Primary cilia are present on almost all vertebrate cells, and they have diverse functions in distinct tissues. Cilia are important for sensation in multiple capacities in contexts as different as the retina, kidney, and inner ear. In addition to these roles, cilia play a critical part in various developmental processes. Of particular importance is the development of the neural tube, where cilia are essential for the transduction of the Sonic Hedgehog (Shh) signaling pathway that specifies neuronal cell fates. This relationship is well established and is the most recognizable function for cilia in the neural tube, but it may be part of a larger picture. Here, we discuss the links between cilia and Shh signaling, as well as suggesting additional roles for cilia, and mechanisms for their placement, in the neural tube. Since being functionally linked to the Sonic Hedgehog (Shh) signaling pathway, primary cilia have sparked enormous interest. The initial connection came from an unbiased forward genetic mouse screen in which a number of the mutations disrupted genes important for cilia; the resulting mutant embryos showed abnormal patterning of the neural tube,2. Shh sCore progress has been made towards understanding the mechanistic details behind the abnormal neural patterning of mouse mutants with disrupted cilia,2,14,15,Following the specification of the germ layers, the neural tube starts to form,27. The Patterning of the neural tube and correct cell fate specification are integral parts of proper development, and tight control of the Shh signaling pathway is required for appropriate ventral patterning and specification of motor neurons and interneurons,30. The In the ventral neural tube, a combination of the amount and duration of Shh signaling specifies six neural progenitor cell fates,30,40. DPtch1 and Gli1, is promoted.The transduction of the signal from Shh ligand to the Gli transcription factors is an intricate cascade involving many elements, including Patched (Ptch1), a transmembrane receptor, Smoothened (Smo), a membrane protein, Suppressor of Fused (SuFu), and Kif7, a possible motor protein. In the absence of Shh, Ptch1 inhibits Smo and causes repression of the pathway. This reDrosophila Costal-2, functions between Smo and the Gli proteins to both negatively and positively regulate the pathway[In vertebrate systems, this signaling cascade happens in the context of the cilium, and the dynamic transport of pathway members is key for proper transduction. When the pathway is off, Ptch1 is localized to the cilium whereas Smo is not; upon stimulation by Shh ligand, Ptch1 leaves the cilium while Smo becomes enriched there. The Gli pathway. Though Ift88 and Ift172, lead to either absent or severely shortened cilia; this results in no Shh signaling and a lack of both GliA and GliR. Thus, in Ift88pol and Ift172wim mutants, the lack of activation combined with derepression causes neural tube mispatterning[Cilia and Shh were initially linked through forward genetics in the mouse, since mutants defective in intraflagellar transport (IFT) proteins showed improper neural tube patterning, and IFT is necessary for ciliogenesis and cilia maintenance,2. The IIft122 null allele sister of open brain (sob)[Together, IFTA proteins and the cytoplasmic motor dynein power retrograde transport, which traffics proteins from the tip of the cilium back toward the cell. Unlike ain (sob),52. Thesain (sob),52.ling-ling), Ptch1, Smo, and Gli2 are all trafficked into the cilium, but no Shh transduction occurs, causing a dorsalized neural tube[Ift172avc1/+), rescues the patterning phenotype caused by the absence of retrograde transport[Ift122sopb/+ is also able to suppress the Dync2h1lln/lln phenotype[Ift122sopb/+ rescues the Dync2h1lln/lln phenotype suggests that Ift122 may also act outside retrograde transport, emphasizing the nuances of transport necessary for proper signaling. This is further highlighted by recent analysis of an allelic series of IFTA mutants, which demonstrated that proper Shh signaling relies on correct cilia architecture and protein trafficking[Though originally a topic of intense debate, the claim that transduction of the Shh pathway is intricately tied to proper cilia structure and transport has been supported by the identification of diverse mutants that all disrupt cilia structure or impair cilia protein transport. The simple ability to move pathway components into and out of the cilium is not enough for proper signal transduction; instead, the relative rates of anterograde and retrograde transport are critical. This is demonstrated most strikingly by the genetic interactions of anterograde and retrograde trafficking mutants. Dync2h1 is the heavy chain of dynein, the cytoplasmic motor responsible for retrograde transport, and when it is disrupted (as in its mutant ral tube,53. Interansport. Even mohenotype. Though fficking.In addition to the conserved cilia proteins critical in building and maintaining cilia through multiple phyla, other genes that are important for cilia also regulate Shh signaling. For example, Arl13b and Rab23 are small GTPases, and mutations in either lead to unique defects in neural tube patterning. Strikingly, this is due to loss of Arl13b preventing full activation of GliA but leaving normal GliR function intact. In geneAlthough the intricate connections between cilia and Shh signal transduction are the best understood at present, the cilia within the neural tube may have additional functions. Cilia on cells that line the ventricular zone of the neural tube are aligned and extend into the lumen. This organization is most obvious at the most ventral levels of the ventricular zone near the floor plate. It seems unlikely that this specific orientation is an artifact but rather suggests that the placement of these cilia may be important to their function, to their ability to respond to signal, or to the morphology of the neural tube. For example, it is not yet known whether cilia in the neural tube function solely to transduce the Shh or if they actually sense the Shh ligand. Perhaps this organization of cilia in the ventral neural tube is most obvious as their length there is proportional to their ability to detect Shh ligand. On the other hand, the position of the cilium directly relates to the plane of cell division. As the cells lining the lumen are highly proliferative progenitor cells, a subset of which remain progenitors, the positioning of the cilium could be critical for cellular asymmetry. Shh has a well established role as a mitogen in other tissues,64, and CE is critical to neural tube formation as it elongates the tissue so that the brain and spinal cord eventually form. Cells that participate in CE must be polarized, and this specific cell shape is dependent on the PCP cascade,28,65,66Inturned or Fuzzy, which regulate PCP signaling and CE, also display abnormal ciliogenesis and secondary Shh defects[Analyses of mutants in either PCP signaling or ciliogenesis indicate that neither PCP signaling nor CE require cilia. Mutants with disrupted cilia display normal CE, and cilia play no known role in the process. Despite this, a variety of data link PCP signaling and cilia-20, rais defects,21. Toge defects. Further defects. TogetheGiven the role of PCP in cilia orientation, it is vital to determine whether or not PCP signaling influences the position and/or organization of cilia in the ventricular zone of the neural tube. The first step would be to examine cilia in the neural tube of known PCP mutants to determine if the anecdotal arrangement of cilia is disturbed. Finding a way to disrupt the placement of cilia will confirm that their position is indeed deliberate, and from there we will be able to pose the question of mechanism and function.Caenorhabditis elegans nervous system is ciliated, and those cilia are critical for chemosensation[What remains unclear is whether the cilia in the neural tube are there solely to transduce Shh signaling. After all, in the lungs, motile cilia that clear mucus also express taste receptors, so perhensation. In the ensation,25. Addiensation-24. FurtSince a mouse mutant linked Shh and cilia almost a decade ago, the field has focused on understanding the mechanistic links between the two. This work has painted a dynamic picture, whereby key components move into and out of cilia to regulate the ratio of GliA and GliR and ultimately specify distinct cell fates in the neural tube. Other signaling pathways have also been linked to cilia, although whether such links are critical in the neural tube remains unclear. The projection of cilia into the lumen of the ventricular zone strongly suggests that they could play a sensory role, in which case the positioning of these cilia would be crucial and highly regulated, possibly through PCP signaling. But the biggest question remains: do these cilia play another functional role? Cilia in other contexts are strikingly suggestive, and the field is poised to investigate for what other tasks, if any, the cilia of the neural tube may be responsible.A-P: anterior-posterior; BBS: Bardet-Biedl syndrome; CE: convergent extension; CSF: cerebral spinal fluid; D-V: dorsal-ventral; IFT: intraflagellar transport; NTD: neural tube defect; PCP: planar cell polarity; Ptch1: Patched; Sh: Sonic Hedgehog; Smo: Smoothened; SuFu: Suppressor of Fused.The authors declare that they have no competing interests.SNB and TC wrote the manuscript. Neither has any conflict of interest to report. Both authors read and approved the final manuscript.


ATL is a highly aggressive leukemia/lymphoma which was first proposed as a new disease entity in 1977. The long clinical latency and low incidence of ATL indicate that some genetic changes are involved in malignant transformation and monoclonal expansion of HTLV-1-infected cells. Monoclonal proliferation of HTLV-1-infected cells is observed in a part of the virus carriers, who are considered to be the high risk group for development of ATL [We reviewed the process of senescence and how proteins allow the monoclonal proliferation of ATL cells.Constitutive activation of STAT as well as functional impairment and stabilization of p53 protein found in the PBMCs of ATL patients are supposed to be one base for ATL development ,3. In adFurthermore, the soluble form of CD30 (sCD30) is elevated in serum of ATL patients and correlates with the aggressiveness of ATL  (Our obsNot only the inhibition of senescence by impaired function or constitutive activation of proteins in ATL cells, but also immune regulation by soluble proteins in serum of HTLV-1-infected patients must be required for the progression of ATL.


The small GTPase ADP ribosylation factor 6 (ARF6) mediates endocytosis and has in addition been shown to regulate neuron differentiation. Here we investigated whether ARF6 promotes differentiation of Neuro-2a neuronal cells by modifying the cellular lipid composition. We showed that knockdown of ARF6 by siRNA in Neuro-2a cells increased neuronal outgrowth as expected. ARF6 knockdown also resulted in increased glucosylceramide levels and decreased sphingomyelin levels, but did not affect the levels of ceramide or phospholipids. We speculated that the ARF6 knockdown-induced increase in glucosylceramide was caused by an effect on glucosylceramide synthase and, in agreement, showed that ARF6 knockdown increased the mRNA levels and activity of glucosylceramide synthase. Finally, we showed that incubation of Neuro-2a cells with the glucosylceramide synthase inhibitor D-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol (D-PDMP) normalized the increased neuronal outgrowth induced by ARF6 knockdown. Our results thus show that ARF6 regulates neuronal differentiation through an effect on glucosylceramide synthase and glucosylceramide levels. Neuron development and differentiation are complex processes that involve dynamic cell morphology changes. It has been suggested that endosomal trafficking is crucial for actin cytoskeleton structure and neuronal cell differentiation Endocytosis is also regulated by modulation of the lipid composition of cellular membranes In this study, we investigated whether ARF6-dependent neuron differentiation is regulated by alterations in lipid composition. We found that ARF6 knockdown resulted in increased glucosylceramide content and glucosylceramide synthase activity in Neuro-2a neuronal cells. Furthermore, we found that ARF6-dependent neuron differentiation is regulated by the altered glucosylceramide synthase activity in neuronal cells.Neuro-2a cells were purchased from the American Type Culture Collection . Cells were cultured in DMEM with 10% serum and were differentiated in medium without serum.Total RNA was extracted with an RNeasy Kit , and cDNA was synthesized with the high-capacity cDNA Reverse Transcription Kit . mRNA expression of genes of interest was analyzed with TaqMan real-time polymerase chain reaction in an ABI Prism 7900 HT Detection System (Applied Biosystems) and normalized to β-actin. The following TaqMan Gene Expression assays from Applied Biosystems were used: GCS (Ugcg) Mm00495925_m1, beta-Actin Mm01205647_g1, NeuroD1 Mm01946604_s1, Rbfox3 Mm01248771_m1 and 36B4 Mm007725448_s1.AUCUGACAUUUGACACGAATT, antisense UUCGUGUCAAUGUGAGAUCA; siARF6-2: sense GCAAGACAACGAUCCUGUATT, antisense UACAGGAUCGUUGUCUUGCCG).Neuro-2a cells were cultured at 30% confluence and transfected with 100 pmol/l target-specific or scrambled control siRNA (50 nmol/l) using Lipofectamine RNAiMAX  according to the manufacturer's protocol. The siRNAs used were from Applied Biosystems (siARF6-1: sense Plasmids for ARF6-cyan fluorescent protein (CFP), T27N ARF6-CFP and Q67L ARF6-CFP were purchased from Addgene . Neuro-2a cells were transfected with plasmids at 70–80% confluence using jetPRIME  according to the manufacturer's instruction. Transfection efficiency (determined by CFP by using fluorescent microscopy) was around 50%.The neuronal outgrowth was assessed in Neuro-2a cells that were cultured in DMEM with 10% serum. Long outgrowths from Neuro-2a cells were defined as having a length at least double that of the cell diameter. Randomly selected micrographs of 100–300 cells from 4 different experiments were analyzed for percentage of cells with outgrowth or long outgrowth.Lipids were extracted from Neuro-2a cells using the Folch procedure Phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine and sphingomyelin were quantified as described Ceramide and glucosylceramide were analyzed using straight-phase HPLC coupled to a Quattro Premier XE triple quadrupole mass spectrometer . The lipids were separated using a Sunfire 150×2.1 silica column with 3 µm particles (Waters). The mobile phase A was isohexane∶isopropanol (95∶5) and the mobile phase B was isohexane∶isopropanol:50 mmol/l ammonium formate in water (25∶65∶10). The gradient went from 100% A (held for 1 min) to 100% B in 5 min. After 4 min at 100% B, the gradient returned to 100% A and the column was equilibrated for 3 min. Thus, the total runtime was 13 min. The flow rate was 500 µl/min. A postcolumn flow of methanol∶isopropanol (1∶1) at 100 µl/min was used to ensure optimal ionization of the lipids in the ion source. Samples were injected in mobile phase A. Ceramide and glucosylceramide were detected using multiple reaction monitoring, quantified using external standards and normalized against their respective internal standard and the cellular protein content.N- aminocaproyl-sphingosine (NBD C6-ceramide) to NBD C6-glucosylceramide in cells, as described The activity of glucosylceramide synthase was measured by following the conversion of fluorescently labeled ceramide 6-t test or one-way ANOVA with Dunnett's post-hoc test.Data are presented as mean ± s.e.m. Statistical significance was determined with Student's It has previously been observed that inactivation of ARF6 by overexpression of a GAP or a dominant-negative ARF6 promotes neuron differentiation, as shown by increased neuronal outgrowth, in various neuronal cell systems In this study, we used Neuro-2a neuronal cells and knocked down ARF6 with siRNA to study the effects on neuron differentiation . We founTo investigate whether ARF6 knockdown affects the lipid composition of Neuro-2a cells, we performed a lipidomics analysis of Neuro-2a cells transfected with control or ARF6 siRNA. Interestingly, we showed that ARF6 knockdown resulted in significantly increased levels of glucosylceramide and significantly decreased levels of sphingomyelin but did not change ceramide levels . Levels Our results indicate that ARF6 is a regulator of the sphingolipid composition of Neuro-2A cells. Numerous studies have shown that glucosphingolipids, especially gangliosides, are important for neuron differentiation and development. The ganglioside GM1, for example, has been shown to promote neuron outgrowth in both primary neurons and neuronal cell lines Glucosylceramide and sphingomyelin are synthesized from ceramide in the Golgi apparatus  [26], [2Increased glucosylceramide synthase activity will promote the conversion of ceramide to glucosylceramide. Our results thus indicate that glucosylceramide synthase activity is important for the ARF6-regulated modification of sphingolipid levels in Neuro-2a cells.Finally, we tested if ARF6 regulates differentiation of Neuro-2a cells through an effect on glucosylceramide synthase. We knocked down ARF6 for 48 h to stimulate neuronal outgrowth and then incubated the cells in the presence or absence of the glucosylceramide synthase inhibitor D-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol (D-PDMP) for 20 h. As expected, the number of cells with long outgrowths increased significantly following ARF6 knockdown . InteresThe lipid composition of neuronal membranes has been shown to be important for actin structure and endosomal trafficking In conclusion, our results indicate that ARF6 regulates neuronal differentiation through modulation of glucosylceramide levels.


These two transcription factors, PPARγ and Runx2, are generally regarded as the master regulators of adipogenesis and osteogenesis. This review will summarize signaling pathways that govern MSC fate towards osteogenic or adipocytic differentiation. A number of signaling pathways follow the inverse balance between osteogenic and adipogenic differentiation and are generally proosteogenic/antiadipogenic stimuli. These include β-catenin dependent Wnt signaling, Hedgehog signaling, and NELL-1 signaling. However, other signaling pathways exhibit more context-dependent effects on adipogenic and osteogenic differentiation. These include bone morphogenic protein (BMP) signaling and insulin growth factor (IGF) signaling, which display both proosteogenic and proadipogenic effects. In summary, understanding those factors that govern osteogenic versus adipogenic MSC differentiation has significant implications in diverse areas of human health, from obesity to osteoporosis to regenerative medicine.Mesenchymal stem cells (MSC) are multipotent cells, functioning as precursors to a variety of cell types including adipocytes, osteoblasts, and chondrocytes. Between osteogenic and adipogenic lineage commitment and differentiation, a theoretical inverse relationship exists, such that differentiation towards an osteoblast phenotype occurs at the expense of an adipocytic phenotype. This balance is regulated by numerous, intersecting signaling pathways that converge on the regulation of two main transcription factors: peroxisome proliferator-activated receptor- Mesenchymal stem cells (MSC) are multipotent stromal cells capable of self-renewal and capable of multilineage mesenchymal differentiation . These nMSC derived from bone marrow (BMSC) are relatively scarce in number but like all MSC have a capacity for repeated culture expansion while retaining their growth potential and multipotency . BMSC tyγ (PPARγ) transcriptional activity , 3029, 3ce versa –34. Sevece versa , 36. Sevgnaling) , 38, Hedgnaling) , 40, andgnaling) , 42. Disgnaling) , 44. Whignaling) , 46, BMPgnaling) , 48. Insγ and Runx2. PPARγ is generally considered the master regulator of adipogenesis and also has well-described anti-osteoblastogenic effects. Likewise, Runx2 is regarded as the master regulator of osteogenesis. Together, they are in large part responsible for mediating the effects of various cytokines in determination of adipogenic versus osteogenic MSC differentiation. Typically, increased expression of one transcription factor is associated with downregulation of the other [γ. For example, Osterix and CCAAT/enhancer-binding family of proteins (C/EBP) play important adjunctive roles , a cotranscription factor [Runt, is homologous to the Runt sequence in Drosophila. Members of the Runx family have various roles in determining stem cell commitment; Runx1 determines hematopoietic stem cell differentiation [β1), BMP, Wingless type (Wnt), Hedgehog (HH), and (Nel)-like protein type 1 (NELL-1) [Runx2−/−) have an absence of differentiated osteoblasts and bone and die shortly after birth [Runx2 null phenotypes cannot be rescued by the overexpression of other osteogenic factors, although the cleidocranial dysplasia-like phenotype of Runx2+/− mice can be partially rescued [Originally identified as the binding site for polyomavirus enhancer binding protein (PEBP), Runx was later identified as the Moloney murine leukemia virus enhancer core binding protein . The Run subunit , 57. In n factor . The DNAntiation , Runx2 dntiation , and Runntiation , 61. Runntiation , 63. Retntiation . Howeverntiation –68. As t(NELL-1) –71. Miceer birth . Such Ru rescued , 74. Whiα [δ and PPARγ were since discovered. All three PPARs are found in mammals and are activated by polyunsaturated fatty acids [α and PPARδ are expressed during adipogenesis, PPARγ is adipocyte restricted and more rapidly increases in expression during early adipogenesis [γ is expressed during adipogenesis as two isoforms, PPARγ1 and PPARγ2, the latter being predominant in adipose tissue [γ1 is expressed at lower levels in adipose tissue among other tissues, including breast and prostatic tissue [γ is principally regarded as the master regulator of adipogenesis, for no other factor can rescue adipocyte formation in the event of PPARγ knockout, and generally all proadipogenic cell signaling pathways converge with PPARγ [Peroxisome proliferator-activated receptors are members of the steroid/thyroid hormone receptor gene superfamily . Initialα , subsequty acids , interacty acids . While bogenesis , 80. PPAe tissue . PPARγ1 c tissue –83. PPARth PPARγ .γ must occur for any proadipogenic effects. Even then, the ligand is only necessary in the commitment phase for the adipocyte lineage, whereas PPARγ expression is necessary for both commitment and differentiation phases [γ activation through exposure to an exogenous ligand. By contrast, preadipocytes were able to continue with adipogenic differentiation without exposure to ligand [γ is thiazolidinediones (TZDs), which are potent PPARγ agonist among several other derivatives of polyunsaturated acids [γ, although induced adipogenesis [γ without functional ligand-binding domains was able to support adipocyte differentiation [γ ligand activation.It is currently believed that a ligand-dependent activation of PPARn phases , 85. Oneo ligand . One suced acids . Recentlogenesis , 85. Morntiation , which iγ in mice have confirmed its central role in adipogenic differentiation. Cells derived from PPARγ+/− mice demonstrate a reduced ability to differentiate into adipocytes [γ-deficient embryonic stem cells fail to differentiate into adipocytes and instead differentiate into osteoblasts. Additionally, PPARγ+/− mice have demonstrated increased bone mass with increased osteoblastogenesis, while having a marked decrease in fat stores [γ2 have decreased expression of both PPARγ1 and PPARγ2 in white adipose tissue, while exhibiting increased bone formation [γ in murine adipose tissue led to a loss of both brown and white adipocytes [Studies from genetic manipulation of PPARipocytes . PPARγ-dt stores . Similarormation . In anotipocytes .γ. Several PPARγ agonists/ligands, namely, TZD rosiglitazone and 15-deoxy-delta -PGJ2, promote BMSC adipogenesis while inhibiting osteogenesis [in vivo, where chronic treatment of mice with low-affinity TZD troglitazone induces increased bone marrow adipocytes, without affecting bone mass [in vitro in a PPARγ2-dependent manner [In vivo, neglitazone did not demonstrate an effect on bone, with unaffected expression levels of Runx2 [There is much evidence supporting the anti-osteoblastogenic and proadipogenic properties of PPARogenesis , 89. Howogenesis . A similone mass . Conversone mass , 91. Thiβ-catenin dependent) and noncanonical (β-catenin independent) pathways  signaling has been identified to play an essential role in cell fate determination, proliferation, and differentiation , 93. Dyspathways .β-catenin dependent pathway initiates with the binding of extracellular Wnt ligands to the seven-pass transmembrane frizzled receptors (Frz) expressed at the cell surface [β-catenin, promoting its degradation. Wnt stimulation inhibits the Axin/GSK3/APC complex, and β-catenin accumulates rather than being degraded, and levels of nuclear β-catenin increase. Once inside the nucleus, β-catenin can heterodimerize with lymphoid enhancer-binding factor/T cell factor [β-catenin dependent Wnt signaling elicits gene transcriptional activity to influence MSC lineage determination [β-catenin independent manner [The  surface . This in surface . The resl factor . Ultimatmination   protein family has been identified in all vertebrates and classified into three structural homologues: Sonic Hedgehog (SHH), Indian Hedgehog (IHH), and Desert Hedgehog (DHH). DHH expression is typically limited to male reproductive tract  and willAll three HH morphogens follow the same, highly conserved HH signaling pathway . First, α and PPARγ [in vitro studies evaluating RNAi scans on Drosophila genome have confirmed the antiadipogenic function of HH signaling. Specifically, HH signaling blocked differentiation of white adipocytes. Likewise, transgenic activation of HH signaling in both Drosophila and mammalian models impaired fat formation [The antiadipogenic potential of HH signaling in MSC has been observed across a variety of adipocyte and multipotent cell lineages. Generally, adipogenesis in MSC, as it relates to HH signaling, occurs as a result of decreased Gli1, Gli2, Gli3, and PTCH expression . Conversnd PPARγ . Moreoveormation , 141. Usormation .in vivo and in vitro data suggest that bone formation occurs via a positive feedback loop. That is, HH-induced osteoblastogenesis requires BMP signaling, and together they elicit a synergistic expression of alkaline phosphatase activity [In addition to its antiadipogenic properties, HH signaling is well known to stimulate MSC osteogenic differentiation. While the exact mechanism and stage at which HH acts during osteoblastogenesis are not completely understood, both activity . This poactivity . In the activity –147. In activity . It is iactivity , 147. Thin vitro in primary cultures of fetal rat calvarial cells and MC3T3 cell line cultures [The secreted molecule NELL-1 (NEL-like protein 1) was first discovered to have osteoinductive properties by its overexpression during premature bone formation in human sporadic coronal craniosynostosis , 150. NEcultures . Moreovecultures . Thus, Ncultures .in vivo NELL-1 administration induces significant calvarial defect healing in rats [In vivo, several studies have demonstrated that NELL-1 has comparable bone regeneration capacity as BMP-2, in both calvarial defect and spinal fusion models, among others [β-TCP scaffolds [in vivo models, ranging from rodents to large preclinical animals [The osteoblastogenic effects of NELL-1 have been studied in the context of bone tissue engineering. For example,  in rats . When NE in rats . In vivog others , 155. NEg others . A varieg others , 157, uscaffolds . In a sh animals .Runx2 deficient mice, overexpression of NELL-1 was not sufficient to rescue mineralization, whereas absence of NELL-1 significantly decreased Runx2 activity in vitro [β1 was recently identified as the first cell surface receptor of NELL-1 [β1 expression [β1 blocked at least some of the cellular effects of NELL-1, including induction of pre-osteoblast attachment [Mechanistically, NELL-1 is directly regulated by the transcription factor Runx2 , 154. NEin vitro . Integrif NELL-1 . Cell supression . Moreovetachment . NELL-1 tachment , 161  superfamily, are extracellular cytokines originally isolated from bone extract and found to induce of ectopic chondrogenesis and osteogenesis [Bone morphogenetic proteins (BMPs), members of transforming growth factor-ogenesis . BMPs arogenesis . Over 20ogenesis , 166. Boogenesis , 168.BMPs produce their effects through interaction with two serine-threonine kinase cell surface BMP receptors (BMPRs). Type II BMPRs initiate signaling upon binding to a BMP ligand, following which recruitment, phosphorylation, and activation of type I BMPRs occurs , 170. Whγ via zinc finger transcription factor Schnurri-2 and C/EBPα, which exhibit synergistic, adipogenic effects [γ signaling and BMP-associated adipogenesis [γ activation and adipogenic differentiation [γ expression and BMP-associated adipogenesis [BMP induced adipogenesis involves both Smad1/5/8 and MAPK activation . BMP ind effects , 174. Acogenesis . Similarntiation . Conversogenesis . Investiogenesis , 176. Whogenesis , 176. BMogenesis , 178. Foogenesis . Moreoveogenesis , 180.in vitro and in vivo [BMP signaling is one of the central signaling pathways involved in the induction of osteogenic differentiation and regulation of bone formation. Multiple murine studies involving genetically modified BMP ligands, BMP receptors, and BMP inhibitors demonstrate a critical role for BMP signaling in bone formation –184. For in vivo –197. Fur in vivo . It is i in vivo . Further in vivo , 201.The precise determinants that govern BMP signaling induced adipogenesis versus osteogenesis in MSC are not well understood. Two variables that may determine the effects of BMP on MSC differentiation have been observed: dosage and receptor type. In terms of dosage, lower concentrations of BMP-2 have been shown to directs towards adipocyte formation, while higher concentrations favor osteogenic differentiation in C3H10T1/2 . HoweverDiscovered over fifty years ago, insulin-like growth factor-I (IGF-I) was originally identified as a soluble factor with insulin-like properties and induced by a growth hormone. Since then, we have developed a better understanding of this cytokine, especially in regard to its contribution towards bone formation and remodeling , 204 andIGF-1 produces its effect by inducing several intracellular signaling pathways. IGF-1 first binds to the IGF-1 receptor, which autophosphorylates the receptor intracellularly at the kinase domain. With the receptor now activated, various protein substrates are consequently activated, including insulin receptor substrate-1 (IRS-1) and Src homology and collagen protein (SHC) . IRS-1 gDuring bone remodeling, IGF-1 is released from the bone matrix to stimulate MSC osteoblastogenesis via activation of mammalian target of rapamycin (mTOR). This allows for the maintenance of both bone structure and mass, both of which were downregulated in mice with knockout of IGF-1 receptors in pre-osteoblastic cells . Similarβ1 led to accelerated bone healing [Combination of IGF-1 with various other growth factors provides additional insight on the mechanism of bone formation by IGF-1. For example, the addition of PDGF with IGF has been demonstrated to be more efficacious than either alone in terms of osteogenic induction in ASC . Likewis healing . Another healing . PTH is  healing , with a  healing . Further healing .γ, the key regulator for adipogenesis. Thus, a critical threshold of Akt activity, as regulated by IGF-1, contributes to the maintenance of cell proliferation, growth, and adipogenic differentiation [Interestingly, IGF-1 has been found to promote both adipogenic and osteogenic differentiation. For example, IGF-1 induces cell division of adipocyte precursor cells . In addintiation .γ or Runx2 expression, transcriptional activity, or both. Although the mechanisms have not been fully discerned, many of these growth factors tend to elicit an “inverse relationship” between adipogenic and osteogenic differentiation. As discussed, Wnt, HH, and NELL-1 signaling follow this pattern, exhibiting proosteogenic/antiadipogenic effects [β1 [Numerous signaling pathways induce the adipogenic and/or osteogenic differentiation of MSC, not all of which were covered in this review. The majority of signaling pathways ultimately converge downstream affecting PPAR effects . Other w effects , TGF-β1 ects [β1 , 224, anects [β1 , to nameects [β1 . Howeverects [β1 , 227–229


High plasma uric acid (UA) is a prerequisite for gout and is also associated with the metabolic syndrome and its components and consequently risk factors for cardiovascular diseases. Hence, the management of UA serum concentrations would be essential for the treatment and/or prevention of human diseases and, to that end, it is necessary to know what the main factors that control the uricemia increase. The aim of this study was to evaluate the main factors associated with higher uricemia values analyzing diet, body composition and biochemical markers.415 both gender individuals aged 21 to 82 years who participated in a lifestyle modification project were studied. Anthropometric evaluation consisted of weight and height measurements with later BMI estimation. Waist circumference was also measured. The muscle mass (Muscle Mass Index – MMI) and fat percentage were measured by bioimpedance. Dietary intake was estimated by 24-hour recalls with later quantification of the servings on the Brazilian food pyramid and the Healthy Eating Index. Uric acid, glucose, triglycerides (TG), total cholesterol, urea, creatinine, gamma-GT, albumin and calcium and HDL-c were quantified in serum by the dry-chemistry method. LDL-c was estimated by the Friedewald equation and ultrasensitive C-reactive protein (CRP) by the immunochemiluminiscence method. Statistical analysis was performed by the SAS software package, version 9.1. Linear regression (odds ratio) was performed with a 95% confidence interval (CI) in order to observe the odds ratio for presenting UA above the last quartile (♂UA > 6.5 mg/dL and ♀ UA > 5 mg/dL). The level of significance adopted was lower than 5%.2 OR = 2.28(1.13-4.6) and lower MMI OR = 13.4 (5.21-34.56) showed greater chances of high UA levels even after all adjustments . As regards biochemical markers, higher triglycerides OR = 2.76 (1.55-4.90), US-CRP OR = 2.77 (1.07-7.21) and urea OR = 2.53 (1.19-5.41) were associated with greater chances of high UA . No association was found between diet and UA.Individuals with BMI ≥ 25 kg/mThe main factors associated with UA increase were altered BMI (overweight and obesity), muscle hypotrophy (MMI), higher levels of urea, triglycerides, and CRP. No dietary components were found among uricemia predictors. Uric acid (UA) is a waste product of the human purine balance. It is formed by adenosine, inosine, hypoxanthine, adenine and guanine . AdditioHigh urate concentrations are associated with articular deposits and the onset of (gout) arthritis , metabolOther confounding factor is the diet and the relationship between diet and UA has not been fully elucidated, since most studies do not measure urate basal concentrations, do not exclude confounding factors and do not correctly evaluate ingested nutrients . HoweverSome confounding factors can influence the hyperuricemia because several variables are associated with each other -19, whatUric acid may be beneficial (antioxidant) or deleterious in high concentrations. Hence, the manegement of UA serum concentrations is promising in the treatment of certain diseases . TherefoHence, the present study aimed to evaluate what are the main factors associated with higher uricemia values, analyzing diet, body composition and biochemical markers.Mexa-se Pró-Saúde [Move for Health]” from 2002 to 2006. This program is offered to patients with non-communicable chronic diseases and consists of regular physical exercise and nutritional counseling. The Center for Physical and Nutritional Metabolism (CeMENutri) has conducted this program in Botucatu since 1992. Botucatu is a city located in mid-São Paulo State, approximately 230 km west of the capital city. It has a population of 121,274 habitants [A descriptive cross-sectional study was conducted in a subgroup of participants clinically screened for the lifestyle modification program “abitants .The inclusion criteria for participants were individuals of both genders with at without metabolic or motor disabilities that would limit physical exercise.A convenience sample was consisted of 1,075 individuals who were 51.6 ± 10.2 years old, and 59% of them were females. All the subjects signed a free-consent form, and the research project was approved by the Research Ethics Committee (document no. CEP 3272–2009) of the Botucatu School of Medicine (FMB), Univ Estadual Paulista - UNESP, Brazil. Of the 1,075 subjects, 415 had biochemical, anthropometric and dietetic data.Dietary intake data was determined by using 24-hour recalls. The diet was documented by trained professionals, and in order to obtain accurate information, the subjects were asked how often they usually ate during the day, what food varieties were consumed, how food was prepared, what the serving size was, and what food/meal brands were consumed. The diets were analyzed by NutWin® software (2002), version 1.5 , and theBody weight was measured by a platform-type anthropometric scale (Filizola®) with a maximum capacity of 150 kg and an accuracy of 0.1 kg. Height was determined by a portable Seca® stadiometer with accuracy of 0.1 cm . By usinWaist circumference (WC) was measured at the point midway between the last rib and the iliac crest. A steel Sanny® anthropometric tape measure (without a lock) was used for all measurements.A bioelectrical impedance device  was used to determine body fat percentage (% body fat)  and bodySystolic and diastolic arterial blood pressure was evaluated with the individual in the seated position according to the procedures described by the VI Brazilian Guidelines on Arterial Hypertension . Values Blood samples were collected after overnight fasting for 10 to 12 hours using a vacuum venous puncture. The individuals were previously instructed to not perform vigorous physical exercise 24 hours and/or not drink alcohol 72 hours before collection. Laboratory analyses of lipid parameters , glucose, uric acid, urea, gamma-glutamyl transferase (γ-GT), albumin and total proteins were performed within 4 hours after blood collection using the Dry- Chemistry method  while C-reactive protein (CRP) was measured by a high-sensitive Chemiluminescent method (Siemens Diagnostics), at the Clinical Analyses Laboratory of the Botucatu School of Medicine University Hospital - UNESP in Botucatu/SP. Low density lipoprotein (LDL-c) was calculated by Friedewald equation (LDL-c = TC- (HDL-c + TG/5) . GlomeruDiagnosis of the metabolic syndrome was performed according to the NCEP-ATP III criteria ,33 with th quartile . Overweight was classified as BMI ≥ 25 kg/m2[th quartile).Higher uric acid was considered when was higher than the 4 25 kg/m2, altered 25 kg/m2,33. Low  25 kg/m2. Hypertr 25 kg/m2,33, lowe 25 kg/m2,33 and h 25 kg/m2. Higher  25 kg/m2, higher p value < 0.05 was adopted as significant.Statistical analyses were conducted by using the SAS software for windows  and STATISTICA 6.0. Sample normality was tested by the Shapiro-Wilk test. Descriptive statistics were performed for the study, and continuous variables are presented as means ± standard deviation (SD) or median and interquartile intervals. UA values were divided into quartiles for both genders, and one-way ANOVA and generalized linear model considering gamma distribution and link function log were performed in order to observe the difference between variables according to the increase in the UA quartile. Logistic regression (Odds ratio) was made in order to determine the probability of UA increase (♂UA > 6.5 mg/dL and ♀ UA > 5 mg/dL) by food intake, anthropometry and biochemical analysis. Backward stepwise multiple regression analysis was used to determine the main components responsible for UA increase according to gender. A Higher UA concentrations (last quartile) were associated with higher values for body adiposity markers  whilst lower UA concentrations Q1) were associated with higher MMI values when compared to the other quartiles. There was no difference in relation to the individuals’ age or height  showed higher chances of high uric acid. When MS was adjusted for BMI, % body fat and MMI (Model 2) remained significant, and WC analysis was not performed, since it is part of the MS criteria. The four body-composition components evaluated remained significant when MS was removed from the adjustment and CRP (inflammation) gamma-gt LDL-c, glomerular filtration (creatinine and urea) were added. When the MS components  were added to the adjustment, only high BMI and low muscle mass remained significant (Table The odds ratio for the UA at the last quartile (♂ UA > 6.5 mg/dL and ♀ UA > 5 mg/dL) was performed, according to alterations in body composition. In model 1, additionally to the adjustment for gender and age, all body composition components were included in the same model, because it was possible to observe the influence of each compartment alone. Individuals with BMI, WC and % body fat above normality as well as with little muscle mass MMI < P5 showed As regards dietary intake, intake >60% of carbohydrate of total caloric intake when compared to <50% of CHO, represented higher chances of UA increase (Model 1). It is known that high CHO intake can increase glycemia and/or TG, and these may be related to UA increase. Hence, both were adjusted (Models 2 and 3). When high CHO intake was adjusted for glycemia or TG, the effect of CHO on UA was lost. Thus CHO intake could directly influence UA serum concentrations, firstly increasing glycemia or TG  was added. The altered values for creatinine, HDL-c, LDL-c and TG, conjointly with values in the last quartile for gamma-gt and CRP were significant in increasing the chances of individuals’ showing high UA. When body composition was added to the adjustment (Model 2), the creatinine and HDL-c values lost their effect; however, gamma-gt, LDL-c and TG remained significant. MS and glomerular filtration rate were also added to the adjustment (Model 3); TG remained significant, and the last CRP quartile and altered urea values became significant, thus increasing the chances of UA increase. Hypertension and glomerular filtration rate were added to the model, and MS was removed Model 4) because, in this way, all MS components would be gathered in the same adjustment model, and no modifications were observed in relation to the latter model, that is, altered urea and TG values; and values above the CRP last quartile were associated with UA increase ; and HDL-c (negatively) were associated for women.et al., 2008 [et al. (2007) investigated UA concentrations in a population consisting of black and white adults for ten years and identified an independent relationship of creatinine and urate [UA serum concentrations are maintained through the synthesis and excretion of urate ; hence, l., 2008  reportednd urate .Creatinine serum concentration was one of the main UA-related factors for females. In the odds-ratio model Table , alteredThe increase in uricemia distribution quartiles did not follow age increase. It is usually reported that, as age advances, a gradual reduction in glomerular filtration occurs, which results in plasma retention of solutes that are normally excreted by the kidneys . In the The increase in uricemia distribution quartiles was accompanied by increase in body adiposity markers . Altered WC was positively associated with urate concentrations; however, when fitted for other MS components, the effect was lost, thus showing that WC is indirectly associated with UA, since individuals with abdominal adiposity could present MS and/or alteration in its components, and these could be responsible for affecting UA. It is believed that TG is the main MS component influencing UA, as it was the only factor that remained significant after all adjustments.et al., (2003) [et al., (1999) [Other studies showed the relationship between WC or abdominal adiposity and UA increase ,45, repo, (2003)  and Frue, (1999) , where U, (1999) .2 was one of the main components associated with UA increase both in the fitted models and in the multiple regression analysis for males, and it also showed significant correlation with uricemia. BMI showed a positive relation with leptin concentrations [BMI above 25 kg/mtrations , which iAdditionally, individuals with high BMI may show insulin resistance, TG alteration and high blood pressure, and all these factors are related to UA increase . In our UA increase is observed in individuals with insulin resistance, probably because hyperinsulinemia would cause lower renal UA excretion . Besidesde novo synthesis of fatty acids [et al. (1998) report that the synthesis of fatty acids in the liver is related to the de novo synthesis of purine, thus accelerating UA production [In the present study, individuals with altered TG showed approximately 2.5 more chances of UA increase, regardless of other variables . Some studies show that high plasma triglyceride concentrations are related to hyperuricemia -55. One ty acids . Matsuuroduction .In the present study, UA concentrations were negatively determined by MM (kg) in males and females. The hypothesis would be the negative correlation existing between MM and inflammation , since aA recent study observed an inverse relation between MMI and UA in healthy individuals older than 40 years . The autIn-vitro studies showed that UA exerts a pro-inflammatory effect, thus stimulating the production of interleukin-1, interleukin-6 the tumor necrosis factor [A positive association was found between UA and CRP (inflammation), regardless of body composition (hyperadiposity and/or sarcopenia), gender, age and the presence of MS and its components, which means that there are a direct relation between these two factors. Another study showed a positive association between inflammation and UA, but no adjustment was performed to observe whether the effect would remain the same . In-vitrs factor .Individuals with higher CRP concentrations showed increased UA; however, UA is positively associated with total plasma antioxidant capacity, thus showing beneficial effects. On the other hand, deleterious relations of urate, such as the inverse association with adiponectin and the positive relation with E-selectin, were previously observed . Such asReduced HDL-c was one of the main factors responsible for UA increase (negative association) in females. This fact was observed by the backward stepwise multiple regression analysis. The negative correlation between HDL-c and AU was previously described , and it It is expected that individuals with high UA will have more chances of presenting MS . MS is aIn the present study, diet did not show direct influence on UA, but inadequate diet, conjointly with lack of physical activity, could alter body composition (higher adiposity), and such alteration would change UA. Additionally, an indirect relation of high carbohydrate intake was observed through the possible alterations in TG and/or glycemia.The intake of protein, meat and legumes, which could be related to increased purine intake, was not related to UA concentrations, thus agreeing with the literature ,14. SomeA significant weak and inverse correlation was observed (r = −0.11) between the intake of dairy products and UA, but when fitted for gender, BMI and VCT, the significance was lost. Although the present study did not observe such relation, other investigations reported an inverse association between dairy products and UA ,14,68, wBased on our data, lifestyle-modification conducts targeted at lean-mass maintenance and fat-mass reduction and concern about dietary composition are suggested in primary care for uricemia increase in non-uremic individuals.This was a cross-sectional study, and some cause/effect relationships cannot be possibly confirmed, but only whether or not an association between the studied factors exists. The dietary intake investigation on the individuals was performed only on one day, and a food recall for at least three weekdays would ideal, since there may be dietary variations that were not analyzed. Some dietary factors were not evaluated, such as the intake of alcohol, purine and caffeinated drinks, which are known for their interference with UA values ,68,70. IThe main factors associated with UA increase were altered BMI (overweight and obesity), muscle hypotrophy (MMI), higher levels of urea, triglycerides, and CRP. No dietary components were found among uricemia predictors.BMI: Body mass index; MMI: Muscle Mass Index; WC: Waist circumference; NCEP-ATPIII: National Cholesterol Education Program-Adult Treatment Painel III; MS: Metabolic Syndrome; γ–GT: γ-glutamyl transferase; CRP: C-reactive protein; TG: Triglycerides.The authors declare that they have no competing interests.EPO wrote the manuscript, FM corrected the manuscript, LVAS made the statistical analysis and corrected the manuscript. RCB read, corrected, and approved the final version of the manuscript. All authors read and approved.


Additionally, HTLV-1 carriers with dermatological skin lesions showed more frequently high proviral load as compared to asymptomatic carriers. The elevated proviral load in HTLV-1 patients with infectious skin lesions correlated significantly with TNF-α/IL-10 ratio, while the same significant correlation was found for the IL-12/IL-10 ratio and the high proviral load in HTLV-1-infected patients with autoimmune skin lesions. All in all, these results suggest a distinct and unique immunological profile in the peripheral blood of HTLV-1-infected patients with skin disorders, and the different nature of skin lesion observed in these patients may be an outcome of a distinct unbalance of the systemic inflammatory response upon HTLV-1 infection.In the present study, the frequency, the activation and the cytokine and chemokine profile of HTLV-1 carriers with or without dermatological lesions were thoroughly described and compared. The results indicated that HTLV-1-infected patients with dermatological lesions have distinct frequency and activation status when compared to asymptomatic carriers. Alterations in the CD4 In the present study, the immunological profiles of HTLV-1 carriers with or without dermatological lesions were thoroughly described and compared. The results indicated that HTLV-1-infected patients with dermatological lesions have distinct frequency and activation status than asymptomatic carriers. Alterations in cells and molecules that are important for immune cell function were observed in the HTLV-1-infected group with skin lesions. Additionally, HTLV-1 carriers with dermatological skin lesions have elevated frequency of high proviral load as compared to asymptomatic carriers, which indicates that the virus may be present in higher frequency in those patients. Patients with different skin lesions, autoimmune or infectious, also demonstrated differences in their immunological profile. All in all, these results suggest a distinct and unique immunological profile in the blood of HTLV-1-infected patients with skin disorders, and the different nature of skin lesion observed in these patients may be an outcome of a distinct unbalance of the systemic inflammatory response upon HTLV-1 infection. Human T-lymphotropic virus 1 (HTLV-1) was the first retrovirus discovered with human clinical importance + T cells in vitro and the viral particles are scarce in the peripheral blood of HTLV-1 carriers. In spite of that, HTLV-1 proviral DNA is detected particularly in CD4+ T cells of the host and the virus is able to induce strong activation and proliferation of many subsets of cells, which allow this retrovirus, together with other factors, to persist by clonal expansion of CD4+ T cells of the infected host + and CD8+ T cells according to Brito-Melo and collaborators (2007) Contrasting with HIV, HTLV-1 has poor infectivity of CD4There is still many unresolved questions on the context of the immunological status of HTLV-1 carriers who develop inflammatory symptoms, but there is no doubt that HAM/TSP is evidently a clinically and immunologically distinct entity that should be considered as unique among other inflammatory disorders associated with HTLV-1. The dermatological disorders associated with HTLV-1, however, have been poorly investigated, which poses a challenge on the management – prognosis and treatment - of patients with such disorders.Contrasting with the lack of scientific findings on the mechanisms that lead to dermatological disorders in HTLV-1 carriers, infectious and autoimmune dermatitis have been increasingly reported on those subjects, especially in endemic areas In face of the elevated prevalence of dermatological disorders among HTLV-1-infected individuals and the scarce knowledge of their immunological status, this study aimed at evaluating frequency, activation and the cytokine and chemokine profile of HTLV-1 carriers with or without dermatological diseases.Informed written consent for laboratory tests was obtained from each and all participants. The HEMOMINAS Foundation, the René Rachou Institute and Federal University of Minas Gerais (number 188/06) Ethics Committees approved the research. This study fulfilled recommendations 196/1996 from the Brazilian National Health Council for research involving humans.A full description of the patients included in this study is given in For FACS immunostaining, antibodies including: anti-human CD3-FITC, anti-human CD4-FITC, anti-human CD8-FITC, anti-CD14-FITC, anti-human CD16-PE, anti-human CD19-FITC, anti-human CD56-FITC, and anti-human HLA-DR-PE  were utilized. Whole blood staining of individual samples was performed according to manufacturer's instructions and adapted as described For acquisition and analysis, identification of the subsets was performed by the dual-color immunophenotyping method within the lymphocyte or monocyte scatter gate. All results were expressed as percentage of positive cells for the different subsets of cells analyzed in this study.To assess the levels of the cytokines – TNF-α, IL-12, IL-10, IL-6 and IL-1β – and chemokines – CCL2 (MCP-1), CCL5 (RANTES), CXCL8 (IL-8), CXCL9 (MIG), CXCL10 (IP-10) – in the sera from HTLV-1 carriers and controls, Cytometric Bead Array kits  were utilized according to manufacturer's protocol and adapted as described 4 and expressed as the number of HTLV-1 copies/104 cells.To quantify the HTLV-1 proviral load of the HTLV-1 seropositive individuals, peripheral blood was collected from the patients in tubes containing EDTA anticoagulant. DNA was isolated from peripheral blood by column extraction  and HTLV-1 proviral load was quantified by a real-time SYBR Green PCR method as previously described P values <0.05 were considered as statistically significant and indicated in the figures as letters or a line connecting the two different groups.The ANOVA one-way with Dunnet's post-test was utilized to compare the groups for all the immunological parameters evaluated. Pearson's and Spearman's correlation tests were utilized to compare cytokine levels with HTLV-1 proviral load of the patients. The Prism GraphPad Software version 5.0  was applied for the statistical analysis, and differences between groups with + T cells, and increased T/B cell ratio when compared to the HTLV-1-infected group with autoimmune skin lesions. The HTLV-1-infected group with autoimmune skin lesions showed increased levels of CD4+ HLA-DR+ T cells when compared to the HTLV-1-infected group without lesions.+CD16+/CD14+), pro-inflammatory monocytes (CD14+CD16+HLA-DR++/CD14+CD16+), NK cells (CD3−CD16+/−/CD56+/−/CD3−), NKT cells (CD3+CD16+/−/CD56+/−/CD3+) and regulatory T cells (CD4+CD25+High) were analyzed in the peripheral blood of HTLV-1 carriers with and without lesions as well as control , medium  or low proviral load . The frequency of patients with high, medium or low proviral load from each group demonstrated that HTLV-1 carriers without skin lesion contained the lowest percentage of patients with high-medium proviral load and the highest percentage of patients with low proviral load.The proviral load of peripheral blood mononuclear cells from HTLV-1-infected patients with or without skin lesions was evaluated. To evaluate whether the immunological features screened in the present study correlated with presence of the HTLV-1 provirus, the chemokines and cytokines levels were compared among patients of each group subdivided previously as high-medium and low proviral load carriers . Among aDermatological alterations are quite common in HTLV-1-carriers. Several reports have described dermatological diseases such as infective dermatitis as a prodromic manifestation of HAM/TSP and ATLL progression tax sequences However, the association between HTLV and Mycosis fungoides has remained controversial since few patients with mycosis fungoides are seropositive for antibodies to structural components of HTLV 1 and 2 virions and not all of the patients present evidence of + T cells and the T/B cell ratio are characteristic of HAM/TSP patients Brito-Melo (2002 and 2006) and Coelho-dos-Reis (2007) and collaborators proposed that, among the many altered subset of cells on HTLV-1 carriers from Brazil, the decreased frequency of B cells, the increased frequency of activated CD8In the present study, it was identified that patients with infectious dermatological lesions had similar frequency and activation status to HAM/TSP patients previously described, however, the chemokine profile seems to be divergent considering previous findings. Previous studies have demonstrated that levels of serum chemokines discriminate clinical HAM/TSP disease from HTLV-1 carrier state +CD16+/CD14+) were increased in the HTLV-1 group with skin lesions while the proinflammatory monocytes (CD14+CD16+HLA-DR++/CD14+CD16+) were decreased in both HTLV-1-infected groups with and without skin lesions. These results are in agreement with previous results that showed downregulation of CD14 exerted by HTLV-1 infection and impairment of differentiation of monocytes into macrophages or dendritic cells from HTLV-1-infected patients The monocytic populations seem to be altered in HTLV-1 patients with skin lesions. The macrophage-like monocytes , we merged the patients with high and medium proviral load to allow for a better understanding of the association of proviral load and skin lesion. In this analysis, the group with skin lesions presented higher frequency of patients with the higher-medium proviral load (14/27) as compared to the group without lesion (3/9).+ T cell frequency and cytokines such as IFN-γ and TNF-α was comprehensively described in patients with HAM/TSP. However, little is still known about this equilibrium in other HTLV-1-associated diseases. In agreement with previous findings on patients with myelopathy, the elevated proviral load in HTLV-1 patients with infectious skin lesions correlated significantly with TNF-α/IL-10 ratio, while IL-12/IL-10 ratio seems to be the ratio driving the unbalanced cytokine response in patients with autoimmune skin lesions. The reason that determines why the cytokine balance is different in the infectious versus autoimmune skin lesions still remains to be investigated. One hypothesis to explain this difference in cytokine balance could be the presence of different T cell subsets in the peripheral blood of patients with skin lesions. While the HTLV-1-infected group with infectious dermatological lesions presented increased frequency of CD8+ T cells, the HTLV-1-infected group with autoimmune skin lesions showed increased levels of CD4+ HLA-DR+ T cells. These different T cell populations could provide a microenvironment that modulates the production of TNF-α and especially IL-12 from macrophage-like monocytes, which are increased in patients with skin lesions Click here for additional data file.


Although the importance of communication is recognized in several disciplines, it is rarely studied in the context of online social interactions and joint actions. During online joint actions, language and gesture are often insufficient and humans typically use non-verbal, sensorimotor forms of communication to send coordination signals. For example, when playing volleyball, an athlete can exaggerate her movements to signal her intentions to her teammates  or to feint an adversary. Similarly, a person who is transporting a table together with a co-actor can push the table in a certain direction to signal where and when he intends to place it. Other examples of “signaling” are over-articulating in noisy environments and over-emphasizing vowels in child-directed speech. In all these examples, humans intentionally modify their action kinematics to make their goals easier to disambiguate. At the moment no formal theory exists of these forms of sensorimotor communication and signaling. We present one such theory that describes signaling as a combination of a pragmatic and a communicative action, and explains how it simplifies coordination in online social interactions. We cast signaling within a “joint action optimization” framework in which co-actors optimize the success of their interaction and joint goals rather than only their part of the joint action. The decision of whether and how much to signal requires solving a trade-off between the costs of modifying one’s behavior and the benefits in terms of interaction success. Signaling is thus an intentional strategy that supports social interactions; it acts in concert with automatic mechanisms of resonance, prediction, and imitation, especially when the context makes actions and intentions ambiguous and difficult to read. Our theory suggests that communication dynamics should be studied within theories of coordination and interaction rather than only in terms of the maximization of information transmission. The study of human communication is central in several disciplines, including linguistics, cognitive science, neuroscience, anthropology, biology, and philosophy human sensorimotor communication during online social interactions and joint actions, such as for instance when two persons play volleyball or jointly lift a table. Unlike conversation, these joint actions are not exclusively communicative but have non-communicative, pragmatic goals (winning a match or transporting the table somewhere). Still, as recognized by early scholars Less attention has been devoted to In online sensorimotor interactions and joint actions, communication is not only linguistic. Several studies have focused on other forms of communication, which include deictics, gesturing, and facial expressions, see signaling something to a co-actor ? Evidence accumulates that distinguishing features can be found at the kinematic level. For example, it has been reported that in social contexts actions have subtle still significant kinematic peculiarities: the deceleration phase was slower when a “giving” action was directed to another individual that in an individualistic set-up (placing an object) How can we distinguish when a given action  is used for its pragmatic goal  or for Lombard effect). Studies of sign language reveal dissimilation effects, too: performers chose movement parameters that make two successive actions more easily discriminable for the perceiver Sensorimotor strategies of communication can also work synergistically with other forms of communication . It has been consistently reported that child-directed speech (a “motherese”) has certain characteristics that facilitate recognition and perceptual processing, such as for instance an exaggeration of the vowels Despite these empirical demonstrations, the nature and functioning of human sensorimotor communication is largely unknown. There is currently no normative framework or analytic description of human signaling and sensorimotor communication that provides them with a theoretical ground. Besides its importance per se, the study of sensorimotor communication provides an excellent opportunity to understand the adaptive and evolutionary value of communication in terms of coordination and interaction success. Indeed, joint action scenarios are interesting because they are not essentially communicative but have first and foremost pragmatic goals. This makes it possible to study the trade-offs between the costs and benefits of communication in terms of interaction success, not (only) of communication success, as it is instead common in theories focusing on maximization of information transmission.In this article we present a formal theory of signaling in online sensorimotor interactions. Signaling refers to the intentional modification of one’s own behavior to convey information to another person, typically a co-actor. In addition to a pragmatic goal , the signaler generally has the communicative goal of changing the co-actor’s belief . In some cases, but not always, the signaler wants also the co-actor to be aware that she is intentionally communicating or cooperating. Note that in competitive scenarios the signal can also be misleading  rather than their individual ones , and take into consideration the joint outcome  and when necessary also the parameters of the co-actor  rather than only their own. Signaling is part of this We formalize signaling in terms of parametrizable deviations from the action’s optimal trajectory so that the signaling action retains its pragmatic goal  but the changes in the kinematic parameters are informative of the performer’s action choice. To choose whether or not to signal, and how much to signal, subjects perform a cost-benefit analysis . As it requires a cost-benefit analysis, we consider signaling as an intentional form of sensorimotor communication, not a byproduct of interaction dynamics.dissimilation (or differentiation) of one’s own action parameters  from those of the alternative actions that are most likely given the context and the perceiver’s prior information. Fourth, our parametrization permits to select different “degrees” of signaling actions, and to consider their costs  in addition to their benefits .Our theory makes four assumptions. First, the computational objective of signaling is permitting the co-actor to better discriminate the signaler’s action goal  against the alternatives  the action recognition process of a perceiver agent. Consider the case of a volleyball player who can execute one of two possible action alternatives: passing the ball to the left or right. A teammate sees the scene but is uncertain relative to the performer’s action: action 1  or action 2 . The teammate can estimate the most likely action of the performer based on multiple sources of information, such as the performer’s arm trajectory, body posture, hand movements, gaze direction, and various kinds of prior information (e.g. the performer’s preference and skills). The essence of our signaling theory is that the performer can Let’s describe more formally the computational problems that performer and perceiver must solve. Each goal-directed action can be performed in a number of different ways . An authoritative view in computational motor control is that each particular instantiation of an action can be associated to an internal model During an interaction, the perceiver’s goal is to infer which model signaling.The performer’s goal, on the other hand, is to facilitate such an inferential process. Here we argue that this result can be achieved by actively modifying the parameters of the currently executed model, and thus the resulting trajectory dissimilation is defined as the amplification of the differences between the selected trajectory and the alternative one(s). At the same time, however, the action choice has to be a valid one to preserve its pragmatic effect  and make the recognition by the perceiver possible. Thus, samples from the signaling distribution for a given model Signaling can be defined as the process of altering one’s own behavior to facilitate its recognition by other persons. In our framework, a signaling agent tries to “dissimilate” her trajectory from the alternative ones that are possible (or likely) given the context rejection sampling technique accept the sample In line with the requirements above, samples from the signaling distribution can be obtained via a modified signaling distribution:In the case of continuous distributions Intuitively, the signaling distribution for the model The minimization of Eq. 4 considers jointly three factors. The KL term considers the cost of signaling, where cost can be associated to biomechanical factors, effort, and other forms of costs . The As an example of signaling distribution, consider a system governed by two independent Gaussian processes corresponding to models For the sake of simplicity we assume that the contribution of each internal model to the state can be expressed by a Gaussian distribution with mean However, the theory is independent from the particular density function chosen.To test our model, we performed three experiments. The first experiment uses a synthetic dataset to tests if, when the performer uses the signaling strategy elucidated earlier, the perceiver can better discriminate the right action from an alternative one. The second experiment consists in using our proposed approach to model human data on a joint action task that requires signaling. The third experiment is similar to the first experiment but considers the case of three actions to be disambiguated.To test the efficacy of the proposed signaling model in terms of improved action recognition, we designed an experiment using a synthetic dataset with two trajectories corresponding to two goal-directed actions: signaling distribution is used instead. By comparing signaling distribution. The synthetic perceiver agent is modeled using a Bayesian action recognition method that integrates over time (noisy) trajectory estimates, see We argued that the effect of signaling and dissimilation is minimizing the probability of misclassification of a perceiver agent. To assess this hypothesis, we compared the performance of a (synthetic) perceiver agent asked to recognize a performed action when its trajectory is sampled from (i) the original distribution and (ii) a Leader, knows the task to be jointly performed . Results show that in this case the Leader modifies her trajectory to signal her co-actor, called the Follower, the task to be performed. Specifically, if the Leader is required to do an up-reaching, she significantly raises the trajectory; if she is required to do a down-reaching, she significantly lowers the trajectory. The objective of experiment 2 is is assessing if our method can be used to model human signaling data. A recent experiment reaching up and reaching down. This was done by using the baseline human data in a supervised learning scheme, adopting the Echo State Gaussian Process algorithm We used the proposed signaling method to model the human data reported in signaling distributions using different values of profile in Then, we calculated various Up to now we have considered the case of two actions These results show that signaling strategies are flexible and tailored to the action alternatives. The success of a signaling action depends on the action alternatives that are considered. Let’s imagine that a perceiver action is uncertain between A1 and A3. If the performer uses a signaling action that disambiguates between A1 and A2 rather than A1 and A3, he would stop signaling too early and this would make the perceiver’s task harder. We hypothesize that the performer modifies its signaling strategies using an estimation of the perceiver’s belief of the possible alternatives and the associated uncertainty. This novel empirical prediction of our model remains to be studied empirically. However, ongoing joint action experiments with several action alternatives show that the leader’s movement amplitude contains information about the target location (which is unknown to the co-actor) and the amplitude increases when the target-to-target distance increases .joint action optimizationAlthough the importance of communication is recognized in several disciplines, it is rarely studied in the context of on-line social interactions and joint actions. We offered a normative framework in which a performer agent can decide to pay a cost in order to send coordination signals and help the co-actor’s recognition process. In this framework, signaling is not conceptualized as an “altruistic” action  but as part of a strategy for Recent research in social neuroscience has revealed that understanding the intentions of co-actors and predicting their next actions are fundamental for successful joint actions and social interactions, cooperative and competitive. In joint task such as building something together or running a dialogue, action understanding helps planning complementary actions and contributes to the success of the joint goal. Furthermore, predictive mechanisms help the real-time coordination of one’s own and the co-actor’s actions intentional strategies to enhance coordination. In keeping with this view, human experiments show that signaling is versatile and takes into account the communicative implications of the executed actions, its ambiguity and its costs, as well as the uncertainty of the co-actor However, action understanding and prediction are hard and often under-constrained computational problems, and it is still unclear how humans  solve them in real time while at the same time planning their complementary or competitive actions plan rather than only her next action We argue that signaling strategies help solving coordination problems and joint actions by making actions easier to predict and disambiguate. In the long run, signaling actions can also support common ground formation by disambiguating an actor’s intended An open objective for future research is investigating empirically the multifarious methods humans use for sensorimotor communication and signaling, which are not fixed but depend on task demands and goals When communication is studied in purely communicative scenarios  it is tempting to argue that it maximizes information measures such as the correct reproduction of a message, as indeed assumed by prominent frameworks such as Communication Theory The theory we have presented does not require that the perceiver recognizes the communicative nature of the performer’s action, i.e., that the performer intended to communicate. Still, several researchers argue that recognizing the communicative intention is be a fundamental element of pedagogical contexts The current model has some limitations that should be addressed by future research. First, we adopted a simplified action execution model that does not consider action biomechanics, muscle activation, and detailed planning mechanisms. Furthermore, for the sake of simplicity we assumed that the costs of signaling  are proportional to the amount of the deviation from the “original” trajectory. These limitations can be overcome by introducing in our formulation more detailed action models based on optimal control theory decision theory and reward-directed motor control game theory, which is an established framework for studying coordination problems. in this vein, a few recent studies linked sensorimotor or cognitive processes to equivalent game-theoretic concepts, but did not consider communication joint action optimization resonates with game-theoretic concepts of team reasoningAnother promising direction for future research is linking more directly the proposed approach to established frameworks in computational motor control and social interaction. The proposed theory can be linked to existing models of Bayesian


These results suggest that glucosylceramide plays an important role in anti-inflammatory and memory enhancement, and it could be a potential new therapeutic agent for the treatment of neurodegenerative diseases such as Alzheimer's disease.The function and the role of glucosylceramide have not been well studied in the central nervous system. This study was aimed to investigate the possible roles of glucosylceramide in memory function in aged mice. Glucosylceramide  showed memory enhancing activity after 3-month treatment in the aged mice  through Y-maze, novel objective test, and Morris water maze test. Long-term treatment of glucosylceramide decreased the expression of iNOS and COX-2 in the brain of aged mice. The LPS-induced mRNA level of iNOS, COX-2, IL-1 Carbon chemical shifts are reported in ppm relative to TMS with the respective solvent resonance as the internal standard (CDCl3). Infrared (IR) spectra were recorded on a JASCO FT/IR-430 spectrometer. Data are reported in wave numbers (cm−1). Melting Points were determined on a BIBBY Stuart Scientific Melting Point Apprataus SMP3. Mass spectra were recorded with a Waters Micromass ZQ LC-Mass system and high resolution mass spectra (HRMS) were measured with a Bruker BioApex FTMS system by direct injection using an electrospray interface (ESI).Glucosylceramide was readily characterized through physicochemical instruments after purification by flash column chromatography. Flash column chromatography was performed with Merck silica gel 60 (230–400 mesh) for purification. −1; 1H NMR  δ 5.60–5.52 , 5.42–5.28 , 5.01–4.87 , 4.55–4.48 , 4.13–4.09 , 4.04–3.99 , 3.85–3.78 , 3.54–3.28 , 2.50 , 2.05–1.90 , 1.54–1.45 , 1.33–1.19 , 0.88–0.81 ; 13C NMR  δ 173.75, 132.31, 130.23, 129.73, 127.76, 76.89, 76.53, 73.11, 71.59, 70.01, 33.91, 33.22, 32.54, 31.99, 31.29, 30.89, 29.03, 28.71, 28.58, 26.59, 25.21, 24.43, 22.09, 21.96, 14.11; HRMS calcd. for C40H76NO9: 714.5520 [M+H]+, found: 714.5531.Glucosylceramide: mp: 195-196°C; IR  3353 (O–H), 2920 (C–H), 2851, 1726 (C=O), 1620 (C=C), 1530, 1466, 1406, 1378, 1264, 1182, 1168, 1040 (C–O), 721 cmSpontaneous spatial recognition in the Y-maze was used as a hippocampus-dependent test. The Y-maze is a three-arm horizontal maze  in which the arms were symmetrically disposed at 120° angles from each other. The maze floor and walls were constructed from dark opaque polyvinyl plastic. Mice were initially placed within one arm, and the sequence  and number of arm entries were recorded manually for each mouse over 8-minute period. An actual alternation was defined as entries into all three arms on consecutive choices . Maze arms were thoroughly cleaned between tasks to remove residual odors. One hour after the last administration of glucosylceramide or saline alone, memory impairment was tested. Mice were gently placed in the maze. The percentage of alternations was defined according to the following equation: %alternation = [/] × 100. The number of arm entries served as an indicator of locomotor activity.T) an exploration index was calculated: [(TNovel − TFamiliar)/(TNovel + TFamiliar)] × 100. This measure is considered as an index of recognition memory and takes into account individual differences in the total amount of object exploration time.This test was used to measure for objective recognition. The arena was a cage bottom (30 × 40 × 20 cm) with black walls. The objects were the same size but differing in shape, color, and surface texture. On day 1, each mouse was habituated individually to the test box for 8 min sessions in which the animals were allowed to freely explore the open field box. After 6 h, two identical objects were placed in each corner and the mouse was allowed 8 min to explore the objects. Pairs of each object were used for an equal number of times. On day 2 one object was replaced with a novel object in a counter-balanced fashion with respect to object, side, and genotype. The mouse was allowed 8 min to explore the familiar and novel object while it was video-recorded. Exploration was defined as sniffing the object or having one or both forepaws touching the object. From the absolute exploration duration  and centrifuged at 13500 rpm for 15 min twice at 4°C. Samples were assayed for protein concentration using protein assay kit . Proteins were separated by SDS-polyacrylamide gel electrophoresis and transferred to a polyvinylidene difluoride membrane. The membrane was blocked with 5% nonfat dry milk in Tris-buffered saline/Tween 20 solution. The blots were incubated with the iNOS and COX-2 . GAPDH  was performed as an internal control. After washing with Tris-buffered saline/Tween 20, horseradish peroxidase-conjugated secondary antibodies  were applied, and the blots were developed using the enhanced chemiluminescence detection kit .The brain tissue was homogenized with homogenization buffer  and TNF-α ; PCR products were separated by 1% agarose gel electrophoresis and visualized by ethidium bromide staining.Mice were stimulated with LPS in the absence or presence of glucoceramide for 18 h. Total RNA was isolated from hippocampus of mice using TRIzol (Invitrogen) according to the manufacturer's instructions. For cDNA synthesis, 2 ug of total RNA was reverse-transcribed using the SuperScript First-Strand Synthesis System (Invitrogen). cDNA was amplified by polymerase chain reaction (PCR) using primers for iNOS , COX-2 , IL-1P < 0.05 were considered as statistically significant to analyze the difference.All values were expressed as mean ± standard error (SE). The results were subjected to an analysis of the variance (one-way ANOVA) using the Newman-Keuls multiple comparison test. Differences with *F  = 8.388, P = 0.0021, one-way ANOVA]. Glucosylceramide resists the working memory retention. There was significant effect on the percentage of alternation  were tested in the Y-maze test. Mice were administrated for 3 months with glucosylceramide (50 mg/kg) which mixed in the chow. Aged mice showed the impairment in working memory function since aged mice spent less spontaneous alteration than young mice in Y-maze test. Glucosylceramide enhanced spatial memory since glucosylceramide-treated mice spent higher spontaneous alteration in the novel arm than that of saline group . The amount of time spent with the novel object compared with the total time spent exploring both object represents an index of long-term memory. Mice were administrated with glucosylceramide for 3 months (50 mg/kg). Treatment of glucosylceramide enhances memory function on the performance of a novel object recognition task . As expeP < 0.05). On the day following the final day of training trial sessions, swimming times within the platform quadrant for the glucosylceramide group were significantly lower than those of the vehicle-treated young group (P < 0.05)  = 17.69, P = 0.0001, one-way ANOVA]. However, no significant differences in swimming speeds were observed between the groups. In this experiment, aged mice increased the escape latency time at the training sessions; however, glucosylceramide treatment shortened this escape latency time on day 4. At the probe trial session, glucosylceramide treatment increased the swimming time within the target quadrant. The decrease in escape latency from day to day in the first trial represents long-term memory or reference memory, while that from first trial of sessions to second trial of sessions represents short-term memory or working memory [The effects of glucosylceramide on learning and spatial memory were evaluated by the Morris water maze test. Mice were administrated with glucosylceramide (50 mg/kg) for 3 months. The aged group exhibited longer escape latencies throughout the training days than that of the young group. Glucosylceramide group significantly shortened the escape latencies which was prolonged by aged group ( < 0.05) . Moreoveg memory . The timg memory . These rGlucosylceramide exerted an anti-inflammatory effect on age-related responses accompanied by the expression of iNOS and COX-2. Aged mice were administrated with glucosylceramide for 3 months. The protein expression levels of the iNOS and COX-2 in hippocampus of brain were reduced by administration of glucosylceramide . These rβ, and TNF-α were reduced by treatment with glucosylceramide  , 22. ButIn the present study, it was shown that long-term administration of glucosylceramide ameliorated the memory decline and anti-inflammatory activity in aged mice. The memory enhancing effect of glucosylceramide was confirmed in the Y-maze test, novel objective test, and Morris water maze task in different behavioral observation. In the Morris water maze test, glucosylceramide increased the swimming time within the target quadrant. The decrease in escape latency from day to day in the first trial represents long-term memory or reference memory . The timMice fed with the control diet displayed decreasing spatial memory performances with age. This was confirmed to increase markers of inflammation such as iNOS and COX-2 as well as mRNA levels in the hippocampus and inhibit hippocampal-dependent working memory. However, administration of glucosylceramide decreased the age-associated expression of proinflammatory mediators such as iNOS and COX-2 in the aged mice brain and performance in the hippocampal-dependent working memory task was restored. Thus, age-related conditions that are exacerbated by inflammation, including cognitive aging and neurodegenerative diseases, may be inhibited by dietary intake of glucosylceramide. Also pretreatment of glucosylceramide suppressed the LPS-induced proinflammatory responses; iNOS, COX-2, and cytokines in mice.trans-double bond between C-4 and C-5. There is an interesting report that phytoceramide activates peroxisome proliferator-activated receptors (PPARs), whereas ceramide and dihydroceramide do not change the PPAR activity [Ceramide has been known as a sphingolipid with potent proinflammatory and proapoptotic properties –43 and iactivity .The structural feature of glucosylceramide is represented as attached glucose on terminal hydroxyl group while the normal ceramide consists of sphingoid base linked to a fatty acid with an amide bond. In this point the glucosylceramide would be hydrolyzed by glycosidases in vivo to generate ceramide. Even though general ceramide is formed as potential component in biosynthesis of many complex sphingolipids, the biological activities of ceramide showed relatively variable activity depending on the cell types. Glucosylceramide had better solubility than that of ceramide and showed better bioavailability. Interestingly, phytoceramide and glucosylceramide (after hydrolysis) have three –OH groups but ceramide has two hydroxyl groups. Collectively, glucosylceramide plays an important role in anti-inflammatory and memory enhancement, and it could be a potential new therapeutic agent for the treatment of dementia.


While human T cell leukemia virus type 1 (HTLV-1) is an etiological agent of adult T-cell leukemia (ATL), its close relative HTLV-2 is not associated with any leukemia. HTLV-1 and HTLV-2 encode Tax1 and Tax2 proteins, respectively, which are essential for immortalization of T-cells by the respective viruses, thereby persistent infection. Tax1 and Tax2 have more than 75% amino acid similarities, but we show here that Tax1 and Tax2 have multiple distinct activities. Tax1 induced IL-2-independent growth of a T-cell line CTLL-2 more efficiently than Tax2. By contrast, Tax2 immortalized human T-cells in the presence of IL-2 more efficiently than Tax1. These results suggest that HTLV-1 and HTLV-2 have distinct IL-2 requirements to immortalize T-cells. We also found that Tax1 altered cellular oxidative stress response. Arsenite, a pro-oxidant, induced stress granule (SG) formation, which functions as a protective role against oxidant-induced cell damages, but the formation was inhibited by Tax1. We will discuss these findings in terms of the HTLV-1-specific pathogenesis.


A primary route of transmission of human T-lymphotropic virus 1 (HTLV-1) is from mother-to-child via breast milk, but knowledge of the early immunologic events in orally acquired HTLV-1 infection is limited. Herein, we characterized normal rabbit gut-associated lymphoid tissues (GALT) and performed studies to develop an oral model of HTLV-1 infection. Mononuclear leukocytes were immunophenotyped from key GALT inductive and effector sites using flow cytometry and immunohistochemistry. Our data indicate that unexposed rabbits GALT have a predominant CD4+ lymphocyte population similar to humans. To establish a HTLV-1 oral model 12 week old female New Zealand White rabbits were orally or intravenously inoculated with CD3+CD4+CD25+ rabbit lymphocyte cell line immortalized with the HTLV-1 molecular clone ACH (R-49 cells) or control Jurkat T-cells orally. The rabbits were monitored for hematologic and virologic parameters prior to serial sacrifice. Collectively, 66 to 100% of HTLV-1 orally exposed rabbits became persistently infected. HTLV-1orally exposed and infected rabbits during early time points (1-4 weeks post exposure) had delayed and often less intense anti-HTLV-1 antibody response, variable leukocytosis, and a delayed p19 matrix antigen production and proviral DNA amounts in peripheral blood leukocytes compared to IV exposed rabbits. Interestingly, by 8 weeks post exposure orally exposed rabbits had established similar systemic spread of the virus compared to IV exposed rabbits. This oral model of HTLV-1 transmission in rabbits creates to opportunity to test the role of the mucosal microenvironment during the early stages of orally-acquired HTLV-1 in gut-associated lymphoid tissue.


In Reply: We understand that the Croatian Medical Journal has received a Letter to the Editor with comments on our recently published study . We agree, and we did exercise a considerable caution. First, considering that multivariate models in different studies included different independent variables , we concluded that for a more precise/accurate estimate of the strength of the independent UA-acute myocardial infarction (AMI)-outcome association further studies were needed. We also concluded that further studies were needed in different settings, ie, STEMI, NSTEMI, mode of treatment etc. Second, we pointed-out that, particularly regarding the long-term outcomes , on-admission serum UA might not be of such an interest as long-term development of the UA values during the post-AMI period. Unfortunately, no single study so far has addressed serum UA as a time-varying factor (covariate). On the other hand, we believe that the fact that 9 separate studies  all found an independent association of UA and adverse AMI outcomes – speaks for itself. Indeed, 9 studies with around 8000 patients might be an insufficient number for a definite, robust generalization about the UA-AMI outcome relationship , but the results of the individual studies as well as of the pooled analysis are more than indicative.At the beginning of the second paragraph, Zhou and colleagues refer to 3 or 4 studies referring to the association between serum UA and coronary heart disease (CHD) and conclude that literature data are inconsistent regarding the question whether serum UA “predicts CHD.” In this respect, we would like to point out the following: a) our aim WAS NOT to assess whether serum UA was a predictor of occurrence of CHD , since this issue had been thoroughly addressed, rather, we addressed another question: do on-admission (within 48 hours since the symptom onset) serum UA levels predict the outcome in patients affected by AMI? b) As for the relationship between serum UA and CHD , we draw the attention to the meta-analysis published in 2010  properties of UA. This is a well known fact and is addressed also in our work. Still, the fact remains that PROLONGED HIGH(ER) UA levels clearly are adversely related to many different cardiovascular diseases (regardless of the possible mechanistic explanation or lack of it). Further, Zhou and colleagues refer to one study that investigated the relationship between serum UA and the outcomes in acute ischemic stroke (IS) patients. In this respect, we would like to point out: a) acute IS and acute MI, although with many common underlying features are two DIFFERENT diseases ; we addressed only MI and made no implications regarding IS; b) there are numerous studies investigating UA and IS. One meta-analysis of 16 studies with around 250 000 subjects found UA to be independently associated with a higher incidence and higher IS-related mortality . A numbeFinally, Zhou and colleagues warn about the “…limitations of meta-analysis…” due “...to non-random choice of individual studies”. We do not understand this comment at all: systematic review and meta-analysis IS ABOUT inclusion of ALL available studies (which we accomplished through a very thorough literature search) and their critical evaluation and, eventually (if feasible and justified), pooled analysis with a goal of obtaining the most realistic estimate (of an effect).To conclude, the relationship between serum UA levels and various aspects of cardio- and cerebrovascular diseases has been extensively investigated. We addressed only one simple question: Do serum UA levels taken on-admission in patients affected by acute myocardial infarction predict the outcome? The available data and the critical assessment and pooled analysis that we performed strongly suggest that serum UA should be considered as an independent predictor regarding the short-term, medium-term, and long-term outcomes in these patients.


The present review discusses intracellular signaling moieties specific to membrane lipid rafts (MLRs) and the scaffolding proteins caveolin and introduces current data promoting their potential role in the treatment of pathologies of the heart and brain. MLRs are discreet microdomains of the plasma membrane enriched in gylcosphingolipids and cholesterol that concentrate and localize signaling molecules. Caveolin proteins are necessary for the formation of MLRs, and are responsible for coordinating signaling events by scaffolding and enriching numerous signaling moieties in close proximity. Specifically in the heart and brain, caveolins are necessary for the cytoprotective phenomenon termed ischemic and anesthetic preconditioning. Targeted overexpression of caveolin in the heart and brain leads to induction of multiple pro-survival and pro-growth signaling pathways; thus, caveolins represent a potential novel therapeutic target for cardiac and neurological pathologies. Cardiac  and neuronal  diseases together represent a majority of mortalities Hoyert,  and sourMembrane lipid rafts (MLRs) are discreet microdomains of the cell membrane that concentrate and localize signaling molecules by providing a stable platform for protein anchoring. Stability of protein anchoring is afforded by the lipid composition of MLRs, which are rich in sphingomyelin, glycosphingolipids, and cholesterol. By promoting a favorable binding environment for cell signaling receptors and their downstream effectors, MLRs promote a variety of physiological functions such as cell surface signaling  is involved in the control of many important functions, such as angiogenesis, vasorelaxation, and permeability. Mice deficient in Cav-1 display increased nitric oxide (NO) production from increased eNOS activity  can be activated by GPCRs or tyrosine kinase receptors, and is intimately involved in cell growth and survival through activation of the anti-apoptotic Akt pathway. In cultured skeletal myocytes, disruption of MLR's impairs cell survival via inhibition of PI3K/Akt  Cav-3 overexpression system that caused a greater than twofold increase in Cav-3 specifically in cardiac myocytes and subjected the mice to transverse aortic constriction (TAC) for 1 month, to induce cardiac hypertrophy. Mice with cardiac-specific overexpression of Cav-3 exhibited augmentation of natriuretic peptide expression and nuclear Akt phosphorylation, resulting in reduced cardiac hypertrophy following TAC, with improved cardiac function and increased survival suggesting a potential therapeutic role for Cav-3 in heart failure , resulting in Ca2+ influx into the cytosol and subsequent contraction. However, current evidence suggest a more complicated system of basal oscillating “sparks” and “puffs” of cytosolic Ca2+ mediated by ligand-gated receptors , leading to destabilization of membrane polarization  Cav-3 overexpression demonstrated increased myocardial caveolar formation, augmentation of pro-survival kinases, and exhibited innate cardioprotection with improved cardiac function at a level similar to that achieved through IPC  receptor, and the glutamate receptor, which share the common final pathway of phosphorylation of the cAMP response element binding protein (CREB), initiating transcription of pro-growth genes Figure . Cav-1 aCerebral ischemia decreases the integrity of cell membranes and thus may disrupt caveolin-associated signaling complexes. A neuroprotective role for Cav-1 was first demonstrated in Cav-1 KO mice, which had a larger cerebral infarct size in response to ischemia  and neurodegeneration is limited by a reduction in pro-survival and pro-growth signaling  has great therapeutic/translational potential, the use of small molecules to induce cell specific enhancement in mRNA translation will allow us to achieve alterations in protein expression-independent of viral vectors. An example is the use of complementary inhibitors of miRs, termed “antagomirs.” In regard to caveolin-specific miR inhibition, one group showed that the use of antagomirs to block Cav-1 associated miRs (miRs 103/107) and subsequently enhance Cav-1 protein expression reversed a type II diabetic phenotype  are small non-coding RNAs that serve to regulate post-transcriptional gene expression by binding to mRNA and repressing translation or degrading mRNA entirely. They are highly abundant in the brain and have been known to localize to subcellular regions such as dendritic spines and can affect spine structure/morphology, function, and synaptic plasticity (Gao et al., 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.