Chronic hepatitis C virus (HCV) infection is a major cause of liver cirrhosis and hepatocellular carcinoma (HCC) which are leading indications of liver transplantation (LT). To date, there is no vaccine to prevent HCV infection and LT is invariably followed by infection of the liver graft. Within the past years, direct-acting antivirals (DAAs) have had a major impact on the management of chronic hepatitis C, which has become a curable disease in the majority of DAA-treated patients. In contrast to DAAs that target viral proteins, host-targeting agents (HTAs) interfere with cellular factors involved in the viral life cycle. By acting through a complementary mechanism of action and by exhibiting a generally higher barrier to resistance, HTAs offer a prospective option to prevent and treat viral resistance. Indeed, given their complementary mechanism of action, HTAs and DAAs can act in a synergistic manner to reduce viral loads. This review summarizes the different classes of HTAs against HCV infection that are in preclinical or clinical development and highlights their potential to prevent HCV infection, e.g., following LT, and to tailor combination treatments to cure chronic HCV infection. Flaviviridae family. It is a highly variable virus that has been classified into six major genotypes [Hepatitis C virus (HCV) is a hepatotropic enveloped RNA virus of the enotypes . Approxienotypes ). Severai.e., DAAs, or interfere with cellular factors involved in viral infection, i.e., host-targeting agents (HTAs). For many years, HCV-host interactions have remained largely elusive due to the lack of robust model systems, and anti-HCV drug development has paralleled discoveries about the molecular mechanisms of viral replication. Indeed, the establishment of increasingly sophisticated model systems to study HCV infection  hasiewed in ,7,8,9) aiewed in ). While iewed in ,12,13. TThe HCV life cycle can be subdivided into five different steps, including viral entry, viral translation, viral replication, viral assembly, and release of new virions. Each of these steps requires defined virus-host interactions to ultimately enable the virus to establish chronic infection. Given the amount of host factors involved in the HCV life cycle, this review solely focuses on some of these host factors that have been suggested to represent potential targets for HTAs during distinct steps of the HCV life cycle within hepatocytes . Other cLentinula edodes mycelia solid culture extract, that has been reported to exhibit hepatoprotective activity, might inhibit HCV attachment through binding to apoE on the viral particle [The HCV entry process has been particularly well characterized within the past years (for a review see ). The inparticle  given thparticle . Interesparticle ,68. In aparticle ,69. Mostparticle .via clathrin-mediated endocytosis. It is believed that the interaction of the HCV envelope glycoproteins E1 and E2 with SR-BI, CD81, CLDN1 and potentially OCLN plays a major role in this process and some of these host factors have thus been suggested to play the role of “HCV receptors” [via induction of IFN-α inducible protein 6 (IFI6), an IFN-stimulated gene (ISG) [Following initial attachment to the hepatocyte basolateral membrane, the virus interacts with additional host factors expressed at the hepatocyte cell surface to ultimately enter its host cell ceptors” ,73,74,75ceptors” . Of noteceptors” ,77,78,79ceptors” ). Thus, ceptors” ,94,95,96ne (ISG) .via clathrin-mediated endocytosis, the viral genome is released into the cytoplasm after fusion of viral and endosomal membranes. In addition to some of the compounds described above, HCV endocytosis and/or fusion can be efficiently perturbed using amphipathic DNA polymers and small molecules including two anti-malaria drugs , arbidol and sylimarin/silibinin [Following internalization ilibinin ,102,103 ilibinin ,104,105.in vitro infectivity remain sensitive to SR-BI-specific antibodies in vivo, as demonstrated using the human liver-chimeric uPA-SCID mouse model [In addition to cell-free virus entry, HCV has been shown to also directly infect neighboring hepatocytes via a process called cell-to-cell transmission ,107,108.se model .https://clinicaltrials.gov/; Identifier: NCT01835938). Furthermore, silymarin/silibinin-derived compounds have been evaluated in phase 2/3 clinical trials and (case) reports suggest their efficacy in decreasing viral loads in chronic HCV patients and prevention of hepatitis C recurrence after LT, although other studies reported viral rebounds [https://clinicaltrials.gov/; Identifier: NCT02058173), no clinical trial of ferroquine in hepatitis C patients has been registered so far.Host-targeting entry inhibitors (HTEIs) including antibodies directed against host entry factors or co-factors as well as small molecule inhibitors have proven to efficiently prevent/delay HCV infection in the human liver-chimeric uPA-SCID mouse model ,110,111 rebounds ,114,115.via a cap-independent mechanism. The HCV IRES binding to the 40S ribosomal subunit is independent of canonical or non-canonical initiation factors. Moreover, HCV translation initiation only requires eukaryotic initiation factors (eIF) eIF2, eIF3 and eIF5 [Following viral endocytosis and fusion, HCV RNA is translated in an internal ribosome entry site (IRES)-mediated manner. The highly conserved IRES is located in the 5′-untranslated region (5′-UTR) of the viral genome ,117. HCVand eIF5 ,119. In and eIF5 .in vivo efficacy of molecules interfering with HCV IRES-ribosome association remains to be determined.IRES-ribosome interaction has been shown to be mediated through the binding of the 40S subunit to the stem loop III, domain e and f (SLIIIef), of the HCV IRES. A recent study demonstrated that a small RNA mimicking the SLIIIef domain (SLRef) selectively inhibits HCV translation by retaining the ability to interact with the 40S subunit. Interestingly, in line with previous studies, it was shown that the anti-HCV activity of SLRef could be mainly due to interaction and sequestration of the La antigen . RibosomFurthermore, microRNA-122 (miR-122) plays an important role in HCV translation. MiR-122, one of the most abundant liver-expressed miRNAs, binds to the HCV genome and enhances viral translation and replication ,124. SeqRecently, the receptor for activated C kinase 1 (RACK1), a component of the ribosomal 40S subunit ,126, wasFollowing translation and cleavage of the HCV polyprotein, the viral replicase complex, composed of HCV non-structural (NS) proteins, NS3, NS4A, NS4B, NS5A and NS5B, is assembled. HCV replication is a highly coordinated process involving host factors in addition to the replicase complex. Several functional small interfering RNA (siRNA) and chemical screens have been performed to uncover host factors involved in HCV replication and have provided useful insights into this process ,134,135.Ppia gene had been knocked out [HCV has been demonstrated to remodel intracellular membranes, including endoplasmic reticulum (ER) membranes, for its replication and this process takes place at the so-called “membranous web” . Indeed,cked out . In addicked out ,149,150.cked out ). Cyp incked out ,151. Thecked out ,153,154.cked out .in vitro have been developed, but their in vivo toxicity as well as the lethal phenotype of transgenic mice exhibiting kinase-defective PI4KIIIα preclude their further clinical development as antivirals [in vitro by treatment with inhibitors of 3-hydroxy-3-methylglutaryl CoA (HMGCoA) reductase or with an inhibitor of protein geranylgeranyl transferase I [in vitro studies have provided novel insights into the mechanism of action of fluvastatin. The anti-HCV effect of fluvastatin has been suggested to be due to the induction of microtubule bundling and a decrease of expression of the microtubule binding partner doublecortin-like kinase 1 (DCLK1) [NS5A is a central player within the membranous web as it interacts with a plethora of host factors, thereby orchestrating viral replication. The lipid kinase phosphatidyl-inositol-4-kinase III alpha (PI4KIIIα) is one of the major NS5A partners in this process . Indeed,tivirals ,160,161.tivirals . Particutivirals . HCV repferase I ,165. Notferase I . Fluvastferase I ,167,168. (DCLK1) . Further (DCLK1) ,172,173. (DCLK1) ,175. In  (DCLK1) . Interes (DCLK1) , further (DCLK1) . Indeed, (DCLK1) . ESRα ma (DCLK1) ,178,179. (DCLK1) ,180. The (DCLK1) ,179.In addition to its role in HCV genome translation as described above, miR-122 is a hepatocyte-specific host factor essential for HCV replication. Indeed, HCV cannot replicate in cells lacking miR-122 or in which miR-122 has been sequestered ,184,185.Besides inhibiting host factors required for viral replication, HCV replication may also be inhibited by modulating host factors that restrict HCV replication, e.g., the elongation factor Tu GTP binding domain containing 2 (EFTUD2) and the human cytidine deaminase APOBEC3G (hA3G) ,191,192.Interestingly, compounds inhibiting histone deacetylase 6 (HDAC6) have also been reported to inhibit HCV replication ,195. Thein vitro [https://clinicaltrials.gov/; Identifier: NCT01387958), but additional studies are needed to assess its antiviral efficacy in patients.A remarkable hallmark of HCV is its association with VLDL/low density lipoprotein (LDL) to form an infectious lipoviroparticle (LVP) . Intracein vitro . The safCore recruitment to LDs also depends on the composition and the fluidity of ER lipid bilayers that is regulated by the cytosolic phospholipase A2, group 4A, (PLA2G4A) and upstream mitogen-activated protein kinases-extracellular signal regulated kinases (MAPK/ERK). PLA2G4A has been shown to contribute to HCV assembly since pyrrolidine-2 (Py-2), a chemical inhibitor of PLA2G4A, induces aberrant HCV particle production . FurtherDirectly targeting core trafficking in hepatocytes also represents an interesting antiviral strategy given the numerous host factors involved in this process . These iOther HTAs interfering with host cell lipid metabolism have been reported to display antiviral activity against HCV. Indeed, resveratrol that increases the expression of PPARα, a nuclear transcription factor that activates fatty acid oxidation and is decreased by the core protein ,215, impBesides core, NS5A is another major player within the HCV replication cycle that connects replication and assembly. NS5A-core interaction on LDs is a crucial step for viral assembly ,219,220.via fusion with resident apoE containing LDs . Accumuiew, see ,230,231,iew, see ,225,232.iew, see  . HoweveThe VLDL secretory pathway is also hijacked to facilitate HCV egress from infected hepatocytes ,225 Fig. VLDL sein vivo, making these molecules promising candidates for clinical trials [To counteract HCV proliferation, several studies reported a novel antiviral strategy that uses the human heat shock cognate protein 70 (Hsc70) as an antiviral target ,239,240.l trials ,239. Recl trials . Howeverin vitro. Moreover, celgosivir administration in combination with PEG-IFN α and ribavirin displayed a clinical benefit in genotype 1 chronically infected patients . Long-ti.e., a Cyp inhibitor [in vitro [in vitro, viral rebound is usually observed following withdrawal of the drug. However, the addition of an HTEI to the cell culture at the time of DAA withdrawal allows the further decrease of the viral load, indicating that HTEIs limit viral rebound from DAA therapy [in vitro data, the Cyp inhibitor alisporivir, currently in phase 2/3 clinical trials, has been shown to efficiently reduce viral loads in chronic hepatitis C patients in combination with ribavirin [Given that they target a highly variable virus, the main limitation of DAAs is viral resistance. Viral resistance may be prevented using different strategies including (i) targeting conserved viral sequences less prone to mutation; (ii) using a combination of drugs exhibiting distinct targets and mechanisms of action; and (iii) increasing host antiviral responses. In contrast to the first DAAs, the emergence of viral resistance with recently approved DAAs, such as sofosbuvir, is less frequent but has nevertheless been described ,244,245.nhibitor . Furtherin vitro ,246,247. therapy ,246. Furibavirin  and futuin vitro [in vitro [in vitro. While most mutants were still dependent on Cyp and/or miR-122, specific mutations in NS5A eliminated the dependence of HCV RNA replication on the expression of host CypA [in vivo where CLDN1 is expressed at much higher levels than CLDN6 and CLDN9 remains to be determined .During the last decade, the development of new DAAs considerably improved anti-HCV therapy, displaying high cure rates in treated patients. However, treatment failure and resistance in a subset of patients remain a challenge. Alternative approaches targeting host factors involved in HCV pathogenesis offer new treatment opportunities. Due to low genetic variability of host factors, HTAs exhibit a high genetic barrier to resistance and display a pan-genotypic antiviral activity.Several HTAs are currently in development in preclinical or clinical trials . Proof-o


Magnaporthe oryzae is the most devastating disease of rice and poses a serious threat to world food security. In this study, the distribution and effectiveness of 18 R genes in 277 accessions were investigated based on pathogenicity assays and molecular markers. The results showed that most of the accessions exhibited some degree of resistance . Accordingly, most of the accessions were observed to harbor two or more R genes, and the number of R genes harbored in accessions was significantly positively correlated with RF. Some R genes were demonstrated to be specifically distributed in the genomes of rice sub-species, such as Pigm, Pi9, Pi5 and Pi1, which were only detected in indica-type accessions, and Pik and Piz, which were just harbored in japonica-type accessions. By analyzing the relationship between R genes and RF using a multiple stepwise regression model, the R genes Pid3, Pi5, Pi9, Pi54, Pigm and Pit were found to show the main effects against M. oryzae in indica-type accessions, while Pita, Pb1, Pik, Pizt and Pia were indicated to exhibit the main effects against M. oryzae in japonica-type accessions. Principal component analysis (PCA) and cluster analysis revealed that combination patterns of major R genes were the main factors determining the resistance of rice varieties to M. oryzae, such as ‘Pi9+Pi54’, ‘Pid3+Pigm’, ‘Pi5+Pid3+Pigm’, ‘Pi5+Pi54+Pid3+Pigm’, ‘Pi5+Pid3’ and ‘Pi5+Pit+Pid3’ in indica-type accessions and ‘Pik+Pib’, ‘Pik+Pita’, ‘Pik+Pb1’, ‘Pizt+Pia’ and ‘Pizt+Pita’ in japonica-type accessions, which were able to confer effective resistance against M. oryzae. The above results provide good theoretical support for the rational utilization of combinations of major R genes in developing rice cultivars with broad-spectrum resistance.Rice blast caused by  Oryza sativa L.) is one of the major food crops worldwide, feeding more than half of the world’s population [Magnaporthe oryzae is the most severe disease of rice worldwide. Due to its widespread distribution and ability to survive in a wide range of environmental conditions, yield losses between 10% and 30%, i.e. destroying rice that could have fed more than 60 million people [R genes into elite rice varieties is the most economical and environmentally friendly way to protect crops against this disease [Rice  [pi21 and Pb1) [R genes (except Pid2 and pi21) belong to the nucleotide-binding site and leucine-rich repeat (NBS–LRR) class of R genes [R genes such as Pi5, Pia/Pi-Co39, Pikm, Pik and Pikp require two adjacent NBS-LRR class genes for full functionality [Pid2 encodes a B-lectin receptor kinase [pi21 encodes a proline-rich protein that containing a putative heavy metal-binding domain and putative protein-protein interaction motifs [R genes has advanced our understanding of the molecular basis of blast resistance in rice, we know little about how these cloned R genes are distributed in modern cultivated rice varieties. Additionally, information both on the resistance effects of R genes in different genetic backgrounds and on which genes are more effective in breeding practice is also missing. Furthermore, there are more than 100 R genes in the rice genome, which means that a single germplasm may harbor several R genes in various combinations, but the fact that resistance reactions show significant differences between germplasms indicates that these variations could be caused by different R gene combinations. Therefore, the question arises of which R genes could be combined to provide a favorable resistance effect across multiple backgrounds. Addressing this question will be helpful in the improvement of blast resistance breeding programs.To date, more than 100 rice blast resistance genes have been identified in rice ,5, and 2and Pb1) ,12 have  R genes , and somionality ,14. Hower kinase , while pn motifs . AlthougR gene in diverse elite germplasm is the first step for utilization of R genes in rice breeding programs. The conventional methodology to identify allelic variation based on phenotype is limited by large workload and time-consuming, as well as the strong dependence on environmental conditions. However, for R genes that have been isolated, it is now possible to replace phenotypes with molecular markers as the basis for defining alleles [R genes, such as Pi1 [Pi2 [Pi5 [Pi9 [R genes in gene introgression and gene pyramiding. Moreover, the identification of functional markers (FMs), which include single nucleotide polymorphisms (SNPs) and insertion/deletions (InDels), were derived from polymorphic sites within genes causally involved in phenotypic trait variation and is particularly useful in several genetic backgrounds [R genes, such as Pita [Pib [Pit [Pid3 [Accurate identification of a particular  alleles . Over thh as Pi1 , Pi2 [17Pi2 [Pi5 , Pi9 [19kgrounds . Some FM as Pita , Pib [22Pib [Pit  and Pid3it [Pid3  have beeR genes was analyzed in 277 accessions of Chinese elite rice parental lines and the donors of R genes using FMs and LMs, and the resistance frequency (RF) in the 277 accessions was evaluated by a total of 76 isolates. The R genes showing the main effects to M. oryzae in indica and japonica rice genome were identified through multiple stepwise regression analysis. The combination patterns of major R genes in the indica-type and japonica-type rice germplasm were ascertained based on principal component analysis (PCA) and cluster analysis of various gene combination factors. The results of this study are expected to guide the deployment and combination of R genes and provide useful information to improve blast resistance.In the present study, the distribution of 18 japonica and 149 indica) mainly consisting of Chinese elite rice parental lines, were analyzed in the present study , Toride 1 (Pizt), K59 (Pit), Kanto 51 (Pik), Tsuyuake (Pikm), K60 (Pikp), BL 1 (Pib), Yashiromochi (Pita), K3 (Pi54), Shin 2 (Piks) and Pi No.4 (Pita2). The second set consisted of Chinese differentials, including Tetep , Zhenglong 13 (unknown), Sifeng 43 , Dongnong 363 , Kanto 51 (Pik), Hejiang 18 (Pia) and Lijangxituanheigu (LTH); the third set consisted of IRRI differentials, including six resistant near isogenic lines (NILs) in the genetic background of Co39 [Pi9, Pi40, Pigm, Pid2 and Pid3 genes, the donor lines 75-1-127 [A total of 277 accessions  with 0.02% Tween-20 as described by Huang et al. [To evaluate blast resistance, a total of 76 isolates collected from Sichuan (SC), Anhui (AH), Jiangsu (JS), Hainan (HN), Hubei (HB), and Zhejiang (ZJ) provinces in China from 2010–2012 were examined in this study . The plag et al. . The inoM. oryzae could be calculated according to the following formula: RF =  *100%. Each RF value was assessed from two technical replicates.Disease reactions were assessed 7 days after inoculation, and lesions on the rice leaves were evaluated on a scale of 0 to 5 according to standard procedures , where 0R genes, including six alleles of the Piz locus  [Pik and Pi1) [R genes  [A total of 15 FMs and 9 LMs were selected from published primer sequences to detect 18 nd Pigm) , 36, twoand Pi1) , 37 and PCR amplification was conducted following a standard protocol , includiR genes and RF was analyzed using a multiple stepwise regression model, which is an available option in Matlab (V.7.0) software, according to Xu et al. [R genes that were significantly correlated with RF (P≤0.01) were subjected to various R gene combination factors according to factor analysis  in SPSS (V.21) software. Based on various R gene combination factors, the germplasms were clustered into several groups. Each group represents one class of R gene combination patterns found among the germplasms. Significant differences in the RF between population groups were analyzed by one-way analysis of variance (ANOVA) [The relationship between u et al. . R genes (ANOVA) R genes, including Pia, Pib, Pit, Pish, Pi5, Pi54, Pita, Pid2, Pid3, and Pb1, were easily identified based on their FMs or LMs  from each other, several markers were selected from published primer sequences. An InDel polymorphism specific for Pi9 donor 75-1-127 was obtained using the marker F9, which discriminates Pi9 from other alleles , which means that the greater the number of R genes found in the accessions, the higher the RF against M. oryzae.Most of the accessions harbored more than one of the 18 detected cessions . The num–5 genes . CompareR genes were detected in 277 accessions using a set of 24 FMs and LMs, and the results demonstrated that most of these R genes were present in both the indica and japonica sub-species genomes. With the exception of the R genes Pi2, Pi40, Pita, Pib and Pi54 which were evenly distributed in the indica-japonica accessions, the other R genes were found to show a disequilibrium distribution between the indica and japonica sub-species genomes. For example, the R genes Pigm, Pi9, Pi5 and Pi1 were only found in indica-type accessions, and Pik and Piz were just present in japonica-type accessions. The distribution frequency of the R genes Pit, Pid2, Pid3 and Pia was higher in indica-type accessions than in japonica-type accessions, but the distribution frequency of the R genes Pizt, Pb1 and Pish was higher in japonica-type accessions than in indica-type accessions  of the observed phenotypic variance, with partial correlation coefficients of 0.273, 0.271, 0.194, 0.190, 0.171 and 0.166, respectively  of the observed phenotypic variance overall, with partial correlation coefficients of 0.468, 0.384, 0.268, 0.243 and 0.197, respectively , ‘Pid3, Pigm’ (R2 = 20.54%) ‘Pi9’ (R2 = 16.82%) and, according to the principal component analysis results , and the 149 indica accessions were further categorized into 6 groups based on cluster analysis with R gene combination factors  and showed RF values of 80.03%, 89.83% and 77.15% respectively. Cluster V was mainly composed of R gene combination factors (Pid3 and Pigm) and exhibited RF values of 95.16%. Cluster VI included the R gene combination factor Pi9 and showed an RF of 90.51%. The RF values of clusters II, III, IV, Vand VI were significantly higher than that of cluster I, suggesting that major R genes could significantly improve the resistance of the rice germplasm to M. oryzae (P<0.01). Furthermore, the results showed that there were significant differences in the level of resistance between clusters that contained various R gene combination factors. For example, cluster II mainly contained R gene combination factors  and could be combined into 5 gene combination patterns . However, the RF of cluster II was significantly lower than that of cluster VI, which contained the R gene factor ‘Pi9’ with the combination patterns ‘Pi9+Pi54’ and ‘Pi9’, indicating that the R gene combination pattern may be the main factor determining rice variety resistance to M. oryzae in indica-background accessions.The genes that showed the main effects against  factors . Clusterf 64.84% . ClusterR gene combination factors, ‘Pizt, Pia, Pita’(R2 = 28.87%), ‘Pik’ (R2 = 21.61%) and ‘Pizt, Pia, Pb1’ (R2 = 21.11%), were categorized in japonica-type accessions among the R genes that showed the main effects against M. oryzae , and the 128 japonica-type accessions were further classified into six major groups, clusters I, II, III, IV, V and VI, based on the observed R gene combination factors  and showed RF values of 81.37% and 84.02%, respectively. Cluster V mainly included the R gene combination factor Pik, and its RF was 86.88%. Clusters II and VI mainly included R gene combination factors  and displayed RF values of 73.23% and 87.57%, respectively. The RF values of clusters III, IV, V and VI were significantly increased compared with that of cluster I (P<0.01). The resistance of cluster IV, carrying the ‘Pizt+Pia’ gene combination, and cluster VI, pyramiding ‘Pizt+Pita,’ was better than that of cluster II, carrying ‘Pia+Pb1,’ implying that various major R gene combination patterns resulted in different RFs in japonica-background accessions.Similarly, three  factors . Signifir groups . ClusterR genes have been identified and mapped in a variety of rice genotypes in previous works. Unfortunately, not all R genes are effective against pathogenic isolates of M. oryzae. Therefore, how to employ a few of these R genes to achieve broad-spectrum and durable resistance must be considered during rice blast resistance breeding.A large number of R genes in 277 germplasms, we found that the distribution of R genes was significantly different between the indica and japonica sub-species genomes. For example, the R genes Pit, Pid3, Pid2 and Pia were mainly detected in indica-type accessions, and Pigm, Pi9, Pi5 and Pi1 were only present in indica-type accessions. However, the R genes Pizt, Pb1 and Pish were mainly identified in japonica-type accessions, and Pik and Piz were restricted to japonica-type accessions. These results were possible because these R genes have experienced significant differentiation due to reproductive isolation and decreased interflow between indica-japonica sub-species during the evolutionary process. Thus, it may be an effective strategy to transfer these R genes within subspecies using marker-assisted selection (MAS) to create new resistant germplasms, for example, transferring Pid3, Pigm, Pi1 and Pi5, which originated in the indica genome, to the japonica rice genome.According to the detection results of 18 R genes also results in various R genes showing the main effects against M. oryzae between indica and japonica germplasms, such as the R genes Pid3, Pi5, Pi9, Pi54, Pigm and Pit in indica-type accessions and Pita, Pb1, Pik, Pizt and Pia in japonica-type accessions . It has been shown that pyramiding lines with ‘Pi1+Pi2’ genes shows complete resistance to the most virulent races of the pathogen originating from southern China [R gene, Pigm and Pi1, possessed a complementary effect with respect to the blast isolates and the RF of pyramiding lines with ‘Pigm+Pi1’ to blast isolates originated from China was more than 90%. In addition, it has been documented that at least nine R genes Pi1, Pi2, Pi5, Piz, Pi9, Pi40, Pizt, Pi33 and Pigm appear to confer broad-spectrum resistance to a number of isolates or races from one or more countries [Piz locus . All of these results indicate that the alleles of the Piz locus play a key role in conferring resistance to M. oryzae. Therefore, it may be an effective strategy to use different alleles of Piz locus as the backbone for pyramiding with other R genes with strong complementary effect and broad-spectrum resistance, such as Pi1, Pi5, Pi33, Pi54 and Pid3, could create new resistant germplasms and further enhance of the level and broad-spectrum of blast resistance in rice.Most of above-mentioned effective rn China . Furtherrn China  demonstrountries ,34,47, aS1 Fig(TIF)Click here for additional data file.S1 Table(XLSX)Click here for additional data file.S2 Table(XLSX)Click here for additional data file.S3 Table(XLSX)Click here for additional data file.S4 Table(XLSX)Click here for additional data file.S5 Table(XLSX)Click here for additional data file.S6 Table(XLSX)Click here for additional data file.S7 Table(XLSX)Click here for additional data file.


AbstractMonoliropus belonging to the family Caprellidae was collected from the Yellow Sea, Korea. The new species differs from Monoliropusagilis Mayer, 1903, Monoliropuskazemii Momtazi & Sari, 2013, and Triprotellaamica Arimoto, 1970 as follows: 1) gnathopod 1 subrectangular; 2) pereonites 2–3 with acute triangular processes anterolaterally; 3) mandibular palp, apical article with four simple setae subdistally. The new species is fully illustrated and extensively compared with related species. This is the first record of the genus Monoliropus from Korean waters. A key to Monoliropus species is also given.A new species of the genus  Monoliropus Mayer, 1903 is one of 57 genera belonging to the family Caprellidae. Monoliropus is closely related to Metaprotella Mayer, 1890 and Triprotella Arimoto, 1970 and commonly characterized by having biarticulate flagellum of antenna 2; triarticulate mandibular palp; pereonites 3–4 with gills; uniarticulate pereopods 3–4; well developed, 6-articulate pereopod 5; in male, abdomen with a PageBreakpair of biarticulate appendages and a pair of lobes , Incheon, Korea and the Department of Life Sciences, Dankook University (DKU), Cheonan, Korea.Specimens were collected by light trap from the subtidal zone of Bukahng Port, Mokpo-si, Korea in 2012 , NIBRIV0000309620.Body Fig.  11.7 mm Monoliropus belongs to the family Caprellidae, which has close affinities with Metaprotella Mayer, 1890 and Triprotella Arimoto, 1970 as it possesses the following characters: 1) antenna 2, peduncles without swimming seta, flagellum biarticulate; 2) mandibular palp bi- or triarticulate; 3) pereopods 3–4 reduced, uniarticulate; 4) in male, abdomen with uni- or biarticulate appendages. However, Metaprotella is distinguished from Monoliropus by setal formula 1–x–y–1 of the distal article of mandibular palp and fused pereonites 6–7. Triprotella is very similar to the genus Monoliropus; however, is discernible from Monoliropus by setal formula 1-1-1 of the distal article of the mandibular palp, uniarticulate pereopods 3–4, morphology of the gnathopod 2, and form of abdomen ; 2) maxilla 1, outer plate with six stout setal teeth ; 3) mandibular palp, distal article with four simple setae (vs. three simple setae in Monoliropusagilis and Triprotellaamica); 4) gnathopod 1, propodus subrectangular ; 5) gnathopod 1, dactylus with serrations on inner margin (vs. with serrations both margins in Triprotellaamica); 6) pereonites 2–3 with acute triangular PageBreakprocesses anterolaterally ; 7) pereopods 3–4 short ; 8) abdominal appendage uniarticulate (vs. biarticulate in Monoliropuskazemii).The genus  abdomen . The speleeae is in honor of Dr. Kyung-Sook Lee, who has contributed to knowledge of Korean caprellid Amphipoda.The specific name Bukhang Port, Jukgyo-dong, Mokpo-si, Jeollanam-do, Korea.PageBreakPageBreak


Chrysanthemum morifolium is an important floral crop that is cultivated worldwide. However, due to a lack of genomic resources, very little information is available concerning the molecular mechanisms of flower development in chrysanthemum.The transcriptomes of chrysanthemum vegetative buds, floral buds and buds were sequenced using Illumina paired-end sequencing technology. A total of 15.4 Gb of reads were assembled into 91,367 unigenes with an average length of 739 bp. A total of 43,137 unigenes showed similarity to known proteins in the Swissprot or NCBI non-redundant protein databases. Additionally, 25,424, 24,321 and 13,704 unigenes were assigned to 56 gene ontology (GO) categories, 25 EuKaryotic Orthologous Groups (KOG) categories, and 285 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, respectively. A total of 1,876 differentially expressed genes (DEGs)  were identified between vegetative buds and floral buds, and 3,300 DEGs  were identified between floral buds and buds. Many genes encoding important transcription factors  as well as proteins involved in carbohydrate metabolism, protein kinase activity, plant hormone signal transduction, and the defense responses, among others, were considerably up-regulated in floral buds. Genes involved in the photoperiod pathway and flower organ determination were also identified. These genes represent important candidate genes for molecular cloning and functional analysis to study flowering regulation in chrysanthemum.Chrysanthemum morifolium. A wide range of genes was implicated in regulating the phase transition from vegetative to reproductive growth. These results should aid researchers in the study of flower-time regulation, breeding and molecular biology in chrysanthemum.This comparative transcriptome analysis revealed significant differences in gene expression and signaling pathway components between the vegetative buds, floral buds and buds of  Gebera hybrida and Helianthus annuus in Asteraceae. Chrysanthemum morifolium, another species of Asterceae, is an important crop that is widely cultivated, and indeed, complete year-round production of chrysanthemums is necessary to meet the demands of a growing market [Asteraceae belongs to the euasterids II clade of the core eudicots, one of the largest families of flowering plants. The head-like inflorescence (capitulum) of Asteraceae resembles a single large flower, and this trait is considered to be the key innovation behind the evolutionary success of the Asteraceae . Thus, tg market . Howeverg market .CRYPTOCHROME2/FHA (CRY2), GIGANTEA (GI), TIMING OF CAB 1 (TOC1) and CONSTANS (CO) [SUPPRESSOR OF CONSTANS1 (SOC1), SHORT VEGETATIVE PHASE (SVP), LEAFY (LFY), APETALA1 (AP1), GIGANTEA (GI), AGAMOUS (AG) and PISTILLATA (PI). After the initial expression of LFY, AP1 is expressed throughout the floral meristem, and together with AP2, specifies sepal identity [AP1, AP2), which determine sepal identity, A-class genes together with B-class genes , which determine petals, B-class genes together with a C-class gene (AG), which specify stamens, and the C-class gene, which specifies carpel identity [In temperate regions, the duration of the daily light period, also called the photoperiod, is one of the most important factors controlling flowering time . In ArabANS (CO) . Some imANS (CO) , 5. Manyidentity , 6. Variidentity . The claidentity .APETALA1, SEPALLATA3, and FRUITFULL [Chrysanthemum is a typical short-day plant. Under short-day treatment, chrysanthemum undergoes a major physiological change and morphological programs as it transits from a vegetative growth period to a reproductive period and finally blooms. This process is regulated by a combination of endogenous and environmental signals. By morphological observation, three developmental periods can be identified during the developmental process from vegetative growth to reproductive growth, including vegetative bud growth, flower bud differentiation, and bud development. In chrysanthemum, only a handful of important transcription factors involved in the flower developmental pathway have been isolated and analyzed, including the homologous genes RUITFULL . TherefoTranscription expression profiling, data assembly, and analysis can provide broad and deep insights into gene regulatory networks and biological pathways because they can reveal the genes downstream of key transcription factors in related pathways . The useThe cDNA libraries of vegetative buds, floral buds and buds were sequenced using the Illumina HiSeq 2000 system. After stringent quality checks and data cleaning, 58,530,798, 41,424,050 and 54,035,520 clean reads were left for vegetative buds, floral buds and buds, respectively. The average proportion of clean reads for these libraries was approximately 93%.A total of 123,354 contigs were assembled based on the high-quality reads, with a total size of 67,027,473 bp, an N50 of 641 bp and an average contig length of 543 bp. Then, scaffolds were constructed between the contigs via the paired-end relationships between the reads. A total of 95,280 scaffolds were obtained, with an N50 of 818 bp and an average length of 721 bp. We filled the intra-scaffold gaps and constructed a non-redundant unigene set from all three of the assembled datasets using CAP3 software. Finally, a total of 91,367 high-quality unigene sequences with lengths greater than 400 bp were obtained, with an average unigene length of 739 bp .Of the 91,367 unigenes, 43,137  were aligned to the Nr protein (date 2014.03) and Swissprot protein databases (date 2014.03) using an E-value threshold of < 1e-5, which meant that 48,230 unigenes  had no Swiss-Prot annotations because of missing chrysanthemum genome and expressed sequence tag (EST) information .The international standardized gene functional classification system Gene Ontology (GO) provided three ontologies—molecular functions, cellular components and biological processes—that were useful for gene annotation and analysis. Based on the Nr annotation and Swissprot protein databases, 25,424 unigenes were classified into 56 functional GO categories using Blast2GO software . A totalGIGANTEA etc. , ‘regulation of flower development’ (GO:0009909), ‘photoperiodism, flowering’ (GO:0048573), ‘positive regulation of flower development’ (GO:0009911), ‘regulation of photoperiodism, flowering’ (GO:2000028), ‘negative regulation of flower development’ (GO:0009910), ‘regulation of long-day photoperiodism, flowering’ (GO:0048586), ‘long-day photoperiodism, flowering’ (GO:0048574), ‘negative regulation of long-day photoperiodism, flowering’ (GO:0048579), ‘flower calyx development’ (GO:0048464), ‘short-day photoperiodism, flowering’ (GO:0048575), ‘flower morphogenesis’ (GO:0048439) and ‘regulation of short-day photoperiodism, flowering’ (GO:0048587). In total, 113 unigenes were assigned to these 13 terms, of which 86 were annotated as uncharacterized or predicted proteins and the remainder showing homology to F-box family genes and TEA etc. .As a eukaryote-specific version of the Clusters of Orthologous Groups (COG) tool, euKaryotic Orthologous Groups (KOG) is used to identify orthologous and paralogous proteins, providing a way to identify Joint Genome Institute (JGI)-predicted genes by KOG classification or ID. The annotated sequences were further searched for genes involved in KOG classifications for evaluating the completeness of our transcriptome library and the effectiveness of our annotation process. Of 43,137 Nr hits, 24,321 sequences were assigned to KOG classifications. Among the 25 KOG categories, the cluster for ‘Signal transduction mechanisms’  represented the largest group, followed by ‘General function prediction only’  and ‘Posttranslational modification, protein turnover, chaperones’ . The ‘Nuclear structure’ , ‘Extracellular structures’ , and ‘Cell motility’  categories represented the smallest groups .To further analyze the chrysanthemum transcriptome, all unigenes were compared with the Kyoto Encyclopedia of Genes and Genomes (KEGG) database using BLASTx with an E-value threshold of < 1e-5. Of the 43,137 unigenes, 13,704 had significant matches in the database and were assigned to 285 KEGG pathways . The mosThe number of unigenes with an RPKM value of > 0.3 that were shared by vegetative buds and floral buds, floral buds and buds, and vegetative buds and buds were 80,769, 81,794, and 80,984, respectively. Vegetative buds, floral buds and buds shared 78,561 unigenes in common . By contThe resulting reads were mapped to our reference transcriptome; statistical significance values were taken to be reliable with an RPKM value ≥ 2 in at least one of the three samples. Transcriptomes were compared, and differentially expressed genes (DEGs) were analyzed in three samples: vegetative buds, floral buds and buds. To determine which of the unigenes were differentially expressed, we filtered with an FDR ≤ 0.05 and a |log2 (ratio)| ≥ 2; 1,876 DEGs were significantly changed in expression between vegetative buds and floral buds, of which 1,516 DEGs were up-regulated and 360 DEGs were down-regulated in the floral buds .GO and KEGG Pathway enrichment analyses were performed on the DEGs to identify differences in the biological processes and pathways between vegetative buds and floral buds. In total, 998 DEGs were enriched in GO categories. A total of 2,983 DEGs  were identified between floral buds and buds . In totaA total of 2,356 DEGs  were identified between buds and vegetative buds . In addi2 = 0.907). By contrast, the difference in gene expression was the largest between floral buds and buds, leading to a poor correlation in gene expression  was greater than the number in buds relative to floral buds (1277). Fewer genes (360) were down-regulated in floral buds relative to vegetative buds than the number down-regulated (1706) in buds relative to floral buds. Indeed, there were only 90 common DEGs among vegetative buds, floral buds and buds . In totaBy comparing the transcriptomes of vegetative buds, floral buds and buds, we found that many unigenes were dramatically up-regulated in floral buds and down-regulated in both vegetative buds and buds, suggesting that these genes play an important role during flower differentiation.CRTCL) that was highly expressed in floral buds, weakly expressed in vegetative buds, and not expressed at all in buds (AP2CL3) was assigned to “organ morphogenesis” (GO:0009887) under the 'biological process' category. Furthermore, many other homologs of these transcription factors were assigned to the ‘nucleus’ (GO:0005634), ‘DNA binding’ (GO:0003677) and ‘regulation of transcription, DNA-dependent’ (GO:0006355) categories under 'cellular component,' 'molecular function' and 'biological process,' respectively.The expression levels of many important transcription factor genes, including the homologs of AP2, MYB, MYC, WRKY, NAC and CRT, were dramatically up-regulated in floral buds . Interes in buds . One homThe genes that were up-regulated during flower differentiation also included many genes that are responsive to phytohormones or are involved in phytohormone synthesis, as well as sugar synthesis/transport genes, kinase-like genes, ubiquitin-like genes, and resistance genes; numerous uncharacterized protein coding genes were also identified. These genes were enriched for the ‘carbohydrate metabolic process’ (GO:0005975), ‘signal transduction’ (GO:0007165), ‘protein kinase activity’ (GO:0004672) and ‘defense response’ (GO:0006952) categories and are listed in CRYPTOCHROME (CRY), PHYTOCHROME (PHY), LATE ELONGATED HYPOCOTYL (LHY), EARLY FLOWERING 4 (EFL4), FLAVIN-BINDING KELCH REPEAT F-BOX 1 (FKF1), TIMING OF CAB 1 (TOC1), ZEITLUPE (ZTL) and GIGANTEA (GI) and others [CRY , six homologs of PHY , one homolog of LHY (LHYCL), one homolog of EFL4 (EFL4CL), four homologs of FKF1 , one homolog of TOC1 (TOC1CL), one homolog of ZTL (ZTLCL), and three homologs of GI . CONSTANS (CO) is a key regulator of the photoperiod response, and one homolog of CO (COCL1) was identified. FT (Flowering Locus T) is the early target of CO, and one homolog of FT (FTCL) was also identified (SUPPRESSOR OF CONSTANS1 (SOC1), SHORT VEGETATIVE PHASE (SVP), LEAFY (LFY), APETALA1 (AP1), GIGANTEA (GI), AGAMOUS (AG) and PISTILLATA (PI). APETALA2 (AP2) also play key roles in promoting floral meristem identity. And homologs of these MADS box genes and AP2 were also identified (LEAFY (LFY) is a vital regulator of the specification of floral meristem identity. This gene is initially expressed very early throughout the presumptive floral meristem, and its expression leads to a cascade of transcriptional activities that control floral meristem development and meristem identity [LFY (LFYCL) that was significantly up-regulated in floral buds. SOC1 is an upstream regulatory gene of LFY. Two homologs of SOC1, SOC1CL1 and SOC1CL2, were identified in chrysanthemum, and their expression levels were significantly up-regulated in the floral bud stage. AP1 is ‘A-class’ like gene; LFY directly activates AP1 transcription [Chrysanthemum is a typical short-day plant; it can flower in response to a single short day. We searched for homologous genes of the important regulators involved in the photoperiod pathway in chrysanthemum. In Arabidopsis, many genes required for the daylength response have been identified using molecular-genetic approaches, among which some members encode regulatory proteins specifically playing roles in the regulation of flowering; others encode components in light signal transduction pathways or pathways related to circadian signaling, including d others . In thisentified . In the entified . LEAFY . In addition, we identified one homolog of the C-class gene AG (AGCL) in chrysanthemum. The homologs of AP3, PI, TM6 and AG, were initially expressed in the floral buds, although they showed the highest expression in buds. Annotation information for unigenes involved in the photoperiod pathway in chrysanthemum is listed in In ignaling . AP1CL1 anthemum , and theirrhinum . In chryMADS-boxCL1, MADS-boxCL2, MADS-boxCL3, GDEF1CL, AP2/ERFCL3, CRTCL, MYBCL1, DoubleWRKYCL, GA2O2CL, ETHR3CL) and 10 were annotated as uncharacterized or predicted proteins . The expression of 4 genes  was only weakly detected in vegetative buds and floral buds by qRT-PCR  were annotated in the Swiss-Prot database, and the remaining 48,230 unigenes  had no Swiss-Prot annotations due to the lack of chrysanthemum genomic and EST information. The assembly unigenes in this study were assigned to an extensive range of GO categories, KEGG pathways and KOG classifications, which indicated that various transcripts are involved in chrysanthemum flower development.CRT/DRECL, CRTCL) showed minimal expression in vegetative buds, and one CRT gene (CRTCL) was not detected in buds. It is known that CRT genes are important regulators of plant reproductive events [Comparative analysis of the transcriptomes revealed a wealth of valuable information. As shown in e events . HoweverCRT/DRECL, SOC1CL1, SOC1CL2, SVP1CL, SVP3CL, AGLCL2, AP1CL1, AP1CL2, AP1CL3, AP2/ERFCL2, AP2CL1, AP2CL2, AP2CL3, AP2/ERFCL1, AP3CL, PICL, TM6CL, AGCL1, AGCL3, and SEPCL) as well as several homologous genes in the ethylene signaling pathway. Another 'biological process' term, ‘regulation of transcription, DNA-dependent’ (GO:0006355), included 37 genes that were the homologs of WRKY, NAC, and CRT , LFY, SOC1, SVP, AGL, AP1, AP2, AP3, PI, TM6, AG, and SEP , as well as genes involved in the ABA and ethylene signaling pathways. The phase transition from vegetative to reproductive growth is a complex biological process that is regulated by a wide range of genes that also interact with one another, leading to a cascade of physiological and biological changes. The functions of certain transcription factors, such as AP2, WRKY, MYB, MYC, NAC and CRT, during the regulation of flower bud differentiation should be further studied in chrysanthemum.Among the important transcription factors mentioned above, some are involved in the same biological processes as genes involved in phytohormone signaling, the photoperiod pathway and organ determination. For example, 27 homologous genes were assigned to ‘transcription, DNA-dependent’ (GO:0006351) in the 'biological process' category. These genes include the homologs of CRT, SOC1, SVP, AGL, AP1, AP2, AP3, PI, TM6, AG, and SEP  a calcium-dependent protein kinase is involved in photoperiodic flower induction in Pharbitis nil. [Genes involved in signal transduction as well as sugar, kinase, ubiquitin and defense pathways were also up-regulated in floral buds relative to vegetative buds and buds. These up-regulated genes were enriched for the GO categories ‘carbohydrate metabolic process,’ ‘protein kinase activity,’ ‘plant hormone signal transduction,’ and ‘defense response,’ among others. These findings indicate that genes involved in carbohydrate metabolic processes, protein kinase activities, plant hormone signal transduction, defense responses and other related processes play a role in floral differentiation in chrysanthemum. This hypothesis is also consistent with previous studies showing that 1) various phytohormones play an important role in the regulation of flowering in both long- and short-day plants; 2) the soluble sugar and starch contents of leaves and buds are temporarily increased in tis nil. , 24, 25.In addition, there were many unigenes coding uncharacterized proteins that are also worthy of attention. Some of these were assigned to flower development-related GO terms, and many others were significantly up-regulated in floral buds, suggesting that they are involved in regulating chrysanthemum flower development. Further studies should be conducted to reveal the exact roles of these uncharacterized genes in the regulation of flower development.CRY, PHY, LHY, FKF1, TOC1, ZTL, GI, CO, FT) encoding components of light signal transduction pathways or related to circadian rhythms. LFY expression leads to a series of transcriptional activities that control floral meristem formation and meristem identity [LFYCL) in chrysanthemum was identified. In addition, the homologs of AP2 and many MADS box genes involved in floral meristem identity were identified in chrysanthemum. According to the ABC model, A function genes  specify the identity of the sepal, A function genes together with B function genes  specify the petals, B function genes together with C function genes (AG) specify the stamen, and C function genes specify the carpel. However, the capitulum of chrysanthemum is composed of hundreds of individual florets which are specialized in structure and function, indicating that the regulatory mechanism of A, B and C function genes should be further studied in this species. The homologous genes of A, B and C function genes were identified in this study. These genes are important candidate genes for molecular cloning and functional analysis of flowering regulation genes in chrysanthemum.The chrysanthemum floral meristem initiates in response to a single short day; the photoperiod is considered to be the most important factor controlling chrysanthemum flowering time. We identified many genes  cultivated in a greenhouse under a 16-hr light/8-hr dark cycle for 180 days and then an 8-hr light/16-hr dark cycle at 23°C in Beijing Forestry University . Under long-day treatment, 70 vegetative buds were collected between 9:00–12:00 a.m. When the short-day treatment began, 70 apical buds were collected between 9:00–12:00 a.m. every week until visible floral buds were formed. After floral buds were formed, 30–50 buds were collected between 9:00–12:00 a.m. every week until the ray florets started to show color. All plant tissues collected were placed immediately in liquid N2 and stored at −70°C until RNA was extracted. The individual plants used for collecting materials were all derived by tissue culture from the C. morifolium genotype ‘Fenditan.’ Total RNA was extracted using an RNAisomate RNA Easyspin Isolation System, and a NanoDrop ND2000 instrument was used to assess the RNA quantity and quality.The tissues  used in the study were obtained from a ground-cover chrysanthemum variety  mRNA was purified using magnetic beads with oligo (dT) and was then fragmented into small pieces. After the first-strand cDNAs were synthesized using random hexamer primers, the second strand was synthesized using appropriate buffers, dNTPs, RNase H, and DNA polymerase I. After purification using the QiaQuick PCR extraction kit, the double-stranded cDNAs were resolved using EB buffer for end reparation and poly (A) addition. We then connected the short fragments with sequencing adapters. Finally, a cDNA library was constructed, which was sequenced using an IlluminaHiSeq 2000. Low-quality reads were removed using an in-house Perl script. The transcriptome sequences were then assembled with clean reads. First, the distinct contigs were assembled using the short reads with the software program CLC Genomic Workbench 5.5. Next, scaffolds between the contigs were constructed by employing the paired-end relationships between the reads. Finally, we filled the intra-scaffold gaps and constructed a non-redundant unigene set from the assembled datasets using CAP3 software .http://www.expasy.ch/sprot), KEGG (http://www.genome.jp/kegg) and KOG (http://www.ncbi.nlm.nih.gov/KOG). The sequence directions of unigenes were identified according to the best alignment results. The Blast2GO program was used to produce GO (http://www.genome.jp/kegg) annotations for the assembled unigenes [For functional annotation, Blastx alignment  was conducted between unigenes and protein databases, including Nr , Swiss-Port  . The traC. morifolium was used as a reference gene [Total RNA was extracted from the vegetative buds, floral buds and buds as described above. After treatment with DNase, the total RNA was subjected to reverse transcription to produce cDNA using a reverse transcription system. Real-time RT-PCR was conducted using the PikoReal real-time PCR system. Each reaction was performed in a total reaction mixture volume of 20 μL, which contained 2 μL of first-strand cDNA as template. The amplification program was: 30 s at 95°C, 40 cycles of 5 s at 95°C and 30 s at 60°C. Gene-specific primers  were desnce gene .S1 Table(XLSX)Click here for additional data file.S2 Table(XLSX)Click here for additional data file.S3 Table(XLSX)Click here for additional data file.S4 Table(XLSX)Click here for additional data file.S5 Table(XLSX)Click here for additional data file.S6 Table(XLSX)Click here for additional data file.S7 Table(XLSX)Click here for additional data file.S8 Table(XLSX)Click here for additional data file.S9 Table(XLSX)Click here for additional data file.S10 Table(XLSX)Click here for additional data file.S11 Table(XLSX)Click here for additional data file.S12 Table(DOCX)Click here for additional data file.


Hence, there is an urgent need to develop novel approaches to overcome this limitation.The recent use of T cell-based cancer immunotherapies, such as adoptive T-cell transfer and checkpoint blockade, yields increasing clinical benefit to patients with different cancer types. However, decrease of MHC class I expression is a common mechanism transformed cells take advantage of to evade CD8NLRC5 has been recently described as a key transcriptional regulator controlling expression of MHC class I molecules. In this commentary, we summarize and put into perspective a study by Rodriguez and colleagues recently published in Oncoimmunology, addressing the role of NLRC5 in melanoma. The authors demonstrate that NLRC5 overexpression in B16 melanoma allows to recover MHC class I expression, rising tumor immunogenicity and counteracting immune evasion. Possible ways of manipulating NLRC5 activity in tumors will be discussed. Highlighting the therapeutic potential of modulating NLRC5 levels, this publication also encourages evaluation of NLRC5, and by extension MHC class I pathway, as clinical biomarker to select personalized immunotherapeutic strategies. Loss or down-regulation of MHC class I expression, accompanied by impaired expression of one or several components of the antigen processing machinery (APM), is frequently observed in many solid and hematopoietic tumors . Th. Th10]. reatment , 16–18, reatment , 20. It reatment .Nlrc5-knockout mouse models are valuable tools to address these points, in addition to a thorough analysis of NLRC5 transcript levels and somatic mutations in human tumors. Indeed, data reporting NLRC5 alterations are starting to emerge [Only recently, with the advent of the so-called “checkpoint inhibitor” and adoptive T cell transfer therapies, we fully appreciate the power of the antitumoral cytotoxic immune response , 22. Howo emerge –26, showAPM, antigen processing machinery; CTLA-4, cytotoxic T-lymphocyte-associated protein 4; IFN, interferon; MHC, major histocompatibility complex; NK, natural killer; NLRC5, NLR caspase recruitment domain containing protein 5; NLR, NOD-like receptor; NOD, nucleotide-binding oligomerization domain; TA, tumor antigen; TCR, T cell receptor


Human papillomavirus (HPV) types 16 and 18 cause invasive cervical cancer and most invasive anal cancers (IACs). Overall, IAC rates are highest among men who have sex with men (MSM), especially MSM with HIV infection. Testosterone is prescribed for men showing hypogonadism and HIV-related wasting. While there are direct and indirect physiological effects of testosterone in males, its role in anal HPV16/18 infections in men is unknown.Free testosterone (FT) was measured in serum from 340 Multicenter AIDS Cohort Study (MACS) participants who were tested for anal HPV16/18-DNA approximately 36 months later. The effect of log10-transformed current FT level on anal HPV16/18 prevalence was modeled using Poisson regression with robust error variance. Multivariate models controlled for other HPV types, cumulative years of exogenous testosterone use, race, age, lifetime number of receptive anal intercourse partnerships, body mass index, tobacco smoking, HIV-infection and CD4+ T-cell counts among HIV-infected, and blood draw timing.Participants were, on average, 60 (+5.4) years of age, White (86%), and HIV-uninfected (56%); Twenty-four percent tested positive for anal HPV16 and/or 18-DNA . In adjusted analysis, each half-log10 increase of FT was associated with a 1.9-fold  higher HPV16/18 prevalence. Additionally, other Group 1 high-risk HPVs were associated with a 1.56-fold  higher HPV16/18 prevalence. Traditional risk factors for HPV16/18 infection  were poorly correlated with one another and not statistically significantly associated with higher prevalence of HPV16/18 infection in unadjusted and adjusted analyses.Higher free testosterone was associated with increased HPV16/18 prevalence measured approximately three years later, independent of sexual behavior and other potential confounders. The mechanisms underlying this association remain unclear and warrant further study. Gay, bisexual and other men who have sex with men (MSM) are at high risk for both human immunodeficiency virus (HIV) and human papillomavirus (HPV) infections –3. Over Sex hormones have been linked to HPV-related cervical cancers through the association between oral contraceptive (OC) use and parity in women . For exaAvailable data suggest that frailty, hypogonadism, and wasting disease symptoms; as well as age and obesity are associated with lower circulating serum testosterone levels –25. In HThe Multicenter AIDS Cohort Study (MACS) protocol and its substudies, including the Anal Health Study (AHS), were approved locally by institutional review boards at each of four U.S. MACS study sites:  Johns Hopkins Bloomberg School of Public Health, (Chicago) Northwestern University, (Los Angeles) UCLA, and (Pittsburgh) University of Pittsburgh. Written informed consent for study procedures, specimen collection and testing were provided by the sample of 340 men evaluated for this study.Overall, the MACS has enrolled 6,972 HIV-infected and -uninfected MSM in its lifetime: 4,954 men were enrolled between 1984–1985, 668 men between 1987–1991, and 1,350 men between 2001–03 1,350 , 28. By These analyses reflect data from two separately conducted MACS sub-studies that evaluated serum testosterone and anal HPVs among HIV-infected and -uninfected men. The testosterone study (parent study) evaluated the hypothesis that HIV-infection affected serum FT levels using a nested case-control study design. Specifically, serum specimens for 542 HIV-infected and -uninfected men ≥45 years of age were retrospectively selected from the MACS repository and tested for testosterone. Peripheral blood specimens collected from all men evaluated at each study visit were separated into cells and serum at local sites and aliquots of varying volumes were cryopreserved for long-term storage. Frozen serum specimens were shipped monthly on dry ice to the national MACS repository for all men completing a study visit. For these analyses, testosterone-tested specimens for the parent study had been frozen and thawed once, at most, prior to (testosterone) testing. Bolelli, et al. and others suggest testosterone analytes are stable in frozen serum 10 or more years –31 and tCase and control specimens tested for the parent study were selected from repository samples and up to six serum specimens/subject were evaluated. Controls were matched to cases based upon gender, race, enrollment period, study site, and specimen-collection  time, and age (±5 years). HIV-infected men (cases) were selected if ≥3 specimens were available, with ≥1 samples collected ≤3 years before beginning highly active antiretroviral therapy (HAART), and ≥2 samples collected within 10 years following HAART initiation. The availability and initiation of HAART varied among cases, and accordingly, temporally-matched test specimens ranged widely. For men who later enrolled in the Anal Health Study, first specimens tested for testosterone were collected between May, 1993 and February, 2006; similarly, the last specimens tested in each subject’s specimen set were collected between June 1997 and January 2010. For each man, testosterone data for only one visit were included in the analysis (as described in the “exposure of interest” section below).HPV data were collected as part of a second sub-study that began active enrollment of 1,504 men in March 2010. Anal swab specimens were collected using a standardized clinical protocol and all active MACS men were eligible to participate . DNA extThus, this study is best conceptualized as a voluntary follow-up study of 340 HIV-infected and—uninfected men who were tested for FT as part of a passive, nested case-control analysis of associations between serum FT, HIV-infection and HAART . Specifioutcome of interest was HPV Type 16 or 18 DNA detected in anal swab specimens. Each specimen was collected using a standard clinical protocol: A Dacron swab was inserted approximately 2 inches beyond the anal verge, approximated to the wall and withdrawn, while rotated, over ~30 seconds before being placed into 20 mL of preservative  [The ugh, MA) . A vorteugh, MA) . Denaturugh, MA) , 35, 36.exposure of interest, was derived from total testosterone measured in cryopreserved serum using liquid chromatography-tandem mass spectrometry, and sex-hormone binding globulin estimated from radioimmunoassay [10-transformed data, β = -0.003 (+0.06) and β = -0.007 (+0.03), respectively. FT measured in serum was not statistically significantly associated with the time lapse between the last FT measurement and the first HPV test visit . Thus, the FT measurement closest to the HPV test visit was selected and evaluated for these analyses.FT, the unoassay . The Verunoassay . For thiunoassay , 39. The2, BMI), HIV-infection characteristics at the HPV test visit, and Hepatitis C virus (HCV) infection, known to be associated with FT [p-values<0.001); for this reason, only smoking closest to the HPV test visit was included in the final model. Those with BMI ≥25 kg/m2 were compared to subjects with BMI <25 kg/m2 men. For 32 men missing BMI measures at the testosterone visit, BMI measured at the closest MACS visit was used. Men reporting >500, 201–500, and 51–200 lifetime sex partners at MACS Visit 1 were compared to men who reported ≤50 sex partners at MACS Visit 1. Men who reported ≥355, 133–354, and 63–132 partners, representing quartile cutoffs, between MACS Visit 1 and the visit 24 months before HPV testing were compared to men who reported <63 partners over the same interval. Men who reported ≥4 and 1–3 receptive anal intercourse (RAI) partners for the last 24 months prior to HPV testing were compared to those who reported none. For 5 men missing RAI partnership data for ≥1 study visits during the last 24 months of observation, the number reported at each of the last four visits with complete RAI data before HPV testing were included. Lifetime cumulative duration of exogenous testosterone use in years was calculated for men who reported ever using exogenous testosterone. For these analyses, exogenous testosterone products included Androderm , AndroGel , Delatestryl , Striant , Testoderm , and Virilon . Data for three MACS study sites were compared to data from the site with the largest population, and men who enrolled early in the MACS, before 2001, were compared to those enrolled in 2001–2003 to control for sociodemographic differences. Time of phlebotomy was included in the analysis to control for differences associated with diurnal variation in testosterone secretion: before vs. after noon.We evaluated demographic characteristics that were collected at semiannual visits, including age, race (White non-Hispanic vs. others); other Group 1, Group 2 and low risk HPVs; tobacco smoking, alcohol use, body mass index  and 95% confidence intervals (95% CI) are reported . The eff+): 5.4) years of age, White , and more than half were HIV-uninfected  pg/mL), especially among HIV-infected men: 82.9 (+78.3) vs. 70.9 (+21.2), for HIV-uninfected men  (p = 0.04)  , and HIV = 0.04)  differed3 had a higher prevalence of HPV16/18 infection than HIV-infected men with >500 cells/mm3 and HIV-uninfected men: 37% (23/63) vs. 24% (21/88) vs. 19% (36/189), respectively . Additionally, the mean and variation of FT levels for HPV16/18-infected men was greater than that for men that tested positive for other Group 1 hrHPVs and those who tested negative for all Group 1 viruses (Nearly 24% (80/340) of participants tested HPV16/18-positive; of these, 17% (58/340) tested positive for HPV16 and 9% (31/340) showed HPV18-DNA. Only 41% (33/80) tested positive for HPV16 or-18 alone from among the 12 Group 1 hrHPVs [ viruses ..10 increase in FT ), even after controlling for age, race, cumulative years of exogenous testosterone use, self-reported lifetime number of sexual partnerships, BMI, tobacco smoking, HIV-infection and CD4+ T-cell count, among the HIV-infected men, and the timing of blood draws  and 1.23 , respectively), nor did partnerships reported earlier in the study period  may not adequately control for fluctuations in testosterone associated with physiologic diurnal variation. Also, other sex hormones were not measured; for example, we cannot evaluate the effect that testosterone conversion to estradiol, through the aromatase pathway, might have on HPV16/18 prevalence in these men . NonetheIdeal use of testosterone-replacement therapy returns testosterone levels to age-appropriate norms, relieves symptoms, and prevents disease for deficient men without harmful side-effects , 60, 61.S1 Table(DOCX)Click here for additional data file.S1 FigThree Hundred and Forty Men Participated in the Anal Health Study for HPV Genotyping Nearly Three Years Later.(TIF)Click here for additional data file.S2 Fig(TIF)Click here for additional data file.S3 Fig(TIF)Click here for additional data file.


The genetic mechanisms underlying hemangioblastoma development are still largely unknown. We used high-resolution single nucleotide polymorphism microarrays and droplet digital PCR analysis to detect copy number variations (CNVs) in total of 45 hemangioblastoma tumors.EGFR and PRDM16 genes. Recurrent losses were located at chromosome 12 (q24.13), which includes the gene PTPN11.We identified 94 CNVs with a median of 18 CNVs per sample. The most frequently gained regions were on chromosomes 1 (p36.32) and 7 (p11.2). These regions contain the EGFR, PRDM16, PTPN11, HOXD11, HOXD13, FLT3, PTCH, FGFR1, FOXP1, GPC3, HOXC13, HOXC11, MKL1, CHEK2, IRF4, GPHN, IKZF1, RB1, HOXA9, and micro RNA, such as hsa-mir-196a-2 for hemangioblastoma pathogenesis. Furthermore, our data implicate that cell proliferation and angiogenesis promoting pathways may be involved in the molecular pathogenesis of hemangioblastoma.Our findings provide the first high-resolution genome-wide view of chromosomal changes in hemangioblastoma and identify 23 candidate genes:  Hemangioblastomas (HB) are highly vascular tumors, which account for approximately 3 % of all tumors of the central nervous system (CNS) . It occu(VHL) tumor-suppressor gene. The VHL protein, which is the critical part of a ubiquitin ligase protein complex that binds to the hypoxia-inducing factors HIF-1 and HIF-2 transcription factors and targets them for ubiquitination and proteosomal degradation. Dysregulation of this VHL-associated function causes increased expression of a variety of growth factors, including erythropoietin, PDGF, VEGF and TGF. Upregulation of these factors may lead to angiogenesis and tumorigenesis. Additional mechanisms of tumorigenesis have been described outside of the HIF pathway, including alterations in microtubule binding and stabilization, abnormal extracellular matrix composition as well as apoptosis and transcription regulation . Am. AmFLT3)d tumors . On the d tumors . miR-196d tumors . The miRarcinoma . Accordi the CNS . Prdm16 ntenance . Megakaransition . Moreoveeoplasms . Finallyntiation . The faca (JMML) , acute ma (JMML) , and acua (JMML) . Overexpa (JMML) . In contin liver  and cartin liver . Accordipecimens . Thus, iFOXP1) and p25 (3 samples showed PPARG loss and 2 samples showed VHL loss). Interestingly, FOXP1 transcription factor, located on chromosome 3(p13), can function as a tumor suppressor gene. Low expression in glioma [Previous analysis of six VHL-related CNS hemangioblastomas showed loss of chromosome 3p or the whole of chromosome 3 to be the most common abnormality, which is detected in 70 % and loss of 1p11-p31 in 10 % . More ren glioma  and Hodgn glioma . DiffereChek2 is deleted and EGFR, PTPN11, Ptch1 amplified in majority of hemangioblastoma patients. EGFR is the only gene that has been previously reported as a candidate gene with hemangioblastoma. Independent of HB tumor location PTPN11 may act as tumor suppressor or oncogene depending on the tumor cell of origin. These findings have potentially relevant clinical value, as this the first high resolution for chromosomal alteration in HB. Future research should be dedicated to the prospective validation of these alterations and further characterization of tumors that carry the deletions/amplifications, as well as of defining the role of these genes. This may offer insights into hemangioblastoma biology, provide DNA-based markers that can be analyzed by FISH suitable for routine clinical applications and eventually lead to the development of effective targeted therapies for HB.In this study, we have demonstrated in two different tumor cohorts and using two different techniques for copy number alteration detection, SNP and digital PCR, that


After viral infection, host cells respond by mounting an anti-viral stress response in order to create a hostile atmosphere for viral replication, leading to the shut-off of mRNA translation (protein synthesis) and the assembly of RNA granules. Two of these RNA granules have been well characterized in yeast and mammalian cells, stress granules (SGs), which are translationally silent sites of RNA triage and processing bodies (PBs), which are involved in mRNA degradation. This review discusses the role of these RNA granules in the evasion of anti-viral stress responses through virus-induced remodeling of cellular ribonucleoproteins (RNPs). The control of the translation and turnover of mRNAs plays fundamental roles in the regulation of gene expression. Eukaryotic cells have developed different mechanisms to control these processes in order to respond to environmental stress, such as heat shock, UV irradiation, hypoxia, endoplasmic reticulum (ER) stress and viral infection. Among those mechanisms is the assembly of non-membrane-delimited bodies, called RNA granules, that contain RNA-binding proteins (RBPs) and translationally silenced mRNAs ,2.The classification of RNA granules is based on a number of factors, including their subcellular localization , the presence of specific markers, the cell type where they are assembled , response to stimuli , dynamic behavior and proposed functions  . The nucSGs and PBs are associated with mRNA storage and decay, respectively . ImportaSGs are composed of target mRNAs bound to translation initiation factors  to allow the rapid restoration of translation once the stress is gone ,28. ManyPBs contain components of the mRNA decapping machinery  ,41, scafSeveral viruses modulate the assembly of SGs to promote viral replication and suppress the cellular stress response. The discussion below was performed upon the interaction between virus families and SGs .Herpesviridae and Poxviridae are enveloped double-stranded DNA (dsDNA) viruses. Herpes simplex virus type 1 (HSV-1) shuts off host protein synthesis by impairing the activation of eIF2 kinases through the virion host shutoff (vhs) protein [Members of the family  protein , Us11 [1 protein , ICP34.5 protein  and glyc protein . During  protein ,134, rel protein . Recentl protein . The blo protein . Human c protein , while s protein .Poxviridae family, replicates within the cytoplasm in large foci called DNA factories that co-opt SG proteins, such as G3BP1, Caprin1, eIF4E, PABP and eIF4G [Vaccinia virus (VV), a member of nd eIF4G ,141. VV nd eIF4G ; viral tnd eIF4G , suggestnd eIF4G ; howevernd eIF4G .Reoviridae is composed of non-enveloped virions with a 9–12 dsRNA segment genome. The prototypical member of the family, rotavirus, causes the shut off of host protein synthesis [The family ynthesis , and desynthesis . The perynthesis . By contynthesis . Howeverynthesis . Recentlynthesis .Picornaviridae family are composed of non-enveloped particles. Evidence indicates that poliovirus (PV) proteinase 2A induces assembly of SGs early post-infection (between 2 and 4 h) [All members of the and 4 h) ,149, whiand 4 h) . Interesand 4 h) ,152. Encand 4 h) ,153. By and 4 h) ,155. A mand 4 h) .Togaviridae family composed of enveloped virions, is able to induce eIF2α phosphorylation promoting SG assembly at early stages of infection [In 2005, McInerney and colleagues showed that Semliki Forest virus (SFV), a member of the nfection . Howevernfection . It was nfection . Chikungnfection . Later, nfection , colocalnfection . G3BP2-cnfection . In contnfection . Finallynfection ,164 and nfection . In addinfection .Flaviviridae family are composed of enveloped virions and West Nile virus (WNV) was the first virus reported to block SG assembly. Li and colleagues showed that the 3` stem loop in the viral genome is able to capture TIA-1 and TIAR [Flaviviridae pestivirus, impairs the arsenite-mediated SG assembly; despite the fact that BVDV N-terminal protease (Npro) is able to interact with several RNA granules components, such as YB-1, IGFBP2, DDX3, ILF2 and RHA (DXH9) [The and TIAR . In addiand TIAR . Howeverand TIAR . In anotand TIAR . In addiand TIAR , and it and TIAR . Consistand TIAR . On the and TIAR . There iA (DXH9) . ContrasA (DXH9) . In addiA (DXH9) . They reA (DXH9) . RecentlA (DXH9) . HoweverA (DXH9) ,180. TheA (DXH9)  and how A (DXH9) ,181,182.Dicistroviridae family, prevents the aggregation of Rox8 and Rin, Drosophila SG marker homologs of TIA-1 and G3BP-1, respectively, even in the presence of various stressor, such as arsenite, pateamine A or heat shock [The infection with Cricket paralysis virus (CrPV), a member of at shock . Althougat shock Coronaviridae is composed of membrane-enveloped virions and one of the most significant features of coronaviruses is the expression of downstream genes via transcription of multiple 3` nested subgenomic mRNAs [The family ic mRNAs . It has ic mRNAs ,186. MHVic mRNAs , while TCaliciviridae family is composed of non-enveloped virions and recently, Humoud and colleagues showed that the feline calicivirus (FCV) infection triggers eIF2α phosphorylation without inducing SG assembly [Finally, assembly . This reassembly . On the assembly , but doeassembly Orthomyxoviridae is composed of enveloped virions with a segmented, negative ssRNA genome [The family of A genome . InfluenA genome . The abiA genome  or by exA genome . The NS1A genome . BesidesA genome . In addiA genome ), ThulasA genome .Arenaviridae, a family of enveloped virions with a bi-segmented negative-sense single strand RNA ((-)ssRNA) genome, that prevents SG assembly by impairing the phosphorylation of eIF2α [Junin virus (JUNV) is a virus belonging to of eIF2α , while tof eIF2α . In addiof eIF2α , suggestRhabdoviridae, induces the phosphorylation of eIF2α and promotes the assembly of SG-like particles that contain TIA-1, TIAR, PCBP2, viral replication proteins and RNA, but not the eukaryotic initiation factor 3 (eIF3), nor eIF4A [Vesicular stomatitis virus (VSV), a virus in the family or eIF4A , suggestParamyxoviridae family are composed of enveloped virions. Respiratory syncytial virus (RSV) induces SG assembly mediated by PKR, and its formation enhanced RSV replication [Members of lication ,197. Givlication  as well lication . Moreovelication . Measleslication . Intereslication . Howeverlication . Sendai lication . In addilication . This stlication )Bunyaviridae are a large family composed of enveloped virions with a tri-segmented (-)ssRNA genome [A genome . The pecA genome . These mA genome ,204. ThiA genome . AnotherA genome . AlthougA genome ,208. MorA genome ,210. TheOn the other hand, Bunyaviral proteins, like other viral proteins, have evolved mechanisms to inactivate PKR and inhibit the IFN response. Similarly to the non-structural protein NS1 from IAV  or NS4A Filoviridae family composed of enveloped virions, does not trigger eIF2α phosphorylation or SG formation. However, SG components are sequestered within viral inclusions where they colocalize with viral mRNA [Recently, Nelson and colleagues showed that the infection with Ebola virus(EBOV), member of ral mRNA . In addiral mRNA .Retroviruses are enveloped, positive ssRNA viruses, which synthesize complementary DNA (cDNA) by reverse transcription that is integrated into the host chromosomal DNA . The oncSeveral viruses are known to disrupt P-bodies. Here, we will summarize the most up-to-date information known about the relationships between viruses and PBs . Adenovirus regulates its gene expression over the time of infection, triggering an accumulation of viral late mRNA in the cytoplasm . To prevA recent report showed that rotavirus infection triggers a significant time-dependent decline of PBs components XRN1, DCP1, Pan3, but not GW182 . Pan3, bIn the case of flaviviruses, WNV and DENV infections reveal a reduction in PB formation . Indeed,On the other hand, infection with picornaviruses, such as PV and CVB3, entirely disrupts PB formation through the virally-induced cleavage of Dcp1a mediated by viral protease 3C, as well as the degradation of Xrn1 and Pan3 mediated by the proteasome . It has The negative-strand RNA viruses are known to use cap-snatching mechanism, to initiate its own mRNA synthesis. However, for IAV, this process occurs in the nucleus, while that for Bunyavirus occurs in PBs. Mir and colleagues showed that hantavirus nucleocapsid protein (N) avoids the 5′ cap degradation of cellular mRNAs, protecting them from Dcp1a/Dcp2-mediated decapping, . In addiLastly, the protein APOBEC3  provides anti-HIV-1 activity while being a PB component that interacts with Ago2, Dcp1a, Dcp2 and DDX6 ,115. HowAlthough significant advances have been made to understand how viruses modulate the assembly/disassembly of RNA granules, there are still outstanding questions that need to be addressed. For example, could these mechanisms be targets of new antiviral drugs? What are the molecular mechanisms and/or signaling pathways that transport mRNAs from one RNA granule to another? Do post-translational modifications that serve as signals to modulate the formation of RNA granules exist? To answer these questions, several reports have helped us to comprehend the molecular biology of RNA granules ,269,270,


Rheumatoid arthritis is an autoimmune arthritis in which two inflammatory cytokines, tumor necrosis factor-α and interleukin-1β, play a critical role in the induction and progression of the disease. Several reports and data from registries have discussed the association between chronic inflammatory diseases and disorders in intermediary metabolism, pointing out that prevalence of peripheral insulin resistance and type 2 diabetes mellitus is increased among patients with rheumatoid arthritis. In addition, several studies have shown that type 2 diabetes mellitus may be considered an interleukin-1β inflammatory-mediated process, and both preclinical and clinical observations have reported the usefulness of interleukin-1 antagonism therapy in this disease.We describe the case of a 58-year-old Caucasian woman and a 74-year-old Caucasian man with rheumatoid arthritis associated with type 2 diabetes mellitus. In these patients, the inhibition of interleukin-1β not only induced remission for rheumatoid arthritis, but successfully controlled their metabolic status.We report the positive effects of the inhibition of interleukin-1 in two patients with rheumatoid arthritis associated with type 2 diabetes mellitus, with both reaching the therapeutic targets of their diseases by using a single biological agent and tapering or discontinuing their antidiabetic therapies. These findings suggest that targeting interleukin-1 might be considered a good therapeutic option for the treatment of rheumatoid arthritis associated with type 2 diabetes mellitus. Interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) play a critical role in the induction and progression of rheumatoid arthritis (RA), and the efficacy of therapies targeting these two inflammatory cytokines confirms their prominent role in the disease ,3. In adA 58-year-old Caucasian woman was referred to our clinic due to the insidious onset, in previous months, of arthralgias and arthritis. Her symptoms involved wrists, hands, shoulders and knees. She also reported notable morning stiffness and fatigue, limiting her daily activities. The results of her cardiological, pulmonary, dermatological, abdominal and neurological examinations were unremarkable. In her medical history, both essential hypertension and osteoporosis were reported. Laboratory findings showed an increase of inflammatory markers and were negative for rheumatoid factor (RF), antinuclear antibodies and anti-cyclic citrullinated peptide (ACPA). A radiological examination of her wrists and hands showed multiple bone erosions and juxta-articular osteoporosis. Her Disease Activity Score in 28 Joints (DAS28), Simple Disease Activity Index (SDAI) score and Patient Global Visual Analogue Scale (PG-VAS) score were 4.5, 28 and 67mm, respectively, and active seronegative RA was diagnosed. Combination therapy with methotrexate (15mg/weekly) , hydroxychloroquine (400mg/daily)  and a low dose of prednisone (10mg/daily)  was prescribed. She experienced a long clinical remission .She experienced a new disease flare-up after two years, characterized by multiple arthritis  and an increase of morning stiffness and fatigue . Infliximab (400mg/bi-monthly) was introduced into her treatment regimen and a new clinical remission was reached  for seven years. At this time, after her diagnosis of T2DM (fasting plasma glucose (FPG) level 189mg/dL, glycated hemoglobin (HbA1c) level 62mmol/mol, 7.8% (4.8-5.9%)), she began taking a new oral hypoglycemic drug (metformin 500mg trice/daily). One year later, following a new flare-up of her disease involving arthritis of her wrists, hands, elbows, shoulders and knees , her anti-TNF-α treatment with infliximab  was discontinued. Both her methotrexate and steroids dosage remained stable; the hydrossicloroquine was discontinued due to poor compliance. Anakinra , a recombinant form of a human IL-1 receptor antagonist, was introduced into her therapy regimen (100mg/daily).With regards to therapy for her T2DM, a stable dosage of metformin was continued . In the six subsequent months a new clinical remission was observed. After three months of therapy, her DAS28, SDAI and PG-VAS scores were 3.88, 24.2 and 74mm, respectively. At the same time, her FPG and HbA1c levels were 108mg/dL and 46mmol/mol, 6.3%, respectively. After six months of therapy, her DAS28, SDAI and PG-VAS scores were 2.52, 9.2 and 30mm, respectively. Furthermore, repeated tests showed a stable further reduction in her FPG and HbA1c levels at 103mg/dL and 46mmol/mol (6.3%) (4.8-5.9%), respectively. In Figure A 74-year-old Caucasian man with an eight-year history of RA, treated by a combination therapy with methotrexate (15mg/weekly), hydroxychloroquine (200mg/daily) and methylprednisolone  , obtaining a stable and sustained good clinical response (DAS28 not available) was admitted to our unit with a new flare-up of the disease consisting of multiple swelling joints, mainly involving his hands and feet, with a symmetrical pattern. He showed DAS28, SDAI and PG-VAS scores of 3.78, 11.5 and 51mm, respectively. In addition, screening for autoimmunity highlighted positivity for RF (100UI/mL) and ACPA (1326UI/mL). He also reported a history of uncontrolled T2DM in the last 12 months, treated with repaglinide 0.5mg twice daily  ) despite the lower levels of methylprednisolone (3 to 4mg/daily)  employed. Anakinra (100mg/daily subcutaneously)  was started, in addition to methotrexate (10mg/weekly)  and methylprednisolone (3-4mg daily) , leading to clinical improvement within a few days.At his three-month follow-up, his DAS28, SDAI and PG-VAS scores had dropped to 1.53, 3.3 and 22mm, respectively. In addition, although oral anti-diabetic therapy was tapered off eight days after anakinra introduction due to a poor compliance, an improvement in glycemic control was noted , respectively). At six months his DAS28, SDAI and PG-VAS scores were 1.89, 6.7 and 20mm, respectively, and a further improvement of both his FPG and HbA1c levels was observed , respectively). His fasting insulin level was increased following treatment with anakinra: 36pmoles/liter at baseline, 53pmoles/liter at three months and 77pmoles/liter at six months, respectively. Similarly, we observed that his fasting C-peptide levels were increased: 0.6nmoles/liter at baseline, 1.1nmoles/liter at three months and 1.5nmoles/liter at six months, respectively. His weight, frequency of physical exercise and diet/caloric intake did not modify during the follow-up. No side effects were observed.We show that anakinra, prescribed for the treatment of relapsing RA in two rheumatic patients with associated T2DM, controls not only the clinical pictures of RA, but also their metabolic status, confirming previous reports suggesting that IL-1 blockade may be helpful in patients with T2DM and inflammatory rheumatic diseases .In both case studies, our patients reached a sustained DAS28 remission after introduction of anakinra, without any side effects observed during the follow-up periods. IL-1β is highly expressed in RA, promoting activation of leukocytes, endothelial cells, chondrocytes and osteoclasts [Diabetes mellitus is a chronic disease due to a failure in both producing and using insulin. When compared with healthy people, patients with T2DM display a two-four-fold increased risk of coronary artery disease . On the In our patients, we observed that anakinra administered to treat RA controlled their metabolic disorder. A significant reduction of both FPG and HbA1c levels was observed. It is well known that the pharmacologic interventions for T2DM are aimed at improving both FPG and HbA1c levels, as suggested by the American Diabetes Association and the American Association of Clinical Endocrinologists . Large cIt must be pointed out that when diabetic patients are treated with oral antidiabetic drug monotherapy, after an initial improvement, a progressive and inexorable decline in β-cell function may be observed. Many studies showed that after three years of treatment, the disease may be controlled by monotherapy in only 50%, and in only 25% after nine years , confirmTaken together, these results support that IL-1 antagonism may be a specific therapy in RA with associated T2DM. Although the role of IL-1β has been extensively studied in RA, recently several studies have suggested this cytokine as a key cytokine in the pathogenesis of T2DM ,5. In faet al. performed a double blind, parallel-group trial, involving 70 T2DM patients randomly assigned to receive 100mg of anakinra or placebo. At 13 weeks, the anakinra group showed an improvement of HbA1c levels, the C-peptide secretion was enhanced and a reduction in the ratio of proinsulin to insulin and in levels of IL-6 and C-reactive protein (CRP) was observed [et al., and suggesting an improvement of β-cell function. In this context, it has been recently shown that anakinra is able to increase insulin secretion [In this setting, Larsen ecretion , and thiet al. reported that anakinra therapy was not effective in type 1 diabetes (T1DM) patients [Recently, Moran patients . It mustpatients ,15.In conclusion, we report the positive effects of anakinra for two patients with RA associated with T2DM reaching the therapeutic targets of both the diseases, using a single biological agent, and allowing us to taper or discontinue their antidiabetic therapies. In this setting, further specifically designed studies must be performed to evaluate the safety and long-term efficacy of anakinra in patients with RA associated with T2DM, and the subsequent reduction of CV events in this population. On these bases, an open, randomized, controlled, double-arm, multi-center study, whose primary endpoint is the efficacy of anakinra in controlling the signs and symptoms of T2DM in patients with RA affected by this metabolic disorder, is actually ongoing in our country .Written informed consent was obtained from the patients for publication of this case report. A copy of the written consent is available for review by the Editor-in-Chief of this journal.


Drosophila, Xenopus, and mouse. However, I was confident that WNT signaling should be investigated for clinical application by using human samples or cell lines. In 1998, I, together with post-doctoral fellows, started a human WNTome project to comprehensively clone and characterize novel human genes encoding WNT signaling molecules and to establish a “human WNT research” platform , WNT9B (WNT14B), and WNT10A as the major products of the human WNTome project [reviewed in Katoh , ANO2 (TMEM16B), ANO3 (TMEM16C), ANO4 (TMEM16D), ANO5 (TMEM16E), ANO6 (TMEM16F), ANO7 (TMEM16G), ANO8 (TMEM16H), ASXL2, ASXL3, BCL9L, CDC50A (TMEM30A), CDC50B (TMEM30B), CDC50C (TMEM30C), CRB2, DACT1 (DAPPER1), DACT2 (DAPPER2), DIXDC1, FAT4, FMNL1, FMNL2, FMNL3, FOXR1 (FOXN5), FOXR2 (FOXN6), HES2, HES3, HES5, JMJD1C (TRIP8), JMJD2A (KDM4A), JMJD2B (KDM4B), JMJD2C (KDM4C), JMJD2D (KDM4D), KIF27, MPP7, PRICKLE1, and PRICKLE2.Most human genes that encode WNT signaling components had been cloned and characterized by 2002, whereas thousands of novel human genes outside of the WNT field still remained to be discovered. In 2003, colleagues and I started a post-WNTome project to identify and characterize novel human genes encoding adhesion molecules, transmembrane proteins, epigenetic regulators, and transcription factors Figure . My grouThe human WNTome and post-WNTome projects were gene-hunting adventures that utilized molecular biology and computational biology, respectively. Inter- and intra-cellular signaling networks were simplified to a secondary picture consisting of nodes and edges. Nodes correspond to genes, mRNAs, proteins, or micro-RNAs (miRNAs), while edges correspond to their interactions. I then shifted my interest from the nodes to the edges and the whole picture. In 2007, my laboratory started a stem-cell signaling network project to elucidate mutual interactions of the WNT, FGF, Notch, Hedgehog, TGF-β, and BMP signaling cascades Figure  Katoh a.Recently, I was appointed as the chief editor of Frontiers in Molecular Medicine, a sub- or specialty journal of Frontiers in Cell and Developmental Biology. I would like to contribute to the global scientific community through Frontiers in Molecular Medicine, which aims to address the gap between cell and developmental biology and clinical medicine and to promote development of novel diagnostics and therapeutics for a variety of human diseases, including cancers, cardiovascular diseases, diabetes mellitus, eye diseases, inflammatory bowel diseases, kidney diseases, liver diseases, neurological diseases, and respiratory diseases.Wnt1, Wnt3, Wnt10b, Fgf3, Fgf4, or Fgf10 loci and induces mammary tumorigenesis [Dickson et al., Mouse mammary tumor virus (MMTV) integrates at the Receptor tyrosine kinases (RTKs) are single-span transmembrane receptors with extracellular ligand-binding domain and intracelullar tyrosine kinase domain and are involved in growth factor signaling to downstream cascades, including the RAS-MAPK, PI3K-AKT, SRC, and PKC signaling cascades. FGFR1, FGFR2, FGFR3, and FGFR4 are receptors of the FGF family of ligands  are seven transmembrane receptors that are linked to small G-protein signaling and other atypical signaling cascades. WNT receptors, including Frizzled-1 (FZD1), FZD2, FZD3, FZD4, FZD5, FZD6, FZD7, FZD8, FZD9, and FZD10, as well as Hedgehog signal transducer Smoothened, belong to the GPCR superfamily  gene within a cancer-associated deleted region in human chromosome 11q23.3 in 2004  family members are DNA-binding proteins with a FOX domain that consists of two wing-like loops and three α-helices  repressor complex and the trithorax-group (trxG) activator complex , NCOA1, the retinoic acid receptors (RARα and RARβ), estrogen receptor (ER), and glucocorticoid receptor (GR) are binding partners of ASXL1. Germ-line mutations in ASXL1 occur in Bohring-Opitz syndrome, while somatic mutations in ASXL1 occur in colorectal cancer with microsatellite instability, hematological malignancies and castration-resistant prostate cancer (Gelsi-Boyer et al., The Jürgens, . ASXL1 (Jürgens, , ASXL2 (Jürgens, , and ASXJürgens, , which cExtracellular DNA and circulating miRNAs are key topics in translational medicine (Wittmann and Jäck, Genomics, transcriptomics, proteomics and metabolomics are representative “omics” disciplines of life science that deal with the entirety of genes, transcripts, proteins, and metabolites, respectively. Omics medicine is an emerging discipline of medical science that produces large amounts of omics data on genetics, genomics, epigenetics, transcriptomics, proteomics, and metabolomics. Here, I propose my personal view on the three-layer structure of omics medicine Figure . The firIndustry also consists of three layers. The first layer of industry includes agriculture, forestry, fishery, and mining; the second layer of industry includes manufacturing and construction; the third layer of industry includes financial business, commerce, service, information, and health care. The Industrial Revolution caused a shift from the first layer of industry to the second layer of industry during the 18th and 19th centuries, while the Internet Revolution and Lehman Shock promoted the prevalence of commodity-assembly manufacturing to reduce personnel expenses and material costs, causing another rotation from the second layer of industry to the third layer of industry in the 21st century.There are many analogies between the three-layer structure and development process of omics medicine and those of industry. Internet technology enabled the outsourcing of diagnostics and therapeutic optimization, which are known as telemedicine. Microarray and next-generation sequencing technologies concentrated the production of high-throughput data to world-class institutes or companies to reduce personnel and consumable expenses. Because the internet and high-throughput technologies are able to promote a leap from the first layer to the third layer of omics medicine, the global scientific community appears destined to move toward translational medicine.Clinical medicine, basic medicine and translational medicine are responsible for the establishment and maintenance of bio-banks, databases and a comprehensive knowledgebase, respectively Figure . All of