Moniliophthora perniciosa causes witches’ broom disease in cacao. As a hemibiotrophic fungus, it initially colonizes the living host tissues (biotrophic phase), and later grows over the dead plant (necrotrophic phase). Little is known about the mechanisms that promote these distinct fungal phases or mediate the transition between them.The tropical pathogen (Mp-aox) was identified in the M. perniciosa genome and its expression was analyzed througout the fungal life cycle. In addition, the effects of inhibitors of the cytochrome-dependent respiratory chain (CRC) and alternative oxidase (AOX) were evaluated on the in vitro development of M. perniciosa.An alternative oxidase gene Mp-aox transcripts were observed in the biotrophic hyphae, which accordingly showed elevated sensitivity to AOX inhibitors. More importantly, the inhibition of CRC prevented the transition from the biotrophic to the necrotrophic phase, and the combined use of a CRC and AOX inhibitor completely halted fungal growth.Larger numbers of M. perniciosa and regulates the transition to its necrotrophic stage. Strikingly, this model correlates well with the infection strategy of animal pathogens, particularly Trypanosoma brucei, which uses AOX as a strategy for pathogenicity.On the basis of these results, a novel mechanism is presented in which AOX plays a role in the biotrophic development of  Fungal plant pathogens have traditionally been classified on the basis of their feeding mechanism into biotrophic, necrotrophic and hemibiotrophic pathogens. Biotrophs are defined by a dependence on the host to complete their life cycle; they derive nutrients from living host cells by forming specialized infection structures, such as haustoria. By contrast, necrotrophs kill the plant and derive nutrients from the dead host cells. In some pathosystems, the pathogen initially keeps cells alive (biotrophic phase), but then kills them at later stages of infection (necrotrophic phase), which characterizes the hemibiotrophic lifestyle . DespiteMagnaporthe oryzae, which causes the rice blast disease, has emerged as a model for the study of phytopathogenic fungi because of its experimental tractability and economic importance  in M. oryzae  causes one of the most devastating diseases of cacao (Theobroma cacao), namely witches’ broom disease (WBD). The M. perniciosa–cacao interaction is classified as hemibiotrophic  phase. The first report of the successful maintenance of the M. perniciosa monokaryotic phase under laboratory conditions was in a study of a dual culture of basidiospores with potato callus  are not cultivable outside of their hosts , electron transfer through AOX does not involve phosphorylation, and the redox energy is released as heat instead of being used for ATP production , leading to the generation of deleterious oxidative stress . StudiesM. perniciosa aox gene was characterized, and a clear correlation between the hemibiotrophic lifestyle of this fungus and the functionality of the mitochondrial respiratory routes was observed. In addition, this work suggests that AOX may enable respiration to continue during the first stages of WBD, when the pathogen needs to survive in a hostile environment established by the host defense system (e.g. NO burst). Finally, we show that the inhibition of CRC maintained the fungus in its monokaryotic stage, which may be sustained by AOX-dependent respiration. These data indicate that mitochondrial respiratory pathways and cellular energetic status may play a role in the phase transition and development of phytopathogens.In this study, we present strong evidence for a biological role of AOX in the life cycle of a plant pathogen. The Moniliophthora perniciosa (Stahel) −1 malt extract, 5 g l−1 yeast extract and 20 g l−1 agar) induced the transition to the necrotrophic phase. Cultures were maintained at 28°C with agitation of 120 rpm when cultured in liquid media.The experiments were performed using the Theobroma cacao L. variety ‘comum’ (Catongo type) was used for infections. Plants were grown for c. 3 months in a glasshouse under controlled temperature (26°C) and humidity (> 80%) and a photoperiod of 12 h. Active apical meristems were inoculated with 30 μl of a basidiospore suspension (105 spores ml−1), followed by incubation in a humid chamber for 24 h . Mitochondria were energized with 10 mM glutamate/malate, followed by the addition of 1 mM ADP. AOX activity was measured in the presence of the CRC inhibitor antimycin-A (10 μM), as the oxygen consumption inhibited by 100 μM n-propyl gallate.AOX activity was evaluated by measuring the oxygen consumption profile of mitochondria isolated from necrotrophic hyphae, the mycelial stage from which a sufficient amount of organelles can be extracted. Mitochondria isolation was conducted according to the protocol described by M. perniciosa, rat (negative control) and Arabidopsis thaliana (positive control) were resuspended in 1 × Laemmli buffer , 23 mM Tris-HCl pH 6.8, 0.01% bromophenol blue) and heated at 95°C for 5 min. Mitochondrial proteins (12.5 μg) were separated by sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to a polyvinylidene difluoride (PVDF) membrane using standard protocols. The membrane was blocked with 5% nonfat milk in TBST  for 1 h and then incubated with a 1 : 100 dilution of the anti-AOX monoclonal antibody (AOA) (c oxidase complex (COX3) was used as endogenous control to demonstrate homogeneity in protein loading of the M. perniciosa mitochondrial samples. Three micrograms per milliliter of the anti-yeast COX3 antibody (Invitrogen) were used in the immunoblotting, according to the same procedures as described for the AOA.Purified mitochondria from Mp-aox gene expression were evaluated by quantitative PCR (qPCR). To this end, necrotrophic hyphae were grown in liquid cultures  at 28°C and with agitation of 120 rpm. Seven days later, cultures were independently treated with the different compounds for 4 h and then harvested for RNA extraction. For the analysis of Mp-aox gene expression along the M. perniciosa life cycle, the different fungal stages were obtained as described in the ‘Biological material’ section above. To avoid variations in gene expression associated with different growth conditions, biotrophic and necrotrophic hyphae were grown in the biotrophic-maintaining medium. After 7 d of growth, both mycelia were collected to perform the qPCR assays.The effects of azoxystrobin  and of the NO donor 1-hydroxy-2-oxo-3-(3-aminopropyl)-3-isopropyl-1-triazene  on actin and the aox gene were amplified with the gene-specific primer pairs ACTRTF (5′-CCCTTCTATCGTCGGTCGT-3′) and ACTRTR (5′-AGGATACCACGCTTGGATTG-3′), and AOXRTF (5′-GACGTGCCTTTCGGATAGAG-3′) and AOXRTR (5′-CTTGCCAGGAGGAATGGTT-3′), respectively. All qPCRs were performed in a 16-μl mixture containing 1/16 volume of the cDNA preparation, 1/2 volume of SYBR Green Master Mix  and 0.2 μM of each primer. Real-time quantifications were executed using the StepOnePlus system (Applied Biosystems) and the following cycles: 2 min at 50°C, 10 min at 95°C and 40 cycles of 15 s at 95°C, 30 s at 53°C and 60 s at 60°C. Dissociation curves were performed to verify the formation of primer dimers and nonspecific amplifications. Data analysis was completed using the mathematical method proposed by Isolation of total RNA from fungal cells was performed using the Plant RNeasy Minikit (Qiagen), and its concentration and quality were assessed with a NanoDrop ND-1000 spectrophotometer . RNA integrity was evaluated on 1% formaldehyde-agarose gel. Synthesis of cDNA was performed with the Superscript II Reverse Transcriptase kit (Invitrogen) using 1 μg of DNAse I-treated total RNA. The housekeeping gene β-Mp-aox gene during the in planta development of M. perniciosa in green brooms and dry brooms.RNA-seq libraries produced as part of the WBD Transcriptome Atlas  were inspected for the expression of the Apical meristems of healthy plantlets and infected cacao seedlings at the green broom stage were collected to evaluate NO generation using the fluorescent indicator 4,5-diaminofluorescein (DAF-2) (M. perniciosa in the biotrophic and necrotrophic phases were tested ex planta. Azoxystrobin (2.5 μM – Syngenta) was used as a CRC inhibitor  was also tested.The effects of CRC and AOX inhibitors on the growth of aox gene in M. perniciosa (Mp-aox – GenBank accession number JF826501) is a single-copy gene of 1616 nucleotides and comprises ten exons and nine introns. Mp-aox encodes a protein of 378 amino acids that has the conserved motifs described for other plant and fungal AOXs (M. perniciosa AOX (Mp-AOX) and those described in other fungi (data not shown).The gal AOXs . MoreoveM. perniciosa necrotrophic mycelium with the CRC inhibitor azoxystrobin induces Mp-aox transcription  . Moreover, these data suggest that the enzyme is coordinately regulated at gene expression, protein production and activity levels.Gene expression and western blot analyses showed that treatment of the cription  and incrcription , as obsehibitor) . Convershibitor) , showingex planta expression profile of the Mp-aox gene to identify specific conditions during the development of M. perniciosa in which Mp-aox is activated. As shown in aox transcripts was verified in the monokaryotic biotrophic-like mycelium when compared with other fungal stages . Finally, supporting these results, inspection of the RNA-seq database  demonstrated that the M. perniciosa aox gene is over-expressed in green brooms, when compared with dry brooms, showing a direct correlation between the ex planta and in planta analyses , suggesting that CRC inhibition, which results in a low energetic status and possibly high AOX respiration, may favor biotrophic development. Furthermore, in the presence of SHAM  to further confirm the effects of AOX inhibition on fungal development. As expected, the biotrophic mycelium showed a higher sensitivity to this inhibitor in comparison with the necrotrophic mycelium .As reported previously . AnalysiThe necrotrophic mycelium developed in the presence of both respiratory inhibitors – azoxystrobin and SHAM – at the concentrations tested  of T. brucei greatly depends on the activity of the alternative route, and treatment with AOX inhibitors (SHAM/ascofuranone) results in trypanocidal effects  in the biotrophic-like mycelium suggest that the enzyme may play a role during this specific fungal stage. Accordingly, inspection of the WBD Transcriptome Atlas  showed that this gene is up-regulated during the green broom stage of WBD  when compared with the dry broom stage (which is colonized by the necrotrophic hyphae) (The increased expression of the aox gene  and the  hyphae) . DespiteM. perniciosa with protection against the harmful effects of NO and its derivatives on the mitochondrial respiratory chain. In support of this hypothesis, we verified that Mp-aox gene expression is up-regulated by the NO donor NOC-5 ex planta  for intense pathogen growth during the necrotrophic phase. In this regard, AOX may be a switch regulating the development of these two life strategies in M. perniciosa (biotrophic and necrotrophic phases). Our study is the first report of the participation of AOX in the development of a plant pathogen, which sheds light on the physiological functions of this enzyme as well as on the mechanisms that control the phase transition process in phytopathogens, a central step during infection.Previous work by M. perniciosa  during which the fungus is in continuous contact with the living plant tissues  than those described for other hemibiotrophic phytopathogenic fungi (M. perniciosa and T. brucei. As in M. perniciosa, the protozoan T. brucei uses an AOX-dependent respiratory pathway during its infective stage inside the mammalian host. However, during its reproductive cycle inside the insect vector, the protozoan’s respiration is primarily dependent on the activity of the cytochrome pathway (T. brucei overcome host defense mechanisms that are mainly based on the induction of oxidative stress (Although e stress .M. perniciosa life cycle, we believe that this enzyme is a promising target for the development of new chemicals to disrupt M. perniciosa development and to effectively control WBD. Whether the enzyme also plays a role in the development of other phytopathogens, particularly in other hemibiotrophic fungi, should be the subject of future studies.In conclusion, this work presents a novel mechanism describing the development of phytopathogens that has a remarkable parallel to the mechanisms evolved in animal parasites. In addition, we believe that our work contributes to the understanding of the alternative respiratory pathway, which appears to be involved in the hemibiotrophic life cycle of a phytopathogenic fungus. Finally, based on the importance of AOX in the


The identity of the Human T lymphotropic Virus type 1 (HTLV-1) receptor remained an unsolved puzzle for two decades, until the recent demonstration that three molecules, Glucose Transporter 1, Neuropilin-1 and Heparan Sulfate Proteoglycans are involved in HTLV-1 binding and entry. Despite these advances, several questions remain unanswered, including the precise role of each of these molecules during virus entry. In light of the most recent data, we propose a model of the HTLV-1 receptor complex and discuss its potential impact on HTLV-1 infection. Since its identification in 1979, the Human T-Lymphotropic Virus type 1 (HTLV-1) has been the subject of extensive research. Despite abundant experimental data, several unsolved mysteries still surround HTLV-1. Amongst those, the identity of the HTLV-1 cellular receptor has eluded scientists for over two decades. Since 2003, data from several independent groups have shed light on the identity of the molecules that appear to be directly involved in the process of HTLV-1 entry. These include Glucose Transporter 1 (GLUT1)  and Hepaenv gene and antibodies directed against gp46/SU partially or totally abolished HTLV-1 infection in vitro [It is widely believed that HTLV-1 enters cells in a manner similar to that of other retroviruses including the well-studied HIV-1. HTLV-1 infection of target cells is believed to require the two virally encoded envelope glycoproteins (Env), the surface subunit (SU) gp46 and the transmembrane subunit (TM) gp21, generated from the cleavage of a polyprecursor (gp61) in the Golgi apparatus . As has in vitro ,13. Infein vitro . Altogetin vitro .5 are actually infectious [Experimental demonstration that specific molecules function as a virus receptor often is a challenge . This isfectious . Lastly,Virus tropism, the ability of a virus to replicate in particular cells, depends on interactions between viral components and cellular factors at each step of the viral cycle from the initial entry to the ultimate release and transmission of virions. Here, we will focus on the cellular factors that allow HTLV-1 entry, which is determined by the distribution of the receptor molecule(s).+ T cells [in vivo including as CD8+ T cells [In vivo, the only known targets of virus-induced transformation are CD4+ CD45RO+ memory T cells [Although the main targets of HTLV-1 are CD4 T cells , the vir T cells , monocyt T cells , macroph T cells , dendrit T cells ,24 and e T cells . In vivo T cells . Recentl T cells ), GITR+  T cells ,29. If t T cells ,31. This T cells ,33. This T cells . In cont T cells , which min vivo, molecules capable of supporting the initial steps in infection, binding and entry, appear to be expressed on nearly all established cell lines. As discussed below, this is consistent with the fact that molecules in the receptor complex are up-regulated on most transformed cells. Moreover, the HTLV-1 receptor complex is present on cell lines from nearly all known vertebrate species [In striking contrast to the limited number of cell-types in which HTLV-1 is detected  species : most av species . Further species . More re species . This st+ T cells but was rapidly upregulated upon activation. The receptor was also absent on naive T cells isolated from cord blood but could similarly be upregulated after stimulation by interleukin-7. Finally, it was reported that the SU-Fc could inhibit a mixed lymphocyte reaction [in vitro, demonstrating that molecules capable of both binding and fusion were expressed [per se was not required for receptor expression. This might constitute a strategy for the virus to increase its infectivity [+ T cells are known to abundantly secrete TGF-β.Although its ubiquitous nature considerably complicated its identification, several properties of the HTLV-1 receptor, in particular its expression pattern on primary cells, were characterized using indirect approaches. In 2003, two independent groups generated soluble SU proteins by generating recombinant SU-Ig-Fc fusion proteins containing either the full-length SU  or the Nreaction . This lareaction . The patxpressed . TGF-β-tectivity  as HTLV-Over the years, several molecules were proposed as candidate HTLV-1 receptors. It was proposed that the HTLV-1 receptor is encoded by chromosome 17q23.2-23.5, which was questioned in later studies ,43. Mostth extracellular loop (ECL6) of GLUT1 [It was not until 2003 that work from Sitbon and Battini's laboratory identified a candidate that matched all the prerequisites to be a HTLV-1 receptor ,52,53 alof GLUT1 .Further data from other laboratories confirmed that GLUT1 plays a role in HTLV-1 entry. TGF-β, which had previously been shown to upregulate molecules capable of binding HTLV SU, induces GLUT1 at the surface of T cells . OverexpHowever, several studies suggested that the identification of GLUT1 was not the end of the story. The astrocytoma/glioblastoma U87 cell line, which expresses very low levels of GLUT-1 due to a transdominant negative mutation in one of the alleles of GLUT1 (GLUT-1 D5), is easily infected by HTLV-pseudotyped viruses,56. MoreMolecules of the HSPG family are composed of a core protein associated with one or more sulphated polysaccharide side chains called heparan sulfate glycosaminoglycans. Because of their highly negative charge, HSPG bind through electrostatic interactions to a plethora of proteins including growth factors and their receptors, chemokines, cytokines and numerous proteins of the extracellular matrix or the plasma, thereby playing a major role in mammalian physiology . Pathoge+ T cells in Jones and Ruscetti's laboratory [+ T cells, HSPG augment both the binding of the SU-Fc and the infectious titer of pseudotyped viruses [The important role of HSPG in mediating attachment and entry has also been demonstrated for HTLV-1. Enzymatic removal of HSPG or inhibition of electrostatic interactions with dextran sulfate decreases the binding of full length soluble SU, Env-mediated syncytium formation and infection with pseudotyped viruses . These iboratory . Althougboratory , they ar viruses . The imp viruses ,6 and siOther recent studies suggest another role for HSPG during cell-cell transmission of the virus. It was discovered that HTLV-1 virions are stored outside the cell, within a protective microenvironment enriched for specific components of the extracellular matrix, including lectins and HSPGs . Followi+ T cells or CD8+ T cells, respectively, is governed by Env [+ T cells express a high level of HSPG and a lower level of GLUT1 whereas CD8+ T cells express a high level of GLUT1 but very few HSPG. In this model, the differences between HTLV-1 and HTLV-2 would be similar to what has been described in CD4-dependent and independent HIV-2 or SIV strains, which differ in their requirements for the binding receptor (CD4) but share similar fusion receptors [Interestingly, HSPG are important for infection with HTLV-1 but not HTLV-2 , showingd by Env . The difeceptors .165. NRP-1 was initially identified as a critically important factor in embryonic neuron guidance, and later shown to be a key player in the regulation of angiogenesis. In addition, Romeo and Hermine's laboratories were the first to demonstrate that NRP-1 is also involved in the regulation of the immune response [in vivo. NRP-1 is highly expressed on plasmacytoid DC (pDC), which can be infected by cell-free virus in vitro [in vivo [in vivo, NRP-1 is found in many tumor cells [In 2006, the Hermine and Pique laboratories showed that Neuropilin-1 (NRP-1) also displayed properties expected for a HTLV-1 receptor . This hyresponse . NRP-1 iresponse . NRP-1 iin vitro . Endothe[in vivo , also ex[in vivo . NRP-1 i[in vivo ,74. In cor cells  and hencor cells .Ghez et al. reported that NRP-1 was able to directly interact with the HTLV-1 and -2 SU . This inin vivo. This is also the case of GLUT1, whose expression is ubiquitous as well. In contrast, the distinct pattern of NRP-1 expression in vivo and in vitro may explain the disparity between the in vivo and in vitro tropisms. In primary cells, NRP-1 is mainly expressed on endothelial cells, activated CD4+ T cells and DC, all of which have been observed to be infected in vivo. However, NRP-1 overexpression upon cell transformation renders it nearly ubiquitous among established cell lines. Moreover, the nrp-1 gene is strongly conserved between mammalian species and has no homolog in insects. These findings strongly favor the notion that the in vivo HTLV-1 tropism is mainly dictated by NRP-1 expression.The identification of three new molecules involved in the process of HTLV-1 entry allows one to revisit the HTLV-1 tropism paradox mentioned above. While HSPG usage appears to distinguish HTLV-1 and HTLV-2, their ubiquitous expression cannot explain the limited tropism of HTLV-1 Thus, after more than 20 years, three different molecules have proven to be important for HTLV-1 entry. It seems clear that HSPG, as for nearly all other viruses, is involved in binding but not in fusion. The properties displayed by both GLUT1 and NRP-1, i.e. binding to the HTLV-1 SU and modulation of infection, are consistent with those expected for a receptor but, as mentioned above, many questions on their exact function remain unsolved. In particular, it is not clear whether GLUT1 and NRP-1 cooperate to promote HTLV-1 entry. We will review very recent data that have started to clarify the respective roles of HSPG, NRP-1 and GLUT1 in HTLV-1 infection.et al. determined that the minimal RBD is located within the first 183 amino terminal residues of the HTLV-1 SU [As was hypothesized previously, there is now definitive evidence that GLUT1 and NRP-1 bind to distinct regions of SU. Kim TLV-1 SU . MutatioTLV-1 SU , but hasTLV-1 SU . It is tTLV-1 SU . It is t165 (exon-7 peptide), decreased the DC-mediated transfer of HTLV-1 to CD4+ T cells by 50%. Moreover, blocking both HSPG and NRP-1 interactions decreased HTLV-1 infection by 80% [The HSPG-interacting SU region is located outside the GLUT1 and NRP-1 binding sites, and thus should not decrease its affinity to the other two molecules; this interpretation is consistent with the data showing that increasing cell surface levels of HSPG increases SU binding to receptor-expressing cells ,7. Blockn by 80% . Thus, Hn by 80% .et al. found that the binding of the HTLV-1 SU to NRP-1 involved two distinct types of interaction: an indirect interaction mediated by HSPG and a direct interaction mediated by the amino-acids 90-94 of the SU [165, [165. Interestingly, the aa 90-94 motif is conserved in the SU proteins of HTLV-2, HTLV-3 and related Simian T-Lymphotropic Viruses but not other retroviruses [et al. showed that mutating either aa 94 (R) or 114 (Y), which inhibits direct interaction with NRP-1 or GLUT1 respectively, is equally efficient in preventing interference due to overexpression of the H1-RBD in target cells [In 2009, Lambert SU [165, . This deoviruses . Strikinet cells . This re165, HSPG, and receptor . This binding to NRP-1 could trigger a conformational change within the SU, exposing the region necessary for GLUT1 binding including the critical Y114. Finally, the interaction of the SU with GLUT1 could trigger the fusion process necessary to HTLV-1 entry (step 3). In this model, NRP1, with the help of HSPG, is the primary binding receptor, while GLUT1 is the fusion receptor. This is consistent with the formation of NRP-1/Env/GLUT1 complexes and with the fact that GLUT1 over expression does not increase the level of HTLV-1 SU binding. This model applies to both primary CD4+ T cells and DC [In light of these data, it is possible to draw a model for HTLV-1 entry based on an analogy with HIV CD4/CXCR4 or CCR5 receptor complexes, and what is known about interactions of NRP-1 with its ligand VEGFAs and DC . Recent try step . In thisWhether this model is relevant for HTLV-1 entry in other types of cells remains to be investigated. A recent paper from Alkhatib's group showed that GLUT1, but not NRP-1, was involved in HTLV-1 entry into Hela cells . Moreovein vivo tropism of the virus better than the GLUT1-only hypothesis.Other evidence supporting a NRP-1/GLUT1- complex comes from reports that a small intracytoplasmic protein called GLUT1CBP or PSD-95/Dlg/ZO-1 can bind both NRP-1 and GLUT1 via its PDZ domain ,81. MoreGiven its role in the immune system, the discovery of NRP-1 as an entry molecule for HTLV-1 opens interesting perspectives on viral transmission and the physiology of HTLV-1 related diseases.+ T cells upregulates NRP-1 and HSPG within an hour followed by transmission of HTLV-1 to T cells and productive infection. This phenomenon appears to be dependent on both NRP-1 and HSPG expression on the target T cells [cis (from virus produced from infected DCs) but also in trans (prior to infection of the DC), as has been previously shown for DC-mediated HIV infection. This raises the question of whether DC (in particular pDC) could constitute a reservoir for the virus. Several laboratories have reported detecting HTLV-1 sequences in DC in vivo [in vivo.Plasmacytoid DC (pDC), and to a lesser extent myeloid dendritic cells (myDC), constitutively express high levels of NRP-1, which is involved in the regulation of DC-T cell interactions at the immune synapse . Shaking T cells . Viral t in vivo . It woulin vitro data demonstrated that cell-free HTLV-1 generates a pDC innate immune response by an induction of costimulatory molecules, production of massive levels of IFN-α, and rapid expression of the apoptotic ligand TRAIL. The role of NRP-1 in this process remains to be clarified. Furthermore, HTLV-1-stimulated pDC was shown to induce apoptosis of CD4+ T cells expressing DR5, transforming pDC into Interferon-producing Killer pDC (IKpDC) [Targeting DC and interfering with NRP-1 signalling might have several other consequences on viral infection. HTLV-1 infection results in a depletion of myDC as well as pDC and an impairment of their IFNα-producing capacity . Further (IKpDC) . Interac (IKpDC) . This ph165 is a potent competitor of HTLV-1 SU binding and HTLV-1 infection of primary T cells and DC in vitro [165 secretion by HTLV-1-infected cells [165 might not only have a negative effect. Sherer et al. showed in an elegant model of MLV infection that target cells projected filopodiae towards MLV infected cells [in vivo [165 [165 or Env itself might be envisaged with HTLV-1.Data clearly show that VEGFAin vitro ,72. VEGFed cells  might exed cells , VEGFA16ed cells . These e[in vivo  and coexivo [165 . Althoug165 production may participate in cell growth and angiogenesis [165 interaction may occur because the virus Env is no longer expressed in vivo.In ATLL it has been shown that VEGFAogenesis ,92. At tAnother major NRP-1 ligand, sema3a, could also play a role in the regulation of HTLV-1 infection. Polarization of the cytoskeleton is a critical determinant of transmission in a number of retroviruses including HIV-1 and HTLV-1 . Althoug+ T cells provides fascinating perspectives for future research. Further work is clearly needed to better define the precise role of the elements of this "ménage à trois" and its consequence on infected individuals. A better understanding of the effect of HTLV-1 infection on the immune system could lead to the development of alternative strategies in the treatment of HTLV-1-related diseases early in the course of the infection or even at a later stage, as in ATLL.The recent discovery that GLUT1, NRP-1 and HSPG are involved in HTLV-1 entry sheds new light on HTLV-1 infection and the physiopathology of HTLV-1-related diseases. The demonstration that HTLV-1 uses molecules involved in the immune response and can infect dendritic cells more efficiently than CD4The authors declare that they have no competing interests.OH and DG designed the manuscript. DG wrote the manuscript. YL, KSJ, CP revised and added new information to the manuscript. OH coordinated the work and finalized the manuscript with DG.


Human T-cell Leukemia Virus type 1 (HTLV-1) is the etiological agent of tropical spastic paraparesis/HTLV-associated myelopathy (HAM/TSP) that can be identified in around 0.25%–3.8% of the infected population. Disease progression can be monitored by the proviral load and may depend on genetic factors, however, it is not well understood why some HTLV-1 infected people develop the disease while others do not. The present study attempts to assess the molecular diversity of gp46 glycoprotein in HAM/TSP patients and Health Carrier (HC) individuals.Blood samples were collected from 10 individuals, and DNA was extracted from PBMCs to measure the HTLV-1 proviral load. The gp46 coding sequences were amplified PCR, cloned and sequenced. The molecular characterization was performed using bioinformatics tools.6 PBMCs). The gp46 molecular characterization of 146 clones (70 HC and 76 HAM/TSP) revealed an overall diversity, within HC and HAM/TSP clones, of 0.4% and 0.6%, respectively. Five frequent mutations were detected among groups (HAM/TSP and HC clone sequences). A single amino acid (aa) substitution (S35L) was exclusive for the HC group, and three gp46 substitutions  were exclusive for the HAM/TSP group. The remaining frequent mutation (V247I) was present in both groups (p = 0.0014). The in silico protein analysis revealed that the mutated alleles F14S and N42H represent more hydrophilic and flexible protein domains that are likely to be less antigenic. The Receptor Binding Domain is quite variable in the HAM/TSP group. Two other domains (aa 53–75 and 175–209) that contain multiple linear T-cell epitopes showed genetic diversity in both HAM/TSP and HC groups. Further analysis revealed 27 and 13 T-cell epitopes for class I HLA alleles and class II HLA alleles, when analyzing the entire gp46.The median HTLV-1 proviral load of HC (n = 5) and HAM/TSP (n = 5) patients was similar (average 316,227 copies/10The most common gp46 mutations were not associated clinical status because they were found in only one individual, except for the V247I mutation, that was found at viral clones from HAM/TSP ad HC individuals. Because of this, we cannot associate any of the gp46 found mutations with the clinical profile. HTLV-1 was initially isolated from a patient with cutaneous T-cell lymphoma in early 1980s, being the first discovered human retrovirus . HTLV-1 *08 (HLA class I) are associated with a significant reduction in both HTLV-1 proviral load and the risk of HAM/TSP, whereas HLA-B*5401 (HLA class I) and HLA-DRB1*0101 (HLA class II) predispose to HAM/TSP in the same population .All The authors declare they have no competing interests.ACAMM carried out the molecular genetic studies, the bioinformatics analyses and drafted the manuscript. FKB participated in the molecular genetic studies and the bioinformatics analyses. MFCA participated in the cloning. EB participated in the cloning and molecular studies. JPMC had performed the English review and helped to draft the manuscript. LFV contributed to the analysis of the results and helped to draft the manuscript. BGCF contributed with the blood samples and clinical data from HTLV-1 infected individuals. LCJA conceived of the study, participated in its design and coordination and helped to draft the manuscript. All authors read and approved the final manuscript.Brief description of previously published domains in the HTLV-1 gp46 protein and their exact aminoacid location.Click here for file


Jatropha curcas L. is a highly drought and salt tolerant plant species that is typically used as a traditional folk medicine and biofuel crop in many countries. Understanding the molecular mechanisms that underlie the response to various abiotic environmental stimuli, especially to drought and salt stresses, in J. curcas could be important to crop improvement efforts. In this study, we cloned and characterized the gene for a late embryogenesis abundant (LEA) protein from J. curcas that we designated JcLEA. Sequence analyses showed that the JcLEA protein belongs to group 5, a subgroup of the LEA protein family. In young seedlings, expression of JcLEA is significantly induced by abscisic acid (ABA), dehydration, and salt stress. Subcellular localization analysis shows that that JcLEA protein is distributed in both the nucleus and cytoplasm. Moreover, based on growth status and physiological indices, the overexpression of JcLEA in transgenic Arabidopsis plants conferred increased resistance to both drought and salt stresses compared to the WT. Our data suggests that the group 5 JcLEA protein contributes to drought and salt stress tolerance in plants. Thus, JcLEA is a potential candidate gene for plant genetic modification. Triticum aestivum) and cotton (Gossypium hirsutum), and were subsequently detected in many other species of higher plants Water and ion concentrations in soil are important abiotic elements for living organisms. Unlike animals, plants are sessile organisms that are exposed to seasonal and local environmental variations, and their survival and growth are strongly influenced by local stress factors. It has been estimated that 70% of crop yield losses are caused by abiotic stresses, with drought and high salinity being the most serious threats to crop production in many areas of the world LEA proteins are localized to various subcellular compartments, including the nucleus, mitochondria, chloroplast, endoplasmic reticulum, vacuole peroxisome, plasma membrane and cytoplasm LEA genes have been cloned and characterized from many plant species, including cotton LEA D-34 Craterostigma plantagineum pcC27-45 Medicago truncatula MtPM25 The LEA protein super families have a diversity of sequences and functions. Based on the nomenclature introduced by Battaglia, LEA proteins are categorized into seven distinctive groups, which include groups 1 (LEA-5), 2 (dehydrin), 3 (LEA-4), 4 (LEA-1), 5 , 6 (LEA-6), and 7 (ABA-WDS) Jatropha curcas L. is a multi-purpose shrub belonging to the plant family Euphorbiaceae. The species is widespread throughout arid and semi-arid tropical regions of the world, and has been utilized as a source of traditional folk medicines. J. curcas is so highly tolerant to dehydration that it can survive 30% of the field capacity (FC) without drought injury J. curcas has an efficient adaptive mechanism that enables it to tolerate severe drought situations by maintaining leaf water status and making effective osmotic adjustments J. curcas possesses a moderate tolerance to salinity, because the seedlings can tolerate up to 30 mM NaCl (in irrigation water) without negative effects on their growth parameters. Salt-treated plants (irrigated with NaCl levels of over 30 mM NaCl) showed a significant reduction in growth (by 5.82% for every 10 mM increment in NaCl concentration). This finding shows that J. curcas was more saline-tolerant than most of the typical Mediterranean crops J. curcas to environmental stresses, especially the functions of LEA proteins in protecting seedlings from the effects of dehydration and high salinity, are still poorly understood. In the present report, we document the molecular cloning of an LEA-like gene from J. curcas by RACE-PCR. Bioinformatics and expression analyses reveal that the J. curcas LEA protein strongly resembles group 5 LEAs from other plant species. The results of physiological assays under drought and salt treatments revealed that the JcLEA gene confers significantly enhanced tolerance to drought and salt in transgenic Arabidopsis plants. Based on these results, the potential application of the JcLEA gene in genetic engineering of crops is discussed.Jatropha curcas were obtained from the Jatropha curcas Germplasm Resources Preservation Center. The plants were grown in a natural environment on farmland in the Renhe . No specific permission was required for any of the locations or activities, and the field studies did not involve endangered or protected species. Seeds were germinated in soil in the greenhouse. Roots, leaves and stems were collected and stored at −70°C. Seeds of Arabidopsis thaliana accession Columbia (Col-0) were obtained from the Arabidopsis Biological Resource Center .Seeds of J. curcas plants were treated with 300 mM NaCl for 1 day, 100 µM ABA (Abscisic acid) for 3 hours and 30% PEG-4000 for 3 days, respectively. For root length determination, surface-sterilized Arabidopsis seeds were germinated on MS (Murashige and Skoog) medium (pH 5.8) containing 2% phytagel with 50 mM, 100 mM, and 150 mM NaCl or 10% and 15% PEG-4000. The seedlings were grown under a 16 h light/8 h dark photoperiod at a temperature of 22°C with a light intensity of 150 µmol m−2 s−1 in a plant growth cabinet. Plants were exposed to drought and salt stress for 14 days and then photographed, and root length was measured using ImageJ software.For gene expression analysis, 30-day-old Arabidopsis lines overexpressing JcLEA were used for physiological tests of tolerance to high salinity and dehydration. Seedlings growing in the greenhouse under a 16 h light/8 h dark cycle at 22°C for 4 weeks were watered with 50 mM, 100 mM, 150 mM, or 200 mM NaCl for 25 days and photographed and analyzed for survival rates at the 25th day of salt treatment. For drought treatment, water was withheld from 4-week-old plants for 14 days, after which they were rewatered for 10 days. The survival rate determinations and physiological measurements were carried out at the 14th day under the drought treatment and the 10th day after rewatering. Physiological measurements including the relative water content, electrolyte leakage, glucose content, sodium content, and potassium content were measured at the 14th day of the dehydration treatment, the 10th day after rewatering, and the 25th day of salt treatment. Whole plants were used for investigation of sodium content and potassium content, and leaves were used for the other physiological measurements.Two transgenic J. curcas was extracted as previously reported A. thaliana leaves using the Plant RNA Mini Kit . RNA quality and concentration were analyzed by agarose gel electrophoresis  and spectrophotometry . The RNA samples were stored at −70°C prior to use.Total RNA of JcLEA cDNA. This pair of primers corresponded was designed from highly conserved amino acid sequences of LEA proteins from Ricinus communis (XP_002520716), Glycine max (XP_003536910), Medicago truncatula (XP_003590628) and Gossypium hirsutum (P09444). Amplifications were performed at 94°C for 4 min, followed by 30 cycles of amplification . Following this, 5′- and 3′-rapid amplifications of cDNA ends were conducted using SMART technology (SMART™ RACE cDNA Amplification Kit) to produce a full-length JcLEA cDNA. The JcLEA cDNA was sequenced using the DYEnamic Direct dGTP Sequencing Kit  and a 373A DNA sequencing instrument. The sequences were then compared with known sequences in the NCBI database using blastp (Standard Protein-Protein BLAST) on NCBI (www.ncbi.nlm.nih.gov). The conserved domains were searched with RPS-BLAST (Search the Conserved Domain Database) in the NCBI database. The phylogenetic tree of LEA proteins sequences was constructed based on their conserved properties and homology. The representative protein sequences of seven previously reported groups of LEA proteins were obtained from the NCBI database (http://www.ncbi.nlm.nih.gov/). MEGA 5.0 was used as a computational phylogenetics tool to construct the maximum likelihood tree Primers LEAF and LEAR were used in a reverse transcription polymerase chain reaction (RT-PCR) to obtain a partial fragment of the JcLEA was amplified using the gene-specific primers JcLEA-B and JcLEA-X and Pfu DNA polymerase (Promega). The resulting fragment was digested with BamHI/XbaI and ligated into the corresponding restriction sites of the pCAMBIA1304 vector under control of the CaMV35S promoter. For production of the JcLEA-GFP fusion protein, the JcLEA coding region was amplified using primers LEA-GFPF and LEA-GFPR. The PCR product was digested with NcoI/SpeI and inserted into the pCAMBIA1302 vector. Expression of the JcLEA-GFP protein fusion was driven by the CaMV35S promoter. These constructs were sequenced to confirm the amplification fidelity and the exact insertion point of JcLEA in the pCAMBIA vectors.The open reading frame (ORF) fragment of Arabidopsis, Arabidopsis Col-0 was transformed using the Agrobacterium-mediated floral dip method. Agrobacterium tumefaciens strain LBA4404 carrying the plasmid 35S::JcLEA was cultured overnight to an OD600 of 1.2 to 1.6. The Agrobacterium cells were collected by centrifugation for 10 min at room temperature at 5000 rpm and then resuspended to an OD600 of approximately 0.8 in floral dip inoculation medium  ethanesulfonic acid, 50 g l−1 sucrose, 10 µg l−1 6- benzylaminopurine and 0.02% v/v silwet L-77, pH 5.7). The T1 plants were selected on hygromycin (30 mg l−1) and further confirmed by PCR using primers JcLEA-full-F and JcLEA-full-R. T2 lines showing 3∶1 segregation were carried forward to the T3 generation. PCR positive homozygous T3 and T4 lines were used for functional analyses of JcLEA.For stable transformation of Nicotiana tabacum cells, leaves of 4-week-old sterilized tobacco plants were infiltrated with A. tumefaciens GV3101 carrying the JcLEA-GFP fusion vector. The leaves were placed on MS solid medium in the dark at 25°C for two days before the imaging analysis.For transient gene expression in Fluorescence signals were imaged using a confocal laser scanning microscope  at a magnification of 20× and analyzed with Zen software. Before the detection of the fluorescence signals, the tobacco leaf pieces were stained with DAPI for 40 min to facilitate the observation of leaf cell nuclei.N. tabacum leaf disks using a Nuclear and Cytoplasmic Protein Extraction Kit . A 50 ng sample of the extracted protein was boiled in SDS gel loading buffer and electrophoresed on a 15% polyacrylamide gel for three hours. Proteins were then electroblotted onto PVDF membrane using the Mini Trans-Blot® Electrophoretic Transfer Cell System (Bio-Rad). The membranes were then blocked in Tris-HCl-buffered saline : 10 mM Tris-HCl, 150 mM NaCl, pH 7.5) containing 1% bovine serum albumin (BSA) for ≥2 h. The anti-GFP antibody  was incubated with the membranes at a 1∶200 dilution in TBS containing 3% BSA overnight at 4°C. After washing, a 1∶5000 dilution of anti-rabbit IgG HRP secondary antibody (#E1012 Santa Cruz Biotechnology) was incubated for 1 hour at room temperature in TBST containing 3% BSA. The filter was then washed and incubated with a WestDura Femto ECL kit , and bands were visualized using a ChemiScope 3400 Mini Imaging System .Plant nuclear and cytosolic proteins were extracted from actin gene from J. curcas was used as the internal control.Quantitative real-time PCR (qPCR) was used to analyze gene expression levels as described previously JcLEA gene was analyzed in four transformed lines of Arabidopsis by semi-quantitative RT-PCR using the primers JcLEA-full-F and JcLEA-full-R with the following PCR protocol: 94°C for 5 min, followed by 28 cycles of 94°C for 30 s, and 55°C for 30 s. The tubulin gene from Arabidopsis was used as the internal control.The relative expression of the t-test.Ion leakage was determined by the method of Zhou et al. t-test.Leaves from four plants of each Arabidopsis line were collected for each treatment and the fresh weights (FW) were determined. The samples were then incubated in 8 ml deionized water at room temperature for 12 h at room temperature. The turgid weights (TW) of the leaf samples were measured. All samples were oven-dried at 65°C for 30 h, and the dry weights were measured. The relative water content (%) was calculated as = 100%×(FW-DW)/(TW-DW) t-test.Glucose content was assayed with a Glucose (HK) Assay kit . Leaves from four different plants of each Arabidopsis line for each treatment were collected and incubated in 1.5 ml distilled deionized water and the absorbance of NADH at 340 nm versus distilled deionized water was measured using a BioPhotometer Plus  3:HClO4) and the mixed solutions were heated on the electro-thermal board to clarification. The potassium or sodium standard concentration liquid was diluted and added into flame photometer with the samples and the blank control. Potassium production was determined at 766 nm and sodium at 589 nm using a flame photometer . The potassium and sodium contents of the samples were calculated against standard curves for potassium and sodium.A minimum of 30 plants from each transgenic line were treated with 50 mM, 100 mM, and 150 mM NaCl. Whole plants were washed three times with distilled deionized water to remove any possible surface ions before drying, and 0.5 g to 1 g tissues samples were then used for measurements. Samples were digested in 30 ml mixed acid digestion solution . The full-length JcLEA cDNA comprised a 765 bp open reading frame (ORF), a 58 bp 5′-untranslated region (UTR), and a 247 bp 3′-UTR with a poly (A) tail (www.expasy.org). The predicted JcLEA protein contained 33.46% hydrophobic residues, including alanine, isoleucine, leucine, phenylalanine, and valine residues, and no cysteine, histidine or tryptophan residues.(A) tail . The ORFhttp://www.ncbi.nlm.nih.gov/) showed that there was a relatively high similarity between the predicted JcLEA protein and other plant LEA proteins. Notably, the LEAs identified in the blast analysis all belonged to the group 5 subfamily, and included the following nine plant proteins: RcLEA-D34 from R. communis (XP_002520716), AhLEA-5 from Arachis hypogaea (ADQ91843), MtLEA-D34 from M. truncatula (XP_003590628), CsLEA-D34 from Cucumis sativus (XP_004146999), GmLEA-D34 from Glycine max (XP_003536910), VvLEA-D34 from Vitis vinifera (XP_003632304), LEA34_GOSHI from G. hirsutum (p09444), AtLEA-5 from A. thaliana (BAB01464) and DcECP31 from Daucus carota (BAD86645). A number of gaps and insertions were needed to optimize the alignment. The amino acid identities between JcLEA and RcLEA-D34, AhLEA-5, MtLEA-D34, CsLEA-D34, GmLEA-D34, VvLEA-D34, LEA34_GOSHI, AtLEA-5, and DcECP31 were found to be 76%, 64%, 64%, 64%, 63%, 62%, 61%, 57%, and 57%, respectively. Moreover, three SMP-like motifs that are generally present in LEA proteins were also detected  plants. Homozygous seeds of lines 25–29 and lines 25–33 were collected for further investigation. One set was germinated on soil for physiological measurements, and another set was germinated on MS medium for root length observation under conditions of drought and salt stress.To study the contributions of e RT-PCR . The traArabidopsis plants, especially for root length, which is sensitive to dehydration  the fact that the gene is induced by exposure to drought and salt, (2) the subcellular localization of JcLEA and (3) the improved tolerance to dehydration and high salt levels observed in transgenic Arabidopsis plants.In summary, JcLEA is a novel LEA protein belonging to group 5. The constitutive expression of


Human T-lymphotropic virus type 1 (HTLV-1), a human retrovirus, is the causative agent of a progressive neurological disease termed HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). HAM/TSP is a chronic inflammatory disease of the central nervous system and is characterized by unremitting myelopathic symptoms such as spastic paraparesis, lower limb sensory disturbance, and bladder/bowel dysfunction. Approximately 0.25–3.8% of HTLV-1-infected individuals develop HAM/TSP, which is more common in women than in men. Since the discovery of HAM/TSP, significant advances have been made with respect to elucidating the virological, molecular, and immunopathological mechanisms underlying this disease. These findings suggest that spinal cord invasion by HTLV-1-infected T cells triggers a strong virus-specific immune response and increases proinflammatory cytokine and chemokine production, leading to chronic lymphocytic inflammation and tissue damage in spinal cord lesions. However, little progress has been made in the development of an optimal treatment for HAM/TSP, more specifically in the identification of biomarkers for predicting disease progression and of molecular targets for novel therapeutic strategies targeting the underlying pathological mechanisms. This review summarizes current clinical and pathophysiological knowledge on HAM/TSP and discusses future focus areas for research on this disease. Human T-lymphotropic virus type 1 (HTLV-1), the first human retrovirus discovered  ; Table 2Detection of anti-HTLV-1 antibodies in cerebrospinal fluid (CSF) is necessary for the diagnosis of HAM/TSP, based on the WHO diagnostic guidelines. CSF examination revealed mild lymphocyte pleocytosis in approximately one-third of cases as well as mildly elevated protein concentration and increased concentrations of inflammatory markers such as neopterin  was abnormal in 3/21 (14%) patients with HAM/TSP in a small series where spinal cord atrophy was reported mainly in the thoracic region , neuromyelitis optica (NMO), spinal cord compression , transverse myelitis, collagen vascular disease, Sjögren syndrome, hereditary spastic paraparesis, primary lateral sclerosis, subacute combined degeneration secondary to vitamin B12 and folate deficiency, human immunodeficiency virus-associated vacuolar myelopathy, neurosyphilis, and Lyme disease, among others. Differentiating, rapidly progressing HAM/TSP from NMO is important. NMO shows more rapid progression than HAM/TSP, and HAM/TSP usually does not present with optic neuritis. Importantly, from our clinical experience, HAM/TSP patients are negative for a specific diagnostic antibody for NMO termed NMO-IgG or anti-aquaporin-4 antibodies (data not published). Furthermore, differentiating HAM/TSP from primary progressive MS is occasionally a diagnostic challenge, since the two conditions are clinically indistinguishable and the mere presence of positive HTLV-1 serology does not necessarily lead to neurological disease. This diagnostic difficulty is compounded by the fact that sometimes, white matter abnormalities are found on brain MRI of HAM/TSP patients  cells under healthy conditions, are the predominant viral reservoir of HTLV-1 in both ATLL and HAM/TSP  cells in HAM/TSP patients are significantly lower than those observed in healthy controls . The disease usually progresses slowly without remission. However, there is a subgroup of patients with rapid progression who are unable to walk within 2 years, and another subgroup of patients with very mild progression , which have immunomodulatory and antiviral properties (Borden et al., in vitro, was conducted in 16 patients, and no significant changes were observed in the clinical symptoms and HTLV-1 proviral load (Taylor et al., in vitro, was conducted in six patients, and no significant change in HTLV-1 proviral load was observed (Macchi et al., in vivo has been minimal. These results support the hypothesis that HTLV-1 proviral load in HTLV-1-infected patients is mainly maintained through cell division of infected cells and not by viral replication and new infection (Wattel et al., Thus, corticosteroids and IFN-α may have therapeutic efficacy for HAM/TSP to some extent; however, the effect may not be sufficient for avoiding long-term disability. Moreover, in some cases, it might be difficult to continue therapy because of the side effects of these drugs and their insufficient benefit. Therefore, it is essential that revolutionary drugs that can lead to a paradigm shift in the therapeutic strategies for HAM/TSP be developed. Considering the pathogenesis of HAM/TSP, therapies to eliminate HTLV-1-infected cells from the peripheral blood and CNS should be developed. However, antiviral therapy has not been successful in the clinical trial for HAM/TSP. A randomized, double-blind, placebo-controlled, 6-month study of zidovudine and lamivudine combination therapy, which demonstrated activity against HTLV-1 reverse transcriptase Advances in study of the epidemiology and pathogenesis of HAM/TSP have led to the identification of several biomarkers and therapeutic targets. However, these findings have not yet translated into an optimal therapeutic strategy for this hitherto intractable neurological disease. Well-designed clinical trials in HAM/TSP will provide opportunities for further quantification of biomarkers and refinement of therapeutic drugs. The development of an effective therapy to improve long-term prognosis in HAM/TSP is of paramount importance, and clinical trials for the validation of HAM/TSP relevant biomarkers and new therapeutic targets will be key challenges in this therapy.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Diabloceratops) and China (Sinoceratops), plus otherwise fragmentary remains from the southern Western Interior of North America. Here, we describe a remarkable new taxon, Nasutoceratops titusi n. gen. et sp., from the late Campanian Kaiparowits Formation of Utah, represented by multiple specimens, including a nearly complete skull and partial postcranial skeleton. Autapomorphies include an enlarged narial region, pneumatic nasal ornamentation, abbreviated snout and elongate, rostrolaterally directed supraorbital horncores. The subrectangular parietosquamosal frill is relatively unadorned and broadest in the mid-region. A phylogenetic analysis indicates that Nasutoceratops is the sister taxon to Avaceratops, and that a previously unknown subclade of centrosaurines branched off early in the group's history and persisted for several million years during the late Campanian. As the first well-represented southern centrosaurine comparable in age to the bulk of northern forms, Nasutoceratops provides strong support for the provincialism hypothesis, which posits that Laramidia—the western landmass formed by inundation of the central region of North America by the Western Interior Seaway—hosted at least two coeval dinosaur communities for over a million years of late Campanian time.The fossil record of centrosaurine ceratopsids is largely restricted to the northern region of western North America . Exceptions consist of single taxa from Utah ( Laramidia witnessed the greatest radiation of Mesozoic dinosaurs documented to date , with Ce of Utah . Additio of Utah , the MenGryposaurus monumentensis [Hagryphus giganteus [Teratophoneus curriei [Talos sampsoni [Utahceratops gettyi and Kosmoceratops richardsoni [Since 2000, a collaborative, multi-institutional team working in southern Utah's Grand Staircase-Escalante National Monument has unearthed a previously unknown dinosaur assemblage from the late Campanian Kaiparowits Formation . A totalentensis , the oviiganteus , the tyr curriei , the trosampsoni  and a pahardsoni ,12. Here2.sensu Padian and May 1993Dinosauria Owen, 1842 sensu Sereno 1998 Ornithischia Seeley, 1887 sensu Dodson, 1997 Ceratopsia Marsh, 1890 sensu Sereno 1998 Ceratopsidae Marsh, 1888 sensu [ Centrosaurinae Lambe, 1915 sensu Nasutoceratops titusi n. gen. et sp.  urn:lsid:zoobank.org:act:F9997290-2618-4C95-9D46-7EED00C99916(a)nasutus, meaning ‘large-nosed’ and the Latinized Greek ceratops, meaning ‘horned-face’; titusi, honouring Alan Titus, palaeontologist at Grand Staircase-Escalante National Monument, for his exemplary efforts assisting palaeontological fieldwork in the Monument.From the Latin (b)Natural History Museum of Utah (UMNH) VP 16800, holotype consisting of a mostly complete, articulated 1.8 m long skull plus postcranial elements: syncervical, three fragmentary dorsal vertebrae, associated left forelimb and fragmentary right forelimb. Referred materials consist of UMNH VP 19466—a disarticulated adult skull including a partial premaxilla, maxilla and nasal—and UMNH VP 19469, an isolated squamosal.(c)Nasutoceratops occurs within the middle unit (approx. 250–320 m) of the Upper Campanian Kaiparowits Formation, dated to the late Campanian, approximately between 75.51 and 75.97 Ma . Detail(d)Nasutoceratops can also be distinguished by a unique suite of synapomorphies, including: low, transversely narrow, rostrocadually elongate nasal horncore; pronounced dorsolateral ridge on squamosal; subcircular parietosquamosal frill widest near mid-region; simple, crescentic episquamosals and epiparietals; and the presence of a midline epiparietal.Centrosaurine ceratopsid bearing the following autapomorphies: hypertrophied narial region ; caudal portion of fused nasals occupied by internal pneumatic cavity; uniquely enlarged premaxillary contact of maxilla; double faceted, medially directed flange on maxilla and rostrolaterally directed, rostrally curved and apically twisted supraorbital horncores; 3.Nasutoceratops below is limited to diagnostic aspects of skull anatomy.The lower level taxonomy of ceratopsid dinosaurs has been based almost exclusively on craniofacial characters. Thus, the abbreviated description of (a)Nasutoceratops. Overall, the preorbital facial skeleton is relatively short rostrocaudally, comprising less of the total skull length than in any other ceratopsid . Yet, the narial region, dominated by the premaxilla and ectonaris, is hypertrophied, comprising approximately 75% of preorbital skull length. The front of the snout is also expanded dorsally, resulting in an ‘inflated’ appearance compared with other centrosaurine taxa. Associated with expansion of the narial region is an enlarged premaxillary septum, also more extensive than in other centrosaurines.The bony anatomy of the narial region of centrosaurines, although derived relative to other dinosaurs, tends to be conservative within the clade. Yet, this region is unique in several respects in Nasutoceratops specimen UMNH VP 19466 includes a complete maxilla bearing 29 alveoli. The teeth are typical of centrosaurines. The maxillary tooth row of Nasutoceratops is displaced ventrally, much as in Albertaceratops, Avaceratops, Diabloceratops and many non-ceratopsid neoceratopsians. The premaxillary contact surface on the maxilla is exceptionally broad transversely, forming a deep concavity ventrally and becoming shallower dorsally. A double faceted, medially directed flange of the maxilla slots into premaxilla, forming a greater contribution to the hard palate than in related forms.The ascending ramus of the maxilla is rostrocaudally abbreviated and the maxillary body is dorsoventrally expanded, both features presumably related to the premaxillary expansion. The conformation of this morphology is reflected in the steeply inclined ascending ramus; centrosaurines typically possess a more caudally projected, rostrocaudally thickened ascending ramus. The holotype maxilla preserves 22 maxillary tooth positions, and the referred Nasutoceratops is somewhat foreshortened relative to the condition in more derived centrosaurines. The nasal horncore is relatively low, rostrocaudally elongate and bladelike (transversely narrow), situated dorsal to the endonaris. The bulk of derived centrosaurines exhibit more elaborate forms of nasal ornamentation, including large horncores of varying orientation  and pachyostotic bosses (Achelousaurus and Pachyrhinosaurus). The relatively diminutive horncore of Nasutoceratops, by contrast, more closely resembles that of the basal centrosaurine Albertaceratops [Nasutoceratops exhibit well-developed, caudally positioned internal cavities within the body of the nasal caudal to the horncore. These cavities are here interpreted to represent pneumatic excavations from the paranasal region. Pneumatic nasals are unknown in any other ceratopsid, and this feature is here regarded to be autapomorphic for the new Utah taxon.Like the maxilla, the nasal of ceratops . Both of(b)Nasutoceratops are highly distinctive, being exceptionally elongate, rostrodorsally oriented and twisted distally. Within Centrosaurinae, supraorbital horncores tend to be relatively short, the only exceptions being the basal taxa Diabloceratops, Avaceratops and Albertaceratops. Given that the sister taxon to Ceratopsidae, Zuniceratops, also exhibits elongate supraorbital horncores [Nasutoceratops are curved throughout their lengths, transitioning in orientation from rostrolateral to rostromedial at the tips, with the distal one-third exhibiting pronounced lateral torsion. This torsion combines dorsal curvature and twisting of the distal horncore, marked by longitudinal grooves along the external surface. In the holotype, UMNH VP 16800, the supraorbital horncores are relatively and absolutely the longest of any centrosaurine, spanning approximately 40% of total skull length and extending rostrally almost to the tip of the snout.The supraorbital horncores of orncores , this feNasutoceratops closely resembles that of the centrosaurines Albertaceratops and Diabloceratops [Kosmoceratops, Utahceratops and Pentaceratops [Caudal to the nasal horncore and immediately rostral to the orbits, the dorsum of the skull inclines dorsally to form a pronounced ‘forehead’ and vaulted skull roof composed largely of nasals, prefrontals, palpebrals and frontals. Whereas the majority of centrosaurines, and ceratopsids generally, possess a relatively flat skull roof between the facial skeleton and the cranium, the steeply inclined preorbital region of ceratops ,18, as wceratops ,12.Nasutoceratops holotype does include a well-preserved epijugal ossification. Whereas large epijugals are typical of chasmosaurine ceratopsids, centrosaurines tend to possess relatively small accessory ossifications on the distal jugal. By contrast, the epijugal of Nasutoceratops more closely resembles that of the basal centrosaurine Diabloceratops, in which this element is also strongly developed. The UMNH VP 16800 epijugal—relatively and absolutely the largest example known among centrosaurines—is approximately trihedral, with a flattened rostral surface. Given the distribution of large epijugals among chasmosaurines, some non-ceratopsid neoceratopsians (e.g. Protoceratops), and basal centrosaurines, this feature is probably symplesiomorphic for Ceratopsidae.Only a small dorsal portion of the jugal is preserved on UMNH VP 16800. However, the (c)Nasutoceratops resembles that of most centrosaurines , bearing a transversely convex and rostrocaudally concave dorsal surface. A large, oval parietal fenestra is present on either side, with the long axis oriented rostrocaudally. The Nasutoceratops frill is subrectangular as viewed dorsally, with the broadest point occurring in the mid-region. Total frill length, estimated at 610 mm in UMNH VP 16800, is approximately equal to basal skull length.In overall conformation, the parietosquamosal frill of Nasutoceratops closely resembles that of other centrosaurines in being relatively short rostrocaudally with a stepped caudomedial margin [Nasutoceratops differs in possessing a pronounced ridge on the dorsolateral surface, which, in UMNH VP 19469, can be seen to extend most of the element's length. Other centrosaurines (e.g. Avaceratops and Albertaceratops) possess raised bumps in this area, but a fully formed, elongate ridge in Nasutoceratops extends approximately three times the length of the laterotemporal fenestra. This distinctive feature is otherwise known only on a centrosaurine squamosal (NMMNH P34906) from the Fort Crittenden Formation of Arizona [The squamosal is poorly preserved in UMNH VP 16800, but a referred, mostly complete specimen, UMNH VP 19469, has been recovered from the same stratigraphic interval of the Kaiparowits Formation. The latter specimen demonstrates that the squamosal of l margin . However Arizona .Nasutoceratops and Avaceratops is distinctive in lacking both a median embayment along the caudal margin and well-developed marginal ornamentations. In most centrosaurines, the parietal margin exhibits a unique suite of ‘spikes’ (e.g. Styracosaurus and Einiosaurus), ‘hooks’ (Centrosaurus and Coronosaurus) and/or ‘horns’ (e.g. Pachyrhinosaurus). These structures, formed largely by accessory ossifications , occur even in basal members of the clade . Althouceratops  and the Nasutoceratops are relatively thin (4–19 mm), even along the outer margins. Surface bone texture on this element appears to exhibit a mosaic of striated and mottled types, associated with subadult and adult status, respectively [Nasutoceratops reflects an ontogenetic stage rather than the mature condition, other aspects of the specimen  are indicative of adult status. Moreover, given that the epiparietals show no indication even of incipient hypertrophy, the frill morphology is postulated here to approximate the mature condition. Finally, the size of both UMNH VP 16800 and UMNH VP 19466 are consistent with the absolutely largest Centrosaurus skulls and the presence of adult bone texture [The parietal transverse bar and lateral rami of ectively . Althoug texture ,23 on th4.Diabloceratops, from the early Campanian Wahweap Formation of Grand Staircase-Escalante National Monument, was the first named southern Laramidian exception [Nasutoceratops, from the overlying Kaiparowits Formation, represents the second example, as well as the first southern centrosaurine from the late Campanian.Until recently, the lack of centrosaurine remains discovered in the American southwest prompted some investigators to postulate their existence only in the northern region of the palaeolandmass Laramidia . Diablocxception . NasutocNasutoceratops, we added the scorings for this taxon into the matrix used for two recently published centrosaurines [Nasutoceratops can be confidently placed within Centrosaurinae on the basis of a suite of synapomorphies, including: premaxilla with pronounced ventral angle; subcircular narial region; presence of narial spine composed of nasal and premaxilla and rostrocaudally abbreviated squamosal with stepped caudomedian margin. However, despite its late Campanian age, approximately coeval with the highly derived northern taxa Styracosaurus and Centrosaurus, Nasutoceratops retains several primitive features  otherwise absent in all other late Campanian centrosaurines. Phylogenetic analysis  originated on Laramidia 90–80 Myr ago. Early forms such as Diabloceratops possessed diminutive nasal horncores, relatively elongate supraorbital horncores and frills adorned with a single pair of elaborate marginal ornamentations. The present study reveals a basal split that resulted in two clades, both of which persisted into the late Campanian.In order to assess the phylogenetic position of saurines ,25; see  Montana ,28. TogeDiabloceratops and Albertaceratops, retained diminutive nasal horncores and elongate supraorbital horncores, beginning approximately 77.5 Ma descendant taxa possessed elongate nasal horncores, abbreviated supraorbital horncores, and typically more elaborate frills. Spinops sternbergii is the oldest known member of the replacement assemblage, with subsequent taxa including Coronosaurus, Centrosaurus, Styracosaurus and Rubeosaurus. The most derived members of this clade are characterized by the possession of relatively low, thickened bosses in place of the skull roof horncores, as well as similarly ornate frills (i.e. Achelousaurus and Pachyrhinosaurus).One of these clades, currently known only from northern Laramidia, evolved more elaborate frills with multiple hypertrophied epiparietals per side. Whereas middle Campanian representatives of this clade, such as Avaceratops is the earliest known member of this second centrosaurine clade. Nasutoceratops, from the late Campanian of Utah, post-dating Avaceratops by approximately 2 Ma, is the latest-occurring form (The second centrosaurine clade, identified here for the first time, to our knowledge, appears to have followed a distinctly different evolutionary trajectory, retaining relatively short nasal horncores and elongate supraorbital horncores, but simplifying the frill. This pattern of evolutionary modification parallels that within chasmosaurines, which also appear to have de-emphasized frill ornamentation in favour of enlarged supraorbital horncores. The early Campanian ing form . TogetheAvaceratops–Nasutoceratops clade secondarily reduced frill ornamentations and elaborated the supraorbital horns, the Spinops–Pachyrhinosaurus clade secondarily reduced supraorbital horns while hypertrophying both the nasal horn and frill ornamentation, effectively distributing bony apomorphic signalling structures across the entire skull roof.Several hypotheses have been put forth regarding the function of ceratopsid horn and frill structures, but the consensus view of the past several decades is that they functioned in intraspecific signalling, with horns also used in combat with conspecifics ,30. ReceNasutoceratops provides additional support for the dinosaur provincialism hypothesis—the idea that distinct, coeval, latitudinally arrayed communities of theropods and ornithischians existed on Laramidia for more than 1 Myr of late Campanian time [Avaceratops is the sister of Nasutoceratops, the former occurs in sediments that precede the latter by several million years. By the time of Nasutoceratops, northern centrosaurines all belonged to a distinct clade with more elaborate frill ornamentations. Thus, in addition to representing a previously unknown taxon not found in northern Laramidia, the discovery of Nasutoceratops suggests that a distinct, persistent, previously unrecognized clade of centrosaurines inhabited southern Laramidia during this interval.ian time ,12,24,34Nasutoceratops represents the first example of a previously unknown Campanian radiation of southern Laramidian centrosaurines bearing elongate supraorbital horns and simple frills. First, robust evidence now exists of distinct, highly diverse northern and southern vertebrate communities on Laramidia [a priori reason to expect a substantially different pattern during the late Campanian, particularly given the many taxa unearthed in Alberta and Montana for this interval. In short, Nasutoceratops adds a critical element to a rapidly emerging evolutionary picture, offering the first glimpse into centrosaurine diversity on southern Laramidia during the Late Campanian.Considered in unison, four lines of evidences suggest that aramidia ,12,34. Saramidia ,26, becoaramidia , and we


HTLV-1 propagates in the host by two parallel mechanisms: infectious spread, i.e. direct transfer of virions from cell to cell via the virological synapse; and mitotic spread, i.e. proliferation of HTLV-1-infected T cell clones. During chronic infection, mitotic spread predominates, generating very large clones in some individuals. Yet the reasons for the remarkable variation within and between hosts in the abundance of HTLV-1-infected clones remain unknown.We aim to identify and quantify the selection forces that determine the size of HTLV-1-infected T cell clones in vivo and hence the risk of the associated inflammatory and malignant diseases. We shall summarize evidence from host and viral genetics, cellular immunology and high-throughput proviral integration site analysis that:1. The ‘quality’ or efficiency of a person’s genetically-determined cytotoxic T lymphocyte (CTL) response to the HBZ protein is a major determinant of that person’s proviral load and the risk of inflammatory diseases.2. The chief determinant of CTL quality is the host genotype in HLA Class 1 and Killer cell Immunoglobulin-like Receptor (KIR) loci.3. The CTL response to the Tax protein is immunodominant but does not protect against HAM/TSP.4. The site and orientation of the HTLV-1 provirus integrated in the T cell genome determine the size of each T cell clone in the host.5. Negative selection on the virus dominates during chronic infection: we postulate that this selection is exerted by CTLs.


Adult T cell Leukemia, an aggressive T-cell malignancy linked to HTLV-1 infection, is resistant to chemotherapy. Recently, promising results were obtained with the combination of arsenic trioxide, interferon alpha and zidovudine. However, the cellular and molecular mechanisms of their anti-leukemia activity remain to be investigated. To that aim, we have relied on HIS (Human Immune System) Rag2-/-γc-/- mice. We have indeed observed that when infected with HTLV-1, these mice displayed, five months later, an elevated number of human Tax-expressing T cells in the thymus, the spleen and the mesenteric lymph nodes. Some of them also developed T lymphoproliferative diseases.To determine the effects of these three drugs on the apparition of these pathological features, Rag2-/-γc-/- mice were infected with HTLV-1 and, 16 weeks after infection, daily treated with the drugs for one week and sacrificed. Untreated HIS mice infected with HTLV-1 were used as control. Treatment resulted in a significant decrease in the spleen weight as compared to untreated controls. Interestingly, we also noted a decrease of the proviral load in thymocytes as well as a drop in the number of mature activated T cells in the spleen and in lymph nodes. These data clearly indicate that the HIS mouse model provides us with the opportunity not only for the pre-clinical evaluation of therapeutic approaches against the leukemogenic process associated with HTLV-1 infection, but also for unraveling the corresponding mechanisms.


Following myocardial infarction, wound healing takes place in the infarct area where the non-viable cardiac tissue is replaced by a scar. Inadequate wound healing or insufficient maintenance of the extracellular matrix in the scar can lead to excessive dilatation of the ventricles, one of the hallmarks of congestive heart failure. Therefore, it is important to better understand the wound-healing process in the heart and to develop new therapeutic agents that target the infarct area in order to maintain an adequate cardiac function. One of these potential novel therapeutic targets is Wnt signaling. Wnt signaling plays an important role in embryonic myocardial development but in the adult heart the pathway is thought to be silent. However, there is increasing evidence that components of the Wnt pathway are re-expressed during cardiac repair, implying a regulatory role. Recently, several studies have been published where the effect of interventions in Wnt signaling on infarct healing has been studied. In this review, we will summarize the results of these studies and discuss the effects of these interventions on the different cell types that are involved in the wound healing process. Cardiovascular diseases (CVDs) are the leading causes of mortality worldwide. More people die from CVDs than from any other disease. According to WHO, 17.3 million people died from CVDs in 2008, representing 30% of all global deaths. Of these deaths, 7.3 million were due to coronary heart disease [Myocardial infarction (MI) is the result of an occlusion of a coronary artery, caused by a thrombus or an atheromatous ruptured plaque, which deprives the myocardium from sufficient blood flow. This leads to ischemia and eventually death of cardiomyocytes . The locWound healing is a complex process of consecutive cascades involving several cell types, and finalizes with scar formation. The first phase begins when necrotic cardiomyocytes trigger an inflammatory response by activation of the complement cascade . In humaMany cell types contribute to the process of infarct healing, having either a beneficial or detrimental role. Adverse remodeling of the infarct scar will ultimately result in congestive heart failure (CHF). Current therapies are only focused on the delay of the progression of CHF. For this reason, it is important to improve the healing of the scar in such a way that CHF development can be prevented. In recent years, more literature has emerged about Wnt/Frizzled signaling in infarct healing. In this review we will discuss the involvement of Wnt/Frizzled signaling in the different phases of infarct healing and possible therapeutic targets following MI.The Wnt/Frizzled signaling pathway is a transduction cascade of high complexity, which is fundamental for a wide range of physiological and developmental mechanisms, as well as disease states ,14. The The Wnt/Frizzled signaling pathway comprises of two major branches, the canonical (involving β-catenin) and the non-canonical pathways (independent of β-catenin), which share their main components: Wnts (the ligands) and Frizzleds (their corresponding receptors). Wnts are highly conserved glycoproteins (350 to 400 amino acids long) that are found in many organisms . NineteeModulation of Wnt signaling can occur through the action of several endogenous-secreted proteins. There are two distinct classes, the soluble frizzled-related protein (sFRP) class and the Dickkopf (DKK) class, which function in distinct manners. sFRPs modify Wnt/Frizzled signaling by scavenging Wnt proteins with a frizzled-like cysteine-rich domain and thereby intervene in the interaction between Wnt ligands and Frizzled receptors. The sFRP family also includes the Wnt inhibitory factor 1 (WIF-1) and Cerberus that function in similar ways. DKK proteins inhibit Wnt signaling by interacting with the LRP co-receptors. This causes internalization of the LRPs and thereby diminishing of the availability of co-receptors for Wnt signaling ,29.β-Catenin is the second messenger of the canonical Wnt signaling pathway and it is the most studied of all. In the absence of the ligand (Wnt), several factors including the adenomatous polyposis coli (APC), casein kinase 1 (CK1) and Axin activate glycogen synthase kinase (GSK) 3β. This in turn phosphorylates several Ser/Thr residues of β-catenin, which leads to the ubiquitination of the latter. Hence, β-catenin is degraded and cannot enter the nucleus in order to modulate gene transcription  pathway. For a more detailed description of these signaling pathways we refer to some competent reviews on this subject [The Wnt/Frizzled transduction cascade is able to signal via mechanisms that do not involve β-catenin. This can be accomplished via the Ca subject ,36.The ECM in the heart forms a three-dimensional structure, which serves as a framework for the attachment of myocytes in order to transmit the mechanical force. The cardiac fibroblast is the main cell type that is responsible for producing ECM components. By number, but not by volume, cardiac fibroblasts are the predominant cell type in the heart . In the + cells originated from BM cells compared to 3.4% in unbanded hearts [Traditionally, it was thought that myofibroblasts were only derived from resident cardiac fibroblasts. However, myofibroblasts can also be derived from other precursor cells -49. Therd hearts . Anotherd hearts , whereasd hearts . More prd hearts -49.et al. [In vitro experiments done by Carthy et al. also showed canonical signaling upon stimulation with Wnt3a in mouse fibroblasts [et al. [There is ample evidence suggesting that components of the Wnt/Frizzled pathway are upregulated in myofibroblasts following MI. Frizzled-2 (Fzd-2) and Dvl-1 have an increased expression four days after MI and this is located at the border zone of the infarct. In the course of time, this expression migrates toward the center of the infarct ,50. Prevet al. . In vitrroblasts . Wnt3a iroblasts . In addiroblasts , which croblasts . On the roblasts . In the roblasts . Additioroblasts , which i [et al. . On the  [et al. . Researc [et al. .Taken together, these findings support the hypothesis that differentiation of cardiac fibroblasts and the migrating properties of the myofibroblast can be modulated by Wnt/Frizzled signaling. However, it also emphasizes the complicacy of Wnt/Frizzled signaling in the induction of myofibroblast differentiation.et al. have extensively studied the role of FrzA  in cardiac repair [Barandon c repair ,55,56. Ic repair . The incc repair , which dc repair ,58. In ac repair .Subsequently, the role of FrzA on angiogenesis following MI was assessed. The capillary density in the scar was significantly higher in the FrzA Tg mice. Moreover, the vessel walls were more muscularized and the mean vessel lumen area was 3-fold higher compared to wild-type controls . FurtherIn vitro, recombinant sFRP-2 was shown to inhibit the procollagenase activity of bone morphogenic protein (BMP)-1 in primary cardiac fibroblasts, preventing the maturation of type I pro-collagen [sFRP-2 has also been shown to play a role in myocardial infarct healing ,62. In vcollagen . Followicollagen . Recombicollagen . This cocollagen . In addicollagen .et al. demonstrated contradictory results for sFRP-2 [et al. demonstrated that after MI, sFRP-2 injection in the infarct area improves cardiac function and is able to inhibit fibrosis and remodeling [et al. concluded that the same phenotype could be observed in sFRP-2 null mice [et al. monitored the infarct only up to two weeks [In contrast to these data, Kobayashi r sFRP-2 . It was r sFRP-2 . Followir sFRP-2 . So whermodeling , the latull mice . Even thwo weeks . At thissFRP-4 has been shown to play a profound role in infarct healing as well . In a MICollectively, these studies implicate that sFRPs play an important role in infarct healing and this is probably not only by the modulating effect between Wnt ligands and Frizzled receptors but also by other inhibitory or stimulating properties (for example on BMP-1).Recently, pyrvinium (an FDA-approved drug) has been described to possess Wnt signaling inhibiting properties . It poteIn our laboratory, we have developed a peptide fragment of Wnt5a (UM206) that occupies the binding sites for Wnt3a and Wnt5a on the Fzd-1 and −2 receptor, thereby preventing Wnt signaling  expression in both cell types and increased α-SMA expression in fibroblasts. Besides, it promotes cell cycle progression but only cell numbers of cardiac fibroblasts were increased [in vivo study was performed that had promising results [in vitro data, there was an increment in VEGF expression and capillary density [in vitro data, no information on α-SMA expression in the infarcted area was reported, which could give an indication for a correlation between cardiac function and the presence of myofibroblasts. Remarkably, another study showed more favorable results with β-catenin depletion instead of overexpression [Modifications in the expression of β-catenin can also serve as a tool to inhibit or promote Wnt/Frizzled signaling, since it is a downstream component of the cascade. ncreased . Based o results . In a ra density . These dpression . Here, cpression . This suin vivo MI studies, which demonstrate increased amount of blood vessels in the infarcted area when treated with the antagonist UM206 [Vascularization is important for maintenance of all tissues in the body. The importance of regulated Wnt signaling in vasculogenesis during early developmental stages has been shown in several studies with disruptions or mutations in the Wnt/Frizzled pathway. Deletion of Wnt-2 or Fzd-5, results in defects of the placental vasculature in mice ,70. Morest UM206  or overest UM206 .Taken together, the involvement of Wnt/Frizzled signaling in formation of new vessels during pathophysiological conditions is highly evident. Modulation in this signaling cascade following MI is a potential means of improving infarct healing.+ human CSCs have been shown to differentiate into myocytes, endothelial cells and smooth muscle cells in vitro. These differentiated myocytes were able to contract upon electrical stimulation [+ and Sca-1+ cells in the circulation [ex vivo[ex vivo expanded hematopoietic progenitor cells (HPCs) has been shown to decrease infarct size and improve EF, FS and LV end systolic diameter in mice following injection of these cells [ex vivo without losing multipotency [Until the recent discovery of cardiac stem cells (CSCs), the heart has been viewed as a terminally differentiated organ. These cardiac progenitor cells are multipotent and can give rise to new cardiomyocytes, smooth muscle cells and endothelial cells ,76. Hencmulation . Injectimulation . Recentlculation . These c [ex vivo, implicase cells , whereasse cells . Besidesipotency . Akt oveipotency . Moreoveipotency .Wnt signaling is necessary for maintenance of pluripotency , whereasThe wound-healing phase is a critical process for the clinical outcome of patients who have suffered from MI. Abnormal remodeling will result in a poorly healed scar that can cause dilatation of the left ventricle and ultimately progresses into CHF, which can be lethal. Since an increasing number of patients are affected by this problem, it is of major importance that therapeutic interventions do not just delay the progression, but rather prevent the development of CHF. The Wnt/Frizzled pathway is involved in many aspects of cardiac repair following MI and may be a promising therapeutic site for interventions. However, the precise targets and mechanism of intervention are still a matter of debate. Thus far, promising tools have been used in experimental settings, targeting Wnt/Frizzled signaling at several important sites involved in infarct healing such as (myo)fibroblasts, progenitor cells, neovascularization, inflammation, collagen maturation and several others that can contribute to an improved cardiac phenotype by minimizing the scar and reducing ventricular dilatation Figure. Wnt/FriThe discussed studies, in which interventions in Wnt signaling following MI are described Table, have diα-SMA = α-smooth muscle actin; APC = adenomatous polyposis coli; BM = bone marrow; BMP = bone morphogenetic protein; CHF = congestive heart failure; CK1 = casein kinase 1; CSC = cardiac stem cell; CVD = cardiovascular disease; DKK = Dickkopf; Dvl = Dishevelled; ECM = extracellular matrix; EF = ejection fraction; EMT = epithelial to mesenchymal transition; Endo-MT = endothelial to mesenchymal transition; Fzd-2 = Frizzled-2; FS = fractional shortening; GSK = glycogen synthase kinase; HPC = hematopoietic progenitor cell; JNK = c-jun N-terminal kinase; LRP = low-density lipoprotein receptor-related protein; LV = left ventricle; LVIDD = left ventricular internal diameter in diastole; MCS = mesenchymal stem cell; MI = myocardial infarction; MMP = matrix metalloproteinase; PMNL = polymorphonuclear leukocyte; sFRP = soluble frizzled-related protein; TCF/LEF = T-cell factor/lymphoid enhancer factor; Tg = transgenic; TGF = transforming growth factor; VEGF = vascular endothelial growth factor; WIF = Wnt inhibitory factor.The authors declare that they have no competing interests.KCMH carried out the bibliographic search, created the figures and wrote the draft. EPD contributed to this. WMB coordinated and helped with the draft of the manuscript. All authors read and approved the final manuscript.


Presence and origin of endemic foci of HTLV-2 infection in Africa remain a matter of debate.To better appreciate the epidemiological and molecular determinants of HTLV-2 infection in Central Africa, we performed a survey in 3903 inhabitants of a South Cameroon forest area, including 1051 Bakola Pygmies, 815 Baka Pygmies and 2037 Bantus living in their neighboring. HTLV-1 and HTLV-2 infection was determined by both specific serological (IFA and WB) and molecular (different generic and specific PCR) methods.HTLV-1/2 prevalence was of 3% (117/3903) with 90 HTLV-1 (2.3%) and 27 HTLV-2 (0.7%). Surprisingly, HTLV-2 infection was restricted to Bakola Pygmies (27/1051 2.5%) with no HTLV-2 infection in any of the 2852 Baka or Bantus individuals. In Bakola Pygmies, HTLV-2 seroprevalence increased with age, reaching 6.5% in the elder persons. Ongoing intrafamilial HTLV-2 transmission was evidenced. Lymphoid T cell lines  producing HTLV-2 antigens, were established from PBMCs cultures of HTLV-2 infected individuals. Sequences of a 672 nucleotide LTR fragment, obtained from 7 HTLV-2 samples, showed a very high degree of homologies among samples (< 1% nucleotide divergence) but also surprisingly with Amerindian HTLV-2 B strains. Complete sequence (8954 bp) of one isolate confirmed a typical HTLV-2 B strain.This study demonstrates clearly a HTLV-2 endemic population, with ongoing transmission, in Central Africa. Furthermore, it gives insights into several central questions regarding the origin and evolution rate of HTLV-2 and the migrations of infected populations.


These strains were likely introduced during ancient human population movements in Melanesia.Four of 391 Ni-Vanuatu women were infected with variants of human T-cell leukemia virus type 1 (HTLV-1) Melanesian subtype C. These strains had  Human T-cell leukemia virus type 1 (HTLV-1), a human oncoretrovirus, is the etiologic agent of adult T-cell leukemia and of tropical spastic paraparesis/HTLV-1–associated myelopathy. Molecular epidemiologic studies have shown HTLV-1 proviruses to be remarkably stable genetically. The low levels of genetic drift in this virus have been used as a means for monitoring viral transmission and the movement of ancient human populations . The fewPrevious reports have indicated that HTLV-1 is endemic in some remote or ancient populations in Melanesia –14. ThesGenetic characterization of the few available Melanesian HTLV-1 strains has indicated that these HTLV-1 strains are the most divergent, constituting molecular subtype C We carried out a large serologic and molecular study to determine the prevalence of HTLV-1 and associated diseases in the Vanuatu Archipelago. Vanuatu, formerly known as the New Hebrides, is a Y-shaped archipelago made up of ≈80 islands. It is located in Melanesia, in the South Pacific region, northeast of Australia and south of the Solomon Islands. Vanuatu has a population of ≈200,000 inhabitants, most of whom (95%) are of Melanesian origin and are known as the Ni-Vanuatu.ecade ago and were not based on stringent serologic criteria –14. No mIn February 2002, we recruited 391 women during a clinical survey for sexually transmitted diseases in various remote rural communities of western Ambae Island in the Penama Province of the Vanuatu Archipelago. Ambae Island, also known as Aoba, has a population of ≈9,500. The women participating in this survey were offered a complete clinical examination, with Papanicolaou test analysis for all women >25 years of age. For each participant, we obtained plasma and buffy coats from 5 mL of blood obtained by venipuncture. The blood samples were rapidly transferred to Institut Pasteur de Nouvelle-Calédonie, where plasma and buffy coats were isolated, frozen, and stored (at –80°C) until HTLV screening. Informed consent was obtained from each woman participating in the field survey. This study was approved by the Ministry of Health of the Republic of Vanuatu and was supported by the Vanuatu Family Health Association, a local nongovernmental organization. Samples were taken from 391 women  with the following stratification by age: 11.2% from women 15–24 years of age, 28.4% from women 25–34 years of age, 31.2% from women 35–44 years of age, 17.4% from women 45–54 years of age, and 11.8% from women ≥55 years of age.env-encoded glycoproteins: the HTLV-1–specific recombinant gp46-I peptide (MTA-1) and the specific HTLV-1/HTLV-2 recombinant GD 21 protein. Plasma samples were considered negative when no band were shown and indeterminate when partially reactive (Plasma HTLV-1 antibodies were detected by enzyme-linked immunosorbent assay (ELISA)  with Western blot  used for confirmation. On Western blot, plasma samples were considered HTLV-1–positive if they reacted to the 2 Gag proteins (p19 and p24) and both reactive ,16.gag-indeterminate profile . The 15 DNA samples studied were subjected to polymerase chain reaction with primers specific for the human β-globin gene to check that cellular DNA was amplifiable for all samples . We thenescribed . Fragmenescribed . Both thThe gp21 gene sequences of the 4 HTLV-1 strains involved were almost identical (99.6%–99.8 % nucleotide similarity) and were very similar to those of Melanesian strains. These strains were closely related (99.4%) to certain strains from Solomon Islanders  but were only 97.1%–98.3% similar to strains from Papua New Guinea residents  and from Australian aborigines (MSHR-1), respectively. Finally, the sequences of these new strains diverged from those of HTLV-1 strains from the 3 other main molecular subtypes  by 6% to 9%.The 4 new HTLV-1 LTR sequences were also very closely related (98%–100% nucleotide similarity). They displayed 2% nucleotide divergence from Mel 5 (from a Solomon Islander), the only available LTR from all the HTLV-1 subtype C strains. However, they also displayed up to 11% nucleotide divergence from HTLV-1 strains from other molecular subtypes.env and LTR HTLV-1 sequences from Melanesia, and on several representatives of HTLV-1 and STLV-1 strains from the various subtypes/subgroups as described (env analysis, which included 71 HTLV-1 strains (including 12 Melanesian strains and 1 from an Australian aborigine, Phylogenetic analyses were performed on all the available escribed , by the escribed . Analyseescribed . Indeed,In conclusion, we report, for the first time, the presence of HTLV-1 infection in a Ni-Vanuatu population living in remote villages. We also demonstrate that the viruses infecting these Ni-Vanuatu persons are novel HTLV-1 molecular variants belonging to the Melanesian divergent C subtype. This finding suggests that these viruses were introduced into Vanuatu by ancient migrations of Melanesian populations. The first people to reach Santa Cruz, Banks, Vanuatu, and the Loyalties Islands ≈3,600 years ago seem to have been Austronesian speakers . Epidemi


FAS and FASL genes are closely linked to the mechanism of the immune system and several polymorphisms in these genes have been associated with susceptibility to diseases. This study investigated the polymorphisms at positions -670 in the FAS gene, and -169 (IVS3nt169) and -124 (IVS2nt-124) in the FASL gene, among HTLV-1/2 infected subjects. Blood samples from 66 HTLV infected subjects and 192 seronegative individuals were collected, and polymorphisms were analyzed using a polymerase chain reaction (PCR) followed by RFLP analysis using restriction endonucleases. The products were visualized after electrophoresis in 4% agarose gel. The genotype frequencies of the FAS -670 polymorphism showed a higher prevalence of genotype -670GG among HTLV-1 infected subjects (46.97%) as compared to the control group . The comparative analysis between symptomatic and asymptomatic HTLV infected subjects showed a higher prevalence (p=0.0007) of the genotype -670GG among patients with TSP/HAM (50%) as compared to asymptomatic subjects (45%). TCD4+ and TCD8+ lymphocyte counts from HTLV infected and seronegative subjects were compared but no significant association was observed. The proviral load values according to the status of symptomatic and asymptomatic infection carrying different genotypes were compared but showed no statistical significance. The present results suggest that FAS -670 polymorphism seems to be associated with susceptibility to HTLV-1 and may increase the chance to develop TSP/HAM among HTLV-1 infected persons.