/activin/nodal pathway D4476 were shown to facilitate the reprogramming of mouse embryonic fibroblasts (MEFs) to iPSCs in the absence of Sox2 and Oct4, respectively [β/activin/BMP pathway including activin-like kinase (ALK) 4/5/7) is also shown to increase the efficiency of the reprogramming of human somatic cells to iPSCs in combination with PD0325901 and the ROCK inhibitor thiazovivin [Small molecule compounds provide a useful supplement in the development of a more efficient and safer method to generate clinical-grade iPSCs , 72. Selectively , 74. Simazovivin . In addiβ signaling pathway plays an important role in EMT, and its inhibition can consequently enhance mouse and human reprogramming [The mesenchymal-to-epithelial transition (MET) is the hallmark crucial event toward the derivation of iPSCs from somatic cells, which is coordinated by repression of the EMT . The TGFgramming . A kinomgramming . A recengramming .κB, and tyrosine kinase pathways) stimulates glycolytic metabolism and enhances the reprogramming of human somatic cells to iPSCs [The metabolic shift from oxidative phosphorylation (OxPhos) to glycolysis is also a crucial event in somatic cellular reprogramming. Both naïve and primed PSCs show increased dependence on glycolysis under aerobic conditions with severe OxPhos suppression , 80. Queto iPSCs , 81. PS4to iPSCs . Collectβ type I receptor kinase (ALK5) inhibitor and a chemical replacer of Sox2, and tranylcypromine, a monoamine oxidase inhibitor) can generate iPSCs from mouse and human somatic cells only with the exogenous Oct4 factor [α agonist), decitabine (a DNMT1 inhibitor), EPZ004777, and SGC0946 , leads to up to a 1000-fold increase in reprogramming efficiency [μM during the early stage for 16–20 days in the chemical reprogramming process not only facilitates the generation of CiPSCs from neonatal and adult fibroblasts [μM CHIR99021 supplemented to AM580 and EPZ004777 promote the formation of XEN-like colonies by 2- to 3-fold [The chemical compounds VC6T  embryonic-like programs consequently accelerate the chemical reprogramming process with HDAC inhibition . Until nβ/activin/nodal signaling as well as enhances the differentiation of neural progenitor cells from hPSCs in combination with the LDN193189 BMP pathway inhibitor [The balance between the self-renewal and differentiation of PSCs is crucial for the developmental process and tissue homeostasis . Specifinhibitor , 95.PSCs have characteristic protein-protein networks that change dynamically during differentiation . A phospThe Src family of nonreceptor tyrosine kinases plays an important role in diverse cellular regulation of adhesion, proliferation, growth, and survival , 98. mESβ family, have been well characterized, the diverse novel kinase networks that modulate self-renewal and pluripotency still need to be identified, which requires more comprehensive phosphoproteomic and functional kinome analyses. It is also necessary to investigate the elaborate molecular mechanisms of distinct pluripotent states requiring kinase functions in PSCs, which will allow for the discovery of new molecular targets linked to the PSC fate. Furthermore, the use of small molecule compounds that modulate kinase activities will also help to unravel the underlying molecular mechanism of pluripotency-associated kinases, while further providing a powerful tool for modulating the PSC fate, thereby contributing to promoting the field of regenerative medicine.This review summarizes the role of pluripotency-associated kinases in regulating the unique characteristics of PSCs with regard to pluripotent states, self-renewal, reprogramming process, and lineage differentiation. Achieving microenvironmental control using specific kinase inhibitors can help to regulate various kinase signaling networks, which can ultimately significantly affect the expression or function of pluripotency and/or reprogramming factors that determine the PSC fate. This integration of molecular mechanisms suggests that pluripotency is largely maintained or induced through posttranslational modifications. Although different signaling pathways, including the FGF/Erk pathway, PI3K/AKT/mTOR signaling, LIF/JAK/STAT3 axis, Wnt/GSK3 signaling, and TGF


Dear Editor,Atypical chronic myeloid leukemia (aCML) is a rare haematological malignancy classified as a myelodysplastic/myeloproliferative neoplasm (MDS/MPN). Diagnostic criteria include leucocytosis with left shift, dysgranulopoiesis, minimal basophila and monocytosis, a hypercellular bone marrow, less than 20 % myeloblasts in the blood or bone marrow, and absence of the distinct molecular abnormalities of classical MPN . While Several studies have investigated the mutation profile of sizeable cohorts of aCML patients in chronic and blast crisis phases by targeted sequencing of recurrently mutated genes of myeloid malignancies. However, in order to comprehensively survey the current mutational landscape of aCML, these studies together with smaller series and case reports adopting such targeted sequencing approaches in the literature, are summarised in Figure 1ASXL1, NRAS, SETBP1, SRSF2 and TET2. In addition to broad cytoreductive and supportive measures , clinical., 2015; Langabeal., 2015).Due to the lack of a standard of care combined with the clinical and genetic heterogeneity of this malignancy, a personalized prognostic and therapeutic direction to aCML management is anticipated, facilitated by the progressively fundamental role of targeted mutation profiling.The author declares no conflict of interest.


Hepatozoon protozoan parasites from domestic cats in South Africa was assessed and compared against published data to assess global biogeographic patterns. Multiple distinct haplotypes of Hepatozoon felis were identified, as well as an unrelated Hepatozoon lineage. Hepatozoon felis genetic diversity globally is very high, indicating a likely complex of species. The recently described Hepatozoon apri from wild boars is closely related to some lineages of H. felis. Sarcocystis and Babesia parasites were also detected. Since Hepatozoon felis is apparently a species complex, potential differences between genetically distinct forms need to be assessed. The finding of an unrelated Hepatozoon indicates that felids can be infected by more species of Hepatozoon than currently known, and that trophic interactions may increase the number of Hepatozoon species found in carnivores. Genetic screening again is demonstrated to identify previously unrecognised parasites from vertebrate hosts.Genetic diversity within partial 18S rRNA sequences from  Hepatozoon are apicomplexan parasites infecting most terrestrial vertebrate groups, having haematophagous arthropods as the definitive hosts. Recent phylogenetic assessments have shown a complex pattern of relationships, with the genus Karyolysus forming a clade within a paraphyletic Hepatozoon, although parasites from most mammalian carnivores form a well-defined genetic lineage . Sequences used are given in Supplementary H. felis-like’ sequences that were included in the estimate of phylogenetic relationships and network with other data from GenBank  and an unknown Hepatozoon sp. of the ‘Bartozoon’ lineage (Karadjian et al. Vulpes pallida, sometimes infected with Hepatozoon typical of desert rodents (Maia et al. Hepatozoon should preferably sequence positive amplifications, rather than assuming that Hepatozoon in felids will correspond to H. felis or H. canis.The finding of another, unrelated type of Sarcocystis species is not unexpected, as these primers have been shown to amplify these parasites before (Harris et al. Sarcocystis are known to use cats as definitive hosts (Levine & Tandros S. nesbitti warrants further attention as this species can cause illness in humans, but is thought to use snakes as the definitive host and humans as occasional intermediate hosts (Lau et al. Sarcocystis from a domestic cat may be related to S. nesbitti.The amplification of a H. felis as currently constituted includes highly distinct genetic lineages, and a detailed assessment with additional and more variable genetic markers is needed to determine how many actual species exist under this name. In the meantime, these lineages can be referred to as the H. felis-like complex. Relationships between these H. silvestris, H. martis and H. apri also need to be elucidated. Domestic cats, as well as being infected by three currently described species of Hepatozoon, H. felis, H. silvestris and H. canis, can also be infected by an unrelated lineage typically observed in rodents and reptiles that may be another example of infection via a trophic pathway. Therefore, the full list of Hepatozoon species that can be found in cats may still be incomplete. Similarly, the finding of an unknown Sarcocystis species from a domestic cat indicates that the list of these species infecting cats also needs clarification. Finally, while some general biogeographic patterns are apparent, many areas, including South Africa, can harbour highly distinct genetic lineages.To conclude,


In the TSO treatment subsection of the Materials and methods section, the concentration of betamethasone valerate ointment is incorrect. The correct sentence is: Patients were applied with 0.12% betamethasone valerate ointment to the lesion twice a day after washing or bathing by parents.


Baduanjin, a traditional Chinese exercise therapy, has been widely used in China to treat type 2 diabetes (T2DM) with depression (DD). However, the underlying mechanism of Baduanjin in anti-DD is unclear. This study was focused on investigating the effects of Baduanjin on symptoms of depression and blood glucose in patients with DD and the underlying mechanism.We performed a 12-week Baduanjin intervention on patients with DD and longitudinally compared the differential expressions of lncRNAs, circRNAs, and mRNAs between pre- (BDD) and post- (ADD) Baduanjin intervention in the same group. Subsequently, Gene Ontology (GO) and pathway analysis was performed to investigate the function of differentially expressed mRNAs. Finally, Reverse Transcription-Polymerase Chain Reaction (RT-PCR) was used to verify the sequencing result and the mRNA-lncRNA regulatory network was constructed.The blood glucose level, depression index scores, and PHQ9 scores of the patients with DD were significantly decreased (P < 0.05) after Baduanjin intervention. Compared to BDD, 207 lncRNAs, 266 circRNAs, and 610 differentially expressed mRNAs were identified in ADD. Kyoto Encyclopedia of Genes and Genomes (KEGG) and GO showed that the significantly dysregulated mRNAs were mainly involved in immune function and inflammatory response pathways, and various signaling pathways including IL-17 and TNF. In addition, we selected five differentially expressed lncRNAs to construct an lncRNA-mRNA regulatory network, and found a total of 1045 mRNAs associated with them.Our research is the first systematic profiling of mRNA, lncRNA, and circRNA in patients of ADD and BDD, and provides valuable insights in the potential mechanism of Baduanjin in anti-DD. Further, it was confirmed that Baduanjin is a safe and effective intervention for patients with DD because it can effectively ameliorate the symptoms of depression and blood glucose levels in patients with DD by regulating the dysregulated expression of lncRNA, mRNA, and circRNA.The online version of this article (10.1186/s13020-019-0225-1) contains supplementary material, which is available to authorized users. Baduanjin (Eight-Section Brocades) is a traditional form of exercise that has been extensively practiced in China for many centuries . BaduanjPeople with T2DM have a 24% higher risk of depression than healthy people, and people developing depression are more likely to develop T2DM , 11. PatAbnormal expression of RNA during transcription is involved in the pathogenesis of DD , 14. LonThe Minimum Standards of Reporting Checklist contains details of the experimental design, and statistics, and resources used in this study  (2016BZHYLL0105) and conducted in accordance with the principles of the Declaration of Helsinki. All enrolled patients were from hospitalized patients at the BUCM affiliated hospital (2016.1–2016.4) and were confirmed to have signed informed consent after enrollment. Cutoff points of 44  and 55 (standard score) were used to define depression for the Self-Rating Depression Scale (SDS), whereas a score of 5 or more was used as a cutoff to define depression for patient health questionnaire 9 (PHQ9). We performed a 12-week Baduanjin intervention on DD patients enrolled in the study (starting at 8:30 am every day for approximately 30 min). Fasting blood samples of patients in the DD group were collected before and after the intervention of Baduanjin for further experiments. More specifically speaking, at 8:00 am on the 0th and 12th week of the intervention, the patient was in a state of peace and fasting, and blood was collected by nurse.Total RNA was extracted from whole blood according to a previously reported method . BrieflySequencing libraries of well-pooled different samples were denatured by 0.1 M NaOH to generate single-stranded DNA, which was in situ expanded on TruSeq SR Cluster Kit v3-cBot-HS  after dilution to 8 pM concentration The resulting fragment ends are sequenced using a Sequencer such as the Illumina HiSeq 4000 for 150 cycles.Transcript abundance calculations were performed by software StringTie by aligning the results to known transcriptomes and then calculated by Ballgown. The units of expression were expressed as FPKM (Fragments Per Kilobase of gene/transcript model per Million mapped fragments). Genes were expressed at a threshold of FPKM ≥ 0.5 and the average number of genes with FPKM exceeding 0.5 in each group was considered to be expressed in the subgroup and statistically analyzed.The expression of circRNA (Backsplice junction reads count) was quantified by the software STAR and then compared to the known transcriptome for junction locus detection and then calculated by CIRCexplorer2 for Backsplice junction reads. Unlike genes, the circRNA expression threshold is that CircRNAs with mean CPM exceeding 100 in each group are considered as expressed in the group and statistically analyzed.−△△CT relative quantification method and normalized to β-actin to calculate relative lncRNA and mRNA concentrations. All primers are shown in Table qRT-PCR was performed to verify the expressions of lncRNAs and mRNAs related to the occurrence and development of DD. Purified total RNAs isolated from 14 samples were reverse-transcribed into cDNA according to the manufacturer’s instructions. Data were analyzed using the 2We used software topGO for Gene Ontology (GO) analysis to obtain significantly enriched GO articles and corresponding genes to deduce important biological functions involved in differentially expressed genes. Meanwhile, based on the newest KEGG database (Kyoto Encyclopedia of Genes and Genomes), find out the pathways which are most associated with differentially expressed genes.GSEA is a computational method used to determine whether a given gene set has significant differences among different groups . Genes iAn lncRNA-mRNA regulatory network was constructed based on the inter-regulatory association between differentially expressed genes in the blood of DD patients following the Baduanjin intervention.P value the greater the contact between the GO and pathway terms and the input differentially expressed genes. Two-sided P values < 0.05 were considered statistically significant.The results are expressed as mean ± standard error (SEM). GO and pathway enrichment analysis was done using Fisher’s test, the smaller the We compared fasting blood glucose levels (GLU), HbA1c %, SDS, and PHQ9 in patients with DD before and after the Baduanjin intervention. The results showed that after the intervention of Baduanjin (ADD), HbA1c % did not significantly differ than that before Baduanjin intervention (BDD). However, the levels of fasting blood glucose, PHQ9, total coarse points, and standard score of ADD were significantly lower than those of BDD Fig. .Fig. 1EfCompared to BDD, 610 (99 up-regulated and 511 down-regulated) and 266 (170 down-regulated and 96 up-regulated) differentially expressed mRNAs and circRNAs were identified in ADD, respectively  to construct the lncRNA-mRNA regulatory network map. A total of 1045 related mRNA were identified, of which 688 were negatively correlated with the corresponding lncRNAs and 357 mRNAs were positively correlated with the corresponding lncRNAs  in BDD group, which is consistent with previous studies [In this study, we found that Baduanjin intervention significantly reduced blood glucose levels, and lowered the depression coefficient in patients with DD, showing an anti-DD effect. Results of the current study are in line with a previous review indicating that physical activity has a positive effect on regulating mood and reducing the symptoms of depression . 610 and studies , 23. It  studies –26.Fig. lncRNA-DLGAP1-AS1 was down-regulated in ADD. This shows that Baduanjin may play a role by down-regulating DLGAP1-AS1.The disks large-associated protein (DLGAP) family serves as a scaffold in postsynaptic density, which can link the glutamate receptors in the postsynaptic membrane with other glutamate receptors, signaling proteins, and cytoskeletal components . With thSince most of the differentially expressed lncRNAs have not been studied, we have constructed mRNA expression profiles to help study the effect of Baduanjin on DD. Depression can be perceived as a psycho-neuro-immunological disorder and the chemokine system plays a crucial role in its pathogenesis –32. In tKEGG pathway analysis revealed that Baduanjin may play a pivotal role in DD progression through the significantly enriched 20 pathways including IL-17 and TNF. Our results support previous discovery , 36. StuThe improvement of the depression index and blood glucose levels in patients with DD by Baduanjin intervention was significantly associated with the significant enrichment of GO processes in the blood of these patients. According to the annotation results, the most significant GO processes are cellular response to lipopolysaccharide, nuclear-transcribed mRNA catabolic process nonsense-mediated decay, and inflammatory response, indicating that the related coding genes contributed to development of DD. Interestingly, the top up-regulated biological processes belong to the immune pathway, which is closely related to depression . TherefoIn addition, five significant differentially expression mRNAs  were identified in the lncRNA-mRNA regulatory network and defined as the key nodes of the network. Among them, differentially expressed mRNA-CXCL8 was positively correlated with AC068580.5, DUSP2 was positively correlated with RP13-516M14.10, mRNA-OSM was positively correlated with RP13-516M14.10, AC068580.5, and CTD-2530H12.2, mRNA-CXCR4 was positively correlated with RP13-516M14.10, CTD-3014M21.1, AC068580.5, and CTD-2530H12.2, and mRNA-NR4A2 was positively correlated with RP13-516M14.10. The lncRNA-mRNA regulatory network analysis confirmed the crosslinking and complex regulation patterns of lncRNA and mRNA expression and verified the authenticity of the regulatory network. DD-related lncRNA-mRNA regulatory network and the exploration of their regulatory relationships have provided a molecular basis for the study for the treatment of DD with Baduanjin.To conclude, our research is the first systematic profiling of mRNA, lncRNA, and circRNA in patients with DD undergoing Baduanjin intervention. We demonstrated that a 12-week Baduanjin intervention could significantly reduce blood glucose, SDS, and PHQ9 scale scores in patients with DD. These effects may be achieved by regulating the expression of the non-coding RNAs. This study hints towards an epigenetic role of Baduanjin intervention in DD and provides valuable insights in its potential mechanism.Additional file 1. Minimum Standards of Reporting Checklist.Additional file 2. Differentially expressed mRNAs.Additional file 3. Differentially expressed circRNAs.Additional file 4. Differentially expressed lncRNAs.


CYLD, promotor methylation of the Wnt antagonists WIF1, DKK1, DKK3, and sFRP1, sFRP2, sFRP4, sFRP5, as well as overexpression of the co-transcriptional activator BCL9 and the R-spondin receptor LGR4. Furthermore, Wnt activity in MM is strongly promoted by interaction of both Wnts and R-spondins with syndecan-1 (CD138) on the MM cell-surface. Functionally, aberrant canonical Wnt signaling plays a dual role in the pathogenesis of MM: (I) it mediates proliferation, migration, and drug resistance of MM cells; (II) MM cells secrete Wnt antagonists that contribute to the development of osteolytic lesions by impairing osteoblast differentiation. As discussed in this review, these insights into the causes and consequences of aberrant Wnt signaling in MM will help to guide the development of targeting strategies. Importantly, since Wnt signaling in MM cells is largely ligand dependent, it can be targeted by drugs/antibodies that act upstream in the pathway, interfering with Wnt secretion, sequestering Wnts, or blocking Wnt (co)receptors.Aberrant activation of Wnt/β-catenin signaling plays a central role in the pathogenesis of a wide variety of malignancies and is typically caused by mutations in core Wnt pathway components driving constitutive, ligand-independent signaling. In multiple myelomas (MMs), however, these pathway intrinsic mutations are rare despite the fact that most tumors display aberrant Wnt pathway activity. Recent studies indicate that this activation is caused by genetic and epigenetic lesions of Wnt regulatory components, sensitizing MM cells to autocrine Wnt ligands and paracrine Wnts emanating from the bone marrow niche. These include deletion of the tumor suppressor  CCND1,11q13), FGF3/MMSET (4p16.3), Cyclin D3 , MAFC (16q23), and MAFB (20q11). Most of the remaining cases are hyperdiploid, with recurrent trisomies of chromosomes 3, 5, 7, 9, 11, 15, 19, and 21 [KRAS, NRAS, BRAF, and P53, genetic and epigenetic abnormalities in NF kappa B and Wnt pathway components, and genetic aberrations in the MYC family of oncogenes [Multiple myeloma (MM) is a clonal expansion of malignant plasma cells in the bone marrow (BM) that is typically accompanied by paraproteinemia, lytic bone lesions, hypercalcemia, cytopenias, and renal failure. It is preceded by monoclonal gammopathy of undetermined significance (MGUS), an asymptomatic premalignant accumulation of clonal plasma cells that shares genetic features with MM . In ~50%CCND3, 6p1, MAFC -6, insulin-like growth factor (IGF), hepatocyte growth factor (HGF) and a proliferation-induced ligand (APRIL), which collectively provide signals essential for growth and survival . Both noWnt genes in the human genome which encode lipid-modified secreted glycoproteins, acting as ligands for their cognate Frizzled (FZD) receptors. Wnts are relatively unstable and insoluble due to their hydrophobic nature, which constrains long-range signaling. As a consequence, they act as typical niche or stem cell factors [PORC), and is required for signaling and secretion [2+ signaling pathway. Whereas some members of the Wnt ligand and Fzd receptor families are strictly dedicated to either canonical or non-canonical signaling, most Wnts and Fzds are promiscuously involved in both pathways. In this review, we will mainly focus on canonical Wnt signaling.The Wnt cascade represents a highly conserved developmental signal-transduction pathway involved in a variety of cellular processes, including regulation of proliferation, cell-fate, migration, and cell polarity. There are 19  factors , 13. Theecretion , 15. WntMYC and CCND1 (encoding Cyclin D1) [Canonical Wnt signaling revolves around the stabilization and subcellular localization of the transcriptional coactivator β-catenin Fig. . In the clin D1) , 20.Fig.2+ pathway, binding of a Wnt ligand to its receptor results in the release of calcium ions from the endoplasmic reticulum (ER) via the activation of G-proteins, phospholipase C (PLC), and phosphodiesterase (PDE). In turn, elevation of intracellular calcium levels activates enzymes such as protein kinase C (PKC), resulting in altered cell motility [In contrast to canonical Wnt signaling, non-canonical Wnt signaling is independent of LRP5/6 and β-catenin and plays an important role in regulating cell polarity, adhesion, and migration. In Wnt/PCP signaling, engagement of a Wnt ligand to a Fzd receptor results in activation of the small GTPase RhoA and downstream protein kinases, including Rho-associated protein kinase (ROCK), which regulates cytoskeletal dynamics by dictating the localization of structural proteins such as actin –23. In tmotility .ZNRF3/RNF43, as well as gene fusions of R-spondin causing overexpression, have recently been identified in a number of Wnt-driven tumors, including colorectal cancer (CRC) and pancreatic ductal adenocarcinoma [Deregulation of canonical Wnt signaling plays a central role in malignant transformation and tumorigenesis in a variety of tissues. This is exemplified by the intestinal epithelium. In this archetypal Wnt-dependent tissue Wnt signaling is required for stem cell maintenance and, when hyperactivated, is the main driver of malignant transformation. Oncogenic Wnt signaling typically results from mutations in core pathway components such as APC, β-catenin, and AXIN, causing constitutive, ligand-independent signaling . In addiarcinoma –32. ThesCCND1 (encoding cyclin D1) and MYC, which are known to play a key role in the pathogenesis of MM [AURKA), which was identified as an important effector of Wnt-driven proliferation in MM [Initial evidence for a role of canonical Wnt signaling in the pathogenesis of MM came from a study by Derksen et al.  demonstris of MM , 42, 43.on in MM . While tAPC, CTNNB1 (encoding β-catenin) and RNF43 that are recurrently found in other Wnt-driven tumors  maintenance  and earlCDKN2A) [APC or at least one of the secreted Wnt antagonists WIF1, DKK1, DKK3, and sFRP1, sFRP2, sFRP4, and sFRP5 was detected in the majority of pMMs and HMCLs and was associated with advanced disease stage  . InteresEXT1), which is required for heparan sulfate chain synthesis, mitigated binding of both Wnts and R-spondins to MM cells and attenuated Wnt pathway activation. Moreover, silencing of EXT1 attenuated the growth of MM tumors both in vitro and in vivo, which could be rescued by ectopic expression of a constitutive active β-catenin mutant or by c-MYC expression, indicating involvement of canonical Wnt signaling [Interestingly, we recently reported that syndecan-1 (CD138) acts as a key mediator of both R-spondin/LGR4 signaling and signaling by Wnt ligands in MM . Syndecaignaling , 67.membrane proximal regulators of the Wnt signaling cascade. However, aberrant expression of intracellular Wnt pathway regulators CYLD and BCL9 can also strongly promote Wnt signaling in MM. Loss of the tumor suppressor CYLD, by gene deletion and/or inactivating mutations, is among the most common genetic abnormalities in MM and is associated with a poor prognosis [CYLD mRNA expression and the strength of a Wnt target gene signature. In addition, gene-set-enrichment analysis showed enrichment of Wnt pathway genes in CYLD low compared to CYLD high MM patients. These data suggest that loss of CYLD facilitates Wnt signaling in MM cells. Indeed, functional studies demonstrated that silencing of CYLD in MM cells enhances β-catenin stabilization and β-catenin/TCF-mediated transcription, while reintroduction of CYLD in MM cells that display a bi-allelic loss of the CYLD locus suppresses Wnt activity and impairs proliferation. Thus, loss of CYLD can promote Wnt pathway activation and drive proliferation in MM [The above described epigenetic alterations enhance the sensitivity of MM cells to Wnt ligands by affecting rognosis , 68–72  or mutations in the Wnt coreceptor LRP5, which prevent binding of the Wnt antagonists sclerostin and DKK1, have been shown to cause increased bone mass. Conversely, loss-of-function mutations of LRP5 have been identified in osteoporosis-pseudoglioma syndrome (OPPG), which is characterized by decreased bone mass . A landmntiation , 85.DKK1 and sFRP3/FRZB expression levels were significantly correlated to the presence of lytic bone disease [In addition to DKK1, the secreted Wnt inhibitors sFRP2 , sFRP3/F disease . Intrigu disease , 62, 65. disease  or by sy disease  or an an disease , 93 was  disease . Ectopic disease  or with  disease , 93 was  disease , 92, 94  disease .Fig. 3ScAs discussed in this review, current evidence indicates that canonical Wnt signaling plays a dual and disease stage-specific role in the pathogenesis of MM. At early disease stages when MM cells are strictly confined to the BM, they secrete various Wnt antagonists, which attenuate osteoblast differentiation and thereby contribute to the development of osteolytic lesions. Since osteoblast precursors secrete high levels of cytokines and growth factors that promote MM cell growth and survival this creates a tumor permissive environment Fig. . During CCND1, MYC, and AURKA and causes drug resistance. Of note, deregulation of the cell cycle is a unifying pathogenic event required for malignant transformation of plasma cells [Oncogenic Wnt activity is typically driven by mutations in key Wnt pathway components, but these pathway intrinsic mutations are rare in MM Table . Insteadma cells .Given the role of DKK1 in the development of lytic bone disease, a mAb targeting DKK1 has been developed and has been tested in clinical trials. Initial reported results indicate that anti-DKK1 treatment is well tolerated; thus far effects on MM bone disease and disease outcome have not been reported . ImportaIn addition to canonical Wnt signaling, several studies have indicated a role of non-canonical Wnt signaling in MM pathogenesis. Activation of this signaling cascade involves signaling true RhoA/ROCK and PKC family members, and mediates migration, invasion, and adhesion of MM cells, resulting in tumor dissemination and CAM-DR.The central role of the Wnt pathway in a wide variety of malignancies has led to the development and (pre)clinical testing of many monoclonal antibodies, and small molecule inhibitors, targeting the Wnt signaling cascade at multiple distinct levels Fig.  30, 80,–109. Des


HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) is a progressive neurological and inflammatory disease, associated with HTLV-1 infection. HAM/TSP neurological disease is a consequence of an inflammatory reaction, and adaptive immune responses, through the secretion of anti-inflammatory and pro-inflammatory cytokines, play an important role in the outcome of infection and disease progression. Studies addressing the association between cytokines functional single nucleotide polymorphisms and HAM/TSP development are scarce.n = 68) and in asymptomatic HTLV-1 positive carriers (n = 83) from Rio de Janeiro, Brazil, in a case-control study. HTLV-1 infected patients were genotyped for SNPs in five cytokine genes: TNFA-308G/A, IL6-174G/C, IFNG + 874 T/A, TGFB at the codons + 10 T/C and + 25G/C, IL10-592C/A and -819C/T, and -1082A/G and proviral load (PVL) was quantified. Associations between genotypes, haplotypes, clinical outcome and pro viral load were evaluated.The genetic polymorphisms of cytokine genes were evaluated in HAM/TSP patients (TNFA-308GG, IL6-174GG/GC, IL10-592AA and -819CC and TGFb1 high producers phenotypes were correlated with higher PVL in HAM/TSP patients versus asymptomatic carriers.Lack of association between the cytokine polymorphisms and disease outcome was observed. The genotypes We did not observe association between cytokine polymorphisms and risk for HAM/TSP development in Brazilian HTLV-1 infected individuals, regardless of differences in PVL between HAM/TSP versus asymptomatic carriers in specific cytokine polymorphisms. The HTLV-1 virus is the etiological agent of two major diseases: adult T cell leukemia and the neurological disease HTLV-associated myelopathy / tropical spastic paraparesis (HAM/TSP), a progressive neurological and inflammatory disease of the central nervous system. Although many of the infected individuals are asymptomatic, approximately 2–5% of the infected individuals will develop HAM/TSP  × 100  × 100 .p-values < 0.05 statistically significant. Nonparametric data were evaluated by Kruskal-Wallis and Mann-Whitney tests and a Spearman correlation, and association between qualitative variables  was tested by chi-square, Cochran-Armitage or Fisher’s exact test. P-values from tables with any significant result were corrected using Bonferroni correction for multiple comparisons. Results with a p-value < 0.05 were considered statistically significant. All analyses were performed using the software GraphPad Prism 6.0 or R version 3.1.0  p = 0.01), predisposing to disease outcome, although the data lost significance after Bonferroni correction for multiple comparisons. Low-producer haplotypes  have been previously associated with AC or HAM/TSP compared with healthy controls [IL10–819CC and -592AA carriers, presenting high levels of IL10 production, have the highest PVL when the HAM/TSP group was compared with the AC group.HTLV-1 infection in HAM/TSP patients does not alter the expression of IL-10 when compared with AC patients . SNPs in HAM/TSP ; however HAM/TSP . No assopulation  in a higpulation . In our pulation  there waic model . There wcontrols ; howeverIL10 -592C/A and IL10–1082A/G, and HTLV-1 infection in some specific populations. On the other hand, some polymorphisms, such as TNF -308G/A, IFNG + 874 T/A, IL6 -174G/C, are repeatedly not associated with the disease, regardless genetic background population, suggesting irrelevant role in the HTLV-1 infection. However, more studies in different populations are needed to confirm this hypothesis. Despite no association with disease development, some of these polymorphisms in cytokine genes, such as IFNG + 874 T/A, TNF -308G/A, IL6 -174G/C, TGFB + 10 T/C and + 25G/C, IL10–819C/T and -592C/A might change PVL, indicating a role of this polymorphisms in the controlling of the viral replication and disease prognosis.The infection caused by HTLV-1 elicits a robust immune response with many factors affecting the cytokines gene expression, such as genetic population’s background, virus subtypes, immunomodulation and individual health status. Indeed, it has been shown that different subtypes of HTLV-1 are region restricted, besides the genetic background of ethnicities across the world . This faOur study might be analyzed considering its limitations such as the number of patients enrolled and the unavailability to measure cytokines concentrations in plasma or cultured cells in the patients. As we used convenience samples, we had limitations regard sample size and biological material to perform the analysis. Despite these polymorphisms are well established to modulate the cytokines production, is not clear its role on the HTLV-1 infection. Future studies addressing the role of these polymorphisms in different populations, associated with the cytokines production, might clarify the role of these functional SNPs in the HTLV-1 infection.TNFA-308G/A, IL6-174G/C, IFNG + 874 T/A, TGFB at the codons + 10 T/C and + 25G/C, IL10-592C/A and -819C/T, and -1082A/G polymorphisms are related to either disease progression, even those related with the amount of secreted cytokines. Genetic background may be studied in other sets of populations to determine and understand the complex role of cytokine networks in HTLV-1 infection and improve the clinical studies for this disease.In conclusion, besides the importance given to pro- and anti-inflammatory cytokines in the outcome of HAM/TSP, we could not affirm that