Human T-lymphotropic virus type 1 (HTLV-1) is the causative agent of adult T cell leukemia (ATL) and other HTLV-1 associated neurological disorders. Unlike most retroviruses, cell-free HTLV-1 virions are poorly infectious and do not stably infect its primary CD4+ T lymphocyte target. However, HTLV-1 efficiently infects cells of the myeloid lineage, leading to productive infection of myeloid cells. Here, we investigate the mechanisms underlying monocyte infection by HTLV-1 and demonstrate that HTLV-1 infection induced apoptosis of monocytes in a SAMHD1-dependent manner. SAMHD1, a deoxynucleoside triphosphate triphosphohydrolase, functions as a restriction factor that limits HIV-1 replication by reducing the availability of deoxynucleosidetriphosphates (dNTPs) required for reverse transcription. RNAi-mediated silencing of SAMHD1 inhibited monocyte apoptosis, while addition of exogenous dNTPs, or pre-treatment with azidothymidine (AZT) to block reverse transcription also inhibited apoptosis. To investigate a role for reverse transcription intermediates (RTI) in triggering apoptosis, a biotinylated 90 nucleotide RTI from the U5 region of HTLV-1 was introduced into monocytes; strikingly, the biotinylated RTI induced apoptosis and bound to the DNA sensor STING - which mediates the antiviral response via IRF3 activation. We further demonstrated that STING-mediated apoptosis in infected monocytes required the generation of a pro-apoptotic complex between IRF3 and the Bcl-2 protein Bax. These studies provide a mechanistic explanation for HTLV-1 abortive infection of monocytes and report a link between SAMHD1 restriction of reverse transcription, sensing of retroviral reverse transcription intermediates by STING, and the initiation of IRF3-Bax driven apoptosis.


Infection with HTLV-1 is associated with bronchiectasis in Indigenous Australians. Previous studies have not determined HTLV-1 proviral loads (pvl), which predict risk of other HTLV-1 associated inflammatory diseases, such as HTLV-1 associated myelopathy. Thirty-six Indigenous adults admitted to Alice Springs Hospital, June 2008 to December 2009, with radiologically confirmed bronchiectasis, but no other HTLV-1 related disease, were prospectively recruited and matched by age and sex to 36 controls of the same ethno-geographic origin. Case notes were reviewed from date of birth or first admission to date of recruitment. HTLV-1 subtype C pvl were determined at the Pasteur Institute, Paris. HTLV-1 infection was more common among cases  than their controls (p<0.018). Two cases were admitted in childhood with probable infective dermatitis and two with recurrent strongyloidiasis unresponsive to treatment with thiabendazole. In adulthood, cases  were more likely than controls  to have positive or borderline strongyloides serology (p<0.001). The mean HTLV-1 pvl (±SEM) was significantly higher for cases (0.83±0.21% PBMC) than controls (0.12±0.05% PBMC)(p=0.016) and was nearly17 fold-higher compared to controls in which bronchiectasis was excluded by HRCT . Twelve cases (33.3%) and 5 controls (13.9%) died during 3 years of follow-up (p<0.052). Cases died at a younger age (p=0.15). In both groups HTLV-1 infected patients were more likely to die: cases, 11/12 (92%); controls, 4/5 (80%). HTLV-1-associated bronchiectasis is associated with higher HTLV-1 pvl suggesting that this condition results from an HTLV-1 driven inflammatory process.


Background. The human T-cell lymphotropic virus types 1 and 2 (HTLV 1/2) are retroviruses associated with different pathologies. HTLV-1 causes adult T-cell leukemia/lymphoma (ATL) and HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP); HTLV-2 is not clearly associated with a known clinical disease. Both viruses may be transmitted by whole blood transfusion, from mother to child predominantly through breastfeeding, and by sexual contact. Presently, none of the regional blood banks in Uganda perform routine pretransfusion screening for HTLV. The aim of this study was to determine the prevalence of anti-human T-cell lymphotropic virus types 1/2 (HTLV-1/2) antibodies among blood donors at Mbarara Regional Blood Bank in South Western Uganda. A cross-sectional study was conducted between June 2014 and September 2014. Methodology. Consecutive blood samples of 368 blood donors were screened for anti-HTLV-1/2 antibodies using an enzyme linked immunosorbent assay (ELISA). Samples reactive on a first HTLV-1/2 ELISA were further retested in duplicate using the same ELISA. Of the three hundred and sixty-eight blood donors (229 (62.2%) males and 139 (37.8%) females), only two male donors aged 20 and 21 years were HTLV-1/2 seropositive, representing a prevalence of 0.54%. Conclusion. HTLV-1/2 prevalence is low among blood donors at Mbarara Regional Blood Bank. Studies among other categories of people at risk for HTLV 1/2 infection should be carried out. Almost 10–20 million people in the world are thought to be infected by human deltaretroviruses, namely, human T-cell lymphotropic virus (HTLV) types 1 and 2 [Human T-cell lymphotropic virus type 1 (HTLV-1) is a known aetiological agent of adult T-cell leukemia/lymphoma (ATL) and HTLV-1-associated myelopathy (HAM)/tropical spastic paraparesis (TSP) ; howeverThough it has a worldwide distribution, HTLV-1 is endemic only in South Western Japan, the Caribbean Basin, West and Central Africa, some areas of intertropical Africa (such as South Gabon) and of the middle East (such as the Mashhad region in Northeastern Iran), rare isolated clusters in Australo-Melanesia, and foci in South America . HTLV-2 Serosurveys worldwide have documented that HTLV-1 and HTLV-2 are transmitted by contaminated blood transfusion , breast HTLV-1 and HTLV-2 virus transmission through blood transfusion is considered the most efficient route and the seroconversion of the recipients is around two months .Seroconversion has been observed in 44%–63% of cases after receiving blood contaminated with HTLV-1 infected cells . EffortsScreening has been shown to reduce risk of transfusion related transmission in USA . It was The study was carried out at Mbarara Regional Blood Bank in Mbarara Municipality, South Western Uganda, from June 2014 to September 2014.A total of 368 blood samples from the blood donors selected by consecutive nonprobability sampling method were included in this study. Blood donors were selected by the regional blood bank if they fulfilled all the criteria to be eligible for donation as described by the standard operating procedure of Uganda blood transfusion service. The participants' ages ranged between 15 and 51 years, and the median age was 18 years. The majority of the participants (>70%) were below 20 years of age because this regional blood bank repeatedly targets secondary school students to donate blood, most of whom happen to be below twenty (20) years of age. None of the blood donors were on immunosuppressive drugs or had a history of organ transplant. All blood samples from the blood donors enrolled in this study were tested for Hepatitis B surface antigen (HBsAg), anti-Hepatitis C, anti-Human Immunodeficiency Virus types 1 and 2 antibodies, and syphilis prior to HTLV-1/2 screening. The data extraction form was used to collect information regarding sociodemography, history of previous blood transfusion, blood donation, and level of education of the participant.Sociodemography information including sex, age in years, occupation, place of birth, and the participant's (blood donor) place of residence location were recorded. Study participants were identified by the study number and no names were included.n is minimum sample size required; Zα is standard normal deviate at 95% confidence interval corresponding to 1.96; P is assumed true population prevalence of HTLV-1/2 infections among blood donors at Mbarara Regional Blood Bank, South Western Uganda.The sample size was determined using the Kish  formula P is the probability of not having HTLV; δ is absolute error between the estimated and true population prevalence, Margin of Error (5%).Since true population prevalence of HTLV-1/2 infections among blood donors at Mbarara Regional Blood Bank was not known, a HTLV seroprevalence of 2.3% obtained from a similar study done among blood donors in Mozambique (the country nearest to Uganda where a similar study had been done) by Caterino-de-Araujo et al., 2010, was used as the estimated prevalence; 1 − One hasHowever, a total of 368 blood samples were screened. A decision to include 368 samples was made because up to this number (368 samples) could be analysed in a single run with the ELISA kit that was used.All donated blood was screened for Hepatitis B surface antigen, anti-Hepatitis C, anti-Human Immunodeficiency Virus types 1 and 2 antibodies, and syphilis at Mbarara Regional Blood Bank.All blood was submitted after the 368th sample.An enzyme-linked immunoassay was carried out for the determination of antibodies to HTLV-1 and HTLV-2 in serum and plasma using Murex HTLV-I + II DiaSorin, Dartford, UK (specificity of 99.5%), according to the manufacturer's recommendations. Screening of the blood samples for Hepatitis B surface antigen, anti-HCV antibodies, anti-HIV types 1 and 2 antibodies, and syphilis was done prior to anti-HTLV-1/2 testing. Initially reactive sera were confirmed by repeat testing and were considered positive only if they had double reactivity as the specificity of this test was high enough to confirm the infection.Data was double-entered in Microsoft Excel 2007 and was exported to Stata  for analysis.Univariate and bivariate analyses were done where appropriate. Descriptive statistical procedures were used to compute Chi-square for contingency tables.p value. The statistical significance was considered when the p value was below or equal to 0.05. Logistic regression was applied to estimate the magnitude of associations with risk factors for HTLV-1/2 seropositivity of the blood donors.The statistical inferences for the relationship between seropositivity and other factors were determined based on the Pearson Chi-square test of proportions' A total of three hundred and sixty-eight blood donors participated in this study: 229 (62.2%) were males and 139 (37.8%) were females . The donOf these 206 (55.98%) were first time donors, while 162 (44.02%) were repeat donors .The majority of the study participants were males who had attained a secondary level education.Of the 368 blood donors, two (2) were found to be seropositive for HTLV-1/2 antibodies giving a prevalence of 0.54% . None ofχ2  (4 df) = 0.0221, p = 1.0).The distribution of HTLV-1/2 seropositivity by community location or place of residence , education, and occupation showed that the two HTLV-1/2 seropositive blood donors were both from rural areas , had attχ2  (4 df) = 0.34, p = 0.99).All variables from the univariate analysis including age, sex, place of birth, community location of place of residence , occupation, and level of education were entered into multiple logistic regression models. In this analysis, no independent risk factors were found to be significantly associated with HTLV-1/2 seropositivity  is similar to that found among blood donors presenting to blood banks in Ghana  where blThe seroprevalence in this present study is comparatively low. Probably, this could be due to the fact that the majority of the participants in this study were secondary school students who are at a low risk for HTLV-1/2 infection while in the other studies , 22 therIn this study, all HTLV-1/2 seropositive donors were males. This is not in conformity with studies that have shown prevalence of HTLV-1 and HTLV-2 to be higher in females as opposed to males which is attributed to more efficient transmission from men to women during sexual intercourse .This is most likely due to the higher proportion of males compared with females in this study. This is a limitation of the study as gender of the participants was skewed in favour of the male sex.The two HTLV-1/2 seropositive blood donors were both from rural areas which were their places of origin as well. This finding is similar to that observed in studies done in Gabon and Mozambique by Le Hesran et al.  and Cate Sarcoptes scabiei var. Hominis as suggested by other studies has also been found to be associated with HTLV seropositivity among rural communities [Having a history of crusted scabies caused by massive infestation withmunities , 27.In this study it was not possible to further explore specific lifestyle factors associated with risk of acquiring HTLV infection in this group  due to the small proportion of seropositive donors from the rural area.p = 0.941), despite suggestions of such relations in previous studies [In this study, all HTLV-seropositive donors had attained education up to secondary level. Generally, the distribution of educational attainment was quite narrow and was almost identical between the seropositive and seronegative donors, perhaps reflecting a disadvantage of studying a more or less occupational based cohort. Thus, the study was unable to confirm any associations between HTLV-I/2 seropositivity and education, a marker for socioeconomic status ; therefore, the seroprevalence may have been underestimated and thus the result may not really be informative on an epidemiological and public heath level. Second, a higher proportion of males (62.2%) compared with females (37.8%) participated in this study. Since it is known that HTLV-1/-2 seroprevalence is higher in women than in men; the higher proportion of males in this study may also have led to an underestimation of the prevalence of HTLV-1/2 in females as opposed to males.Third, most samples were obtained from individuals less than 20 years old; this very likely led to an underestimation of the real HTLV-1/2 prevalence since it is known that HTLV-1/-2 seroprevalence is higher in older individuals than in younger ones. Finally, the reported HTLV-1/-2 prevalence was obtained using only a commercial EIA (specificity > 99.5%), and a confirmatory test (Western blot) or PCR was not done to confirm the results.Prevalence of HTLV 1/2 among blood donors at Mbarara Regional Blood Bank is too low to provide justification for screening of blood donations.


It has been reported that reactive oxygen species (ROS), such as hydrogen peroxide and superoxide, take part in osteoclast differentiation as intra-cellular signaling molecules. The current assumed signaling cascade from RANK to ROS production is RANK, TRAF6, Rac1, and then Nox. The target molecules of ROS in RANKL signaling remain unclear; however, several reports support the theory that NF-κB signaling could be the crucial downstream signaling molecule of RANKL-mediated ROS signaling. Furthermore, ROS exert cytotoxic effects such as peroxidation of lipids and phospholipids and oxidative damage to proteins and DNA. Therefore, cells have several protective mechanisms against oxidative stressors that mainly induce cytoprotective enzymes and ROS scavenging. Three well-known mechanisms regulate cytoprotective enzymes including Nrf2-, FOXO-, and sirtuin-dependent mechanisms. Several reports have indicated a crosslink between FOXO- and sirtuin-dependent regulatory mechanisms. The agonists against the regulatory mechanisms are reported to induce these cytoprotective enzymes successfully. Some of them inhibit osteoclast differentiation and bone destruction via attenuation of intracellular ROS signaling. In this review article, we discuss the above topics and summarize the current information available on the relationship between cytoprotective enzymes and osteoclastogenesis.  •Three essential regulatory mechanism for cytoprotective enzymes were reported.•They are Nrf2, SIRT, and FOXO.•Intracellular ROS is used as signaling molecule during osteoclastogenesis.•Cytoprotective enzymes attenuates osteoclastogenesis by ROS scavenging.•Nrf2, SIRT, and FOXO could be therapeutic target for bone destructive disease. Indeed, a well-known cytoprotective enzyme, HO-1, is a negative regulator of osteoclastogenesis Another mechanism regulating cytoprotective enzyme FOXO contributes to the control of osteoclastogenesis. Bartell et al. reported that FOXO protein attenuates osteoclastogenesis via augmentation of cytoprotective enzymes 5As discussed above, osteoclasts also possess mechanisms that regulate cytoprotective enzymes, which manage the intracellular ROS levels. Since intracellular ROS play a role in RANKL-mediated osteoclastogenesis, the mechanisms that regulate cytoprotective enzymes negatively control osteoclastogenesis via ROS scavenging mediated by cytoprotective enzymes. In other words, osteoclastogenesis is controlled via interference with the mechanisms regulating cytoprotective enzymes.Indeed, some papers report that the activation of Nrf2 inhibits osteoclastogenesis 6In this review manuscript, we have summarized recent information about the relationship between osteoclastogenesis and the mechanisms regulating cytoprotective enzymes. Although some parts have been extensively explored, further investigations are necessary to gain a greater understanding. In particular, crosstalk among the mechanisms regulating cytoprotective enzymes and other signaling molecules should be elucidated.Since some of the agonists that affect the mechanisms regulating cytoprotective enzymes have been reported as inhibitors of bone destruction, these chemicals could be potential drugs for the treatment for bone destructive diseases in the near future.


Analysing total KIR expression showed the presence of KIR+CD8+ T cells, NK cells and CD4+ T cells in PBMCs from uninfected and HTLV-1+ donors. A subset of HTLV-1+ donors had high frequencies of total KIR+CD8+ T cells. Analysing individual KIRs revealed the presence of KIR2DL2+CD8+ T cells in HTLV-1+ donors. Preliminary data from PBMCs stimulated with Tax peptides indicates that KIR2DL2+CD8+ T cells constitute a very small proportion of the IFNγ-producing Tax-specific CD8+ T cell population. HTLV-1+ asymptomatic carriers had higher frequencies of IFNγ-producing Tax-specific KIR2DL2+CD8+ T cells than donors with HAM/TSP did. Further work is underway to characterise the function of Tax-specific CD8+ T cells by staining with anti-CD107a, anti-IFNγ and the HLA-A2/Tax11-19 pentamer, and to compare the frequency and function of KIR2DL2+CD8+ T cells with those expressing other 2-domain KIRs.An efficient antiviral cytotoxic T lymphocyte (CTL) response to HTLV-1 infection maintains a low proviral load (PVL), reducing the risk of HAM/TSP. Host genotype, particularly of HLA class I, is a major determinant of CTL efficiency, and the influence of specific HLA class I alleles on HTLV-1 immunity is well documented. We recently showed that killer immunoglobulin-like receptor (KIR) genotype also influences CTL efficiency, by affecting HLA class I-mediated HTLV-1 immunity. Possession of the KIR2DL2 gene enhanced the effect of known protective or detrimental HLA class I alleles on PVL and HAM/TSP risk. This study aims to profile the frequency and function of CD8


Malignant pleural mesothelioma (MPM) is a cancer associated with exposure to asbestos fibers, which accumulate in the pleural space, damage tissue and stimulate regeneration. Hedgehog signaling is a pathway important during embryonic mesothelium development and is inactivated in adult mesothelium. The pathway is reactivated in some MPM patients with poor clinical outcome, mainly mediated by the expression of the ligands. Nevertheless, mutations in components of the pathway have been observed in a few cases. Data from different MPM animal models and primary culture suggest that both autocrine and paracrine Hedgehog signaling are important to maintain tumor growth. Drugs inhibiting the pathway at the level of the smoothened receptor (Smo) or glioma-associated protein transcription factors (Gli) have been used mostly in experimental models. For clinical development, biomarkers are necessary for the selection of patients who can benefit from Hedgehog signaling inhibition. BAP1) predispose individuals to MPM [Malignant pleural mesothelioma (MPM) is an aggressive tumor arising from the mesothelial lining cells of the pleura . MPM is s to MPM .MPM is a very heterogeneous and highly chemoresistant tumor; therefore, management remains a clinical challenge. The best response rates are being reported after chemotherapy with cisplatin and pemetrexed . SeveralBest survival data are reported from groups combining macroscopic complete resection achieved by extrapleural pneumonectomy or pleurectomy/decortication, chemo- and radio-therapy in a multimodal treatment ,11,12,13For patients who cannot benefit from multimodality treatment, other treatment options, including immunotherapy, as well as targeting oncogenic pathways activated in MPM are under evaluation in the clinic (reviewed in ). One paGli1 and Hedgehog interacting protein (Hhip). The latter competes with Ptch by binding to Hh ligands [Classical Hh core signaling components (reviewed in ) are the ligands . Gli pro ligands ,22. In t ligands .SHH gene expression in human MPM tumor tissues along with increased expression levels of HHIP and GLI1 [GLI1 gene expression was associated with shorter overall survival in MPM patients [The Hh signaling pathway is essential for embryonic mesothelial development . In the and GLI1 . High GLpatients . Anotherpatients .http://gdac.broadinstitute.org/). The pipelines run in a computational framework called Firehose, which also generates analysis reports [GLI1, HHIP and PTCH2. For 39 of these patients, clinical outcome is also available, and the Hh target gene-expressing group is one of the two groups with the worst overall survival . Since 2006, TCGA has explored these differences in many cancer types using a variety of platforms using single-nucleotide polymorphism, small RNA transcriptome, exome and methylation data from sequencing and microarrays. Systematic analysis of TCGA datasets was generated through the Broad’s Genome Data Analysis Center .Altogether, published work and data mining indicate that deregulated Hh signaling defines a subgroup of patients with bad clinical outcome.SUFU gene, leading to T411M amino acid change, coupled with three base pair (CTG) insertions in the SMO gene, resulting in an additional amino acid 23L_24GinsL in the signal peptide region in one cell line [SMO was also detected in one MPM patient out of 14 patients analyzed. Transfection of a SUFU cDNA harboring the T411M missense mutation suppressed Gli-reporter gene downregulation as observed with wild-type SUFU [PTCH1 exons 8–23 was observed in another cell line [PTCH1 gene, is observed in medulloblastoma and basal cell carcinoma  and variewed in ,33,34, cPTCH1 expression compared to the non-side population [As mentioned above, the Hh pathway is important for development and tissue repair, and it is also active in cancer cells with the stem cell phenotype . We pulation .in vitro [Stem cell signaling is, as also observed by others , not maiin vitro  and reduin vitro . Thus, tIn addition, we recently detected paracrine activation of Hh signaling in MPM. Analysis of GLI1 immunoreactivity in a small cohort of MPM patients revealed heterogeneous expression in both tumor and stroma fractions . In ordeA number of clinical trials for Hh inhibitors, mainly Smo antagonists, have been conducted to date . Thrin vitro [Several lines of evidence suggest existing crosstalk between Gli1 and other signal transduction pathways (reviewed in ) in an Hin vitro ,63,64.In summary, several lines of evidence indicate that Hh signaling is activated in a subgroup of MPM patients and is associated with the worst clinical outcome. In a limited number of them, activation may result from either mutation in components of the pathway, while for the others, it seems to be resulting from ligand-driven activation. The identification of biomarkers and parallel or compensatory non-canonical signaling may be useful for exploring mechanism-based therapeutic combinations.


The development of stable tissue-engineered autologous bone grafts in the field of regenerative medicine is still a challenge. Perfusion bioreactors not only provide continuous nutrition supply and waste removal, but are also suitable for the controlled application of mechanical forces like fluid shear stress. Mechanical loading is known to cause mechanotransductive effects like the induction of differentiation, resulting in enhanced deposition of extracellular matrix .In our study, we determined the optimal flow rate for the osteogenic differentiation of human adipose-derived mesenchymal stem cells (MSC) by applying fluid shear stress that mimics the physiological environment normally experienced by bone progenitor cells in vivo. For this, we first analyzed the porosity of cell substrates with nanofocus-computed tomography as well as their specific permeability at different flow rates. To investigate the effect of controlled application of physiologic fluid shear stress a flow rate of 0.3 ml/min was used to cultivate MSC in a self-developed perfusion bioreactor. Cells were seeded on a three-dimensional macro-porous zirconium dioxide ceramic scaffold (0.3·106 cells/scaffold) and cultivated in standard growth medium (GM)or osteogenic differentiation medium (ODM) under normoxic (21% O2) or hypoxic (5% O2) conditions for a period of 21 days. After cultivation cell viability was examined using MTT assay. Furthermore DAPI staining was used to evaluate cell distribution. Glucose consumption and lactate production were monitored and histological stainings  were used to evaluate osteogenic differentiation.Flow rates between 0.3 - 5 ml/min result in fluid shear stress between 0.01 - 2.5 Pa which is in the range of physiologic shear stress bone progenitor cells are subjected to in vivo (0.3 - 3 Pa)[Physiologic fluid shear stress together with physiologic oxygen conditions (5% O2) lead to higher cell viability. Furthermore the glucose metabolism is elevated under perfusion due to enhanced mass transfer. The application of fluid shear stress results in a stronger differentiation regarding matrix deposition even in standard growth medium without any osteoinductive supplements. These results underline the positive effects of dynamic cultivation and physiologic oxygen concentration which together mimic in vivo conditions. Consequently other factors like medium composition should be adjusted to be more physiologic and taken into consideration during a tissue engineering process to ensure a physiologic tissue maturation.Part of this work was performed with financial support of the FFG BRIDGE project "3D Tissue".


SETBP1 mutations, which arepresent in 24.3% of aCML patients. Moreover, both shared the variant encodingp.Pro737His, but the aCML severity was greater in the son because of the presence oftwo other missense mutations causing p.Asp868Asn and p.Ser885Arg alterations.SETBP1 mutations may be associated with an adverse prognosis, sotheir detection would help in the diagnosis of aCML and the determination of apatient's prognosis.We report the case of a father and son diagnosed with atypical chronic myeloidleukemia (aCML). Both patients harbored  BCR-ABL fusion gene.Atypical chronic myeloid leukemia (aCML), a rare disorder of hematopoietic stem cells,has both myelodysplastic and myeloproliferative characteristics and is classified as anexample of myelodysplastic/myeloproliferative neoplasms (MDS/MPN) according to the WorldHealth Organization (WHO) classification system . aCML shSETBP1 encodes SET-binding protein 1, a binding partner for themulti-functional SET protein, which is involved in apoptosis, transcription, andnucleosome assembly. A 2013 analysis of exome sequences from 8 aCML cases led to theidentification of recurrent SETBP1 somatic mutations; then targetedresequencing demonstrated the SETBP1 mutations were present in 24.3% of70 patients with aCML, as well as in the closely related disorders unclassified MDS/MPN and chronic myelomonocytic leukemia , and in one of fourcases of chronic neutrophilic leukemia (CNL) (CSF3R), most commonlyCSF3R T618I  in the bonemarrow sample.The 30-year-old son was admitted to our hospital on July 2, 2013, because of an enlargedspleen with a maximum thickness of 9.6 cm. His peripheral blood count status was: whiteblood cells (WBCs), 16.75×10feration , and conSETBP1 mutations  identified three missense mutationscausing p.Pro737His, p.Asp868Asn, and p.Ser885Arg alterations  was administered for 14 days each month beginningJuly 11, 2013, and interferon α-2a (300 MU) was administered three times weekly bysubcutaneous injection but was discontinued because of intolerance after severaladministrations. However, no significant improvement in fatigue, spleen size, or bonemarrow blasts occurred, and the patient suffered aggravated anemia and persistent feverwhile being treated with antibiotics. On September 13, 2013, the patient began treatmentwith imatinib mesylate (400 mg/day) , whichw9/L; hemoglobin, 115 g/dL; and platelets, 1190×109/L.Ultrasonography and computerized tomography demonstrated a normal liver but an enlargedspleen with a maximum thickness of 8.3 cm, and 7.4 cm under the left rib cage. Tests forBCR/ABL JAK2 617F and CSF3R mutations were negative, although the bone marrow smear andbiopsy were indicative of aCML. He was treated with interferon α-2a (3.0 MU/day) pluslow-dose cytarabine (20 mg/m2 during days 1-10) for approximately 4 months.The regimen was voluntarily discontinued after the symptoms and peripheral blood countstatus improved. In August 2013, the father was alive and stable, with no indication ofdisease progression such as fever, anemia, increased blasts, or an enlarged spleen.Analysis of SETBP1 mutations in DNA from peripheral blood revealed thepresence of the variant encoding p.Pro737His, which was shared with his son  and with the HelsinkiDeclaration of 1975, as revised in 2000   and presented with higher WBC counts at the time of diagnosis  compared with aCML cases with wild-typeSETBP1. Another team showed that SETBP1mutationsoften co-occurred with cytogenetic markers such as -7/del(7q) and i(17)(q10), which wereassociated with a reduced overall survival time and an increased risk of leukemicevolution from MDS  -13, assfrom MDS .SETBP1 mutations was ineffective atdifferentiating aCML from CNL. CSF3R encodes the cell surfacetransmembrane receptor for granulocyte colony stimulating factor, which induces theproliferation, differentiation, and survival of myeloid progenitors. In 2013, Maxson etal.  and aCML . Two types ofCSF3R mutation were found: T618I (n=12) and T615A (n=2), which areboth located in the membrane proximal region that mediates proliferative and survivalsignals. Pardanani et al.  or aCML (n=19). A central pathology review identified 12 WHO-defined CNL and 9WHO-defined aCML patients. CSF3R T618I mutations were observed in 10WHO-defined CNL cases, at a mutational frequency of 83% (10/12), and were not seen inWHO-defined aCML, PMF (n=76), or CMML (n=94) cases. A further 3 non-CNL cases alsoharbored CSF3R mutations.However, the presence of ML 8/18, 4.4%. TwoCSF3R mutation positivity (i) a peripheral blood leukocyte level ≥13×109/L,ii) a peripheral blood neutrophil/band percentage distribution>80%, and iii) the presence of a CSF3R T618Imutation or other membrane proximal CSF3R mutation. Therefore, althoughmost CNL patients carry SETBP1 and ASXL1 mutations,which can also be detected in aCML, CSF3R mutations, particularlyCSF3R T618I, can be used to differentiate them.Together, these findings ensure that the diagnosis of CNL is no longer only one ofexclusion, and revision of the current WHO diagnostic criteria is expected to includethe molecular criterion of sitivity . In 2014sitivity ,6 proposSETBP1 mutations canco-occur in a father and son with aCML, but it remains to be determined whether thesewere inherited or acquired. The detection of SETBP1 andCSF3R mutations will play an important role in diagnosing aCML, CNL,and CMML patients, and determining their prognoses. These molecular markers are likelyto be incorporated into new diagnostic criteria soon.In the present cases, we demonstrated that


Human T-cell leukemia virus type 1 (HTLV-1) is a causative virus of adult T-cell leukemia (ATL), and inflammatory diseases. HTLV-1 bZIP factor (HBZ) is encoded on the minus strand of HTLV-1, and expressed in all ATL cases. We performed RNA-seq and ChIP-seq using HBZ transduced T cells, and found that expressions of several Treg-related genes, including Foxp3, CD25, CCR4, CCR5, PD-1, NRP1, IKZF family genes, and T-cell immunoglobulin and ITIM domain receptor (Tigit), were upregulated by HBZ. We focused on TIGIT in this study. TIGIT expression was also upregulated in HTLV-1 infected cells in HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) patients and CD4+ T cells of ATL patients. HBZ induced the histone modification of the promoter of TIGIT and enhanced its expression. HBZ also enhanced acetylation of H3K18, which is the specific target of p3. HBZ augments transcription of Tigit gene in the presence of PMA/Ionomycin stimulation. Although the detailed mechanism of HBZ mediated activation of Tigit transcription is still unclear, p3 and activated pathway by PMA/Ionomycin is likely involved in its regulation. Furthermore, we analyzed the function of Tigit in HBZ-expressing T cells and CD4+ T cells from HBZ-transgenic mice (HBZ-Tg), and found that stimulation of Tigit with its ligand, PVR, enhanced expression of inhibitory cytokines, such as IL-10, in HBZ-Tg. These results suggest that the enhanced Tigit expression by HBZ may play a key role to modulate the microenvironment where anti-tumor immune response is attenuated by increased expression of IL-10. This environment seems to be associated with the evasion of HTLV-1 infected cells from anti-tumor host immune response and the pathogenesis of HTLV-1 associated inflammatory diseases and ATL.


Toll-like receptors (TLRs) play crucial roles in the innate immune system by recognizing pathogen-associated molecular patterns derived from various microbes. TLRs signal through the recruitment of specific adaptor molecules, leading to activation of the transcription factors NF-κB and IRFs, which dictate the outcome of innate immune responses. During the past decade, the precise mechanisms underlying TLR signaling have been clarified by various approaches involving genetic, biochemical, structural, cell biological, and bioinformatics studies. TLR signaling appears to be divergent and to play important roles in many aspects of the innate immune responses to given pathogens. In this review, we describe recent progress in our understanding of TLR signaling regulation and its contributions to host defense. The innate immune system employs germline-encoded pattern-recognition receptors (PRRs) for the initial detection of microbes. PRRs recognize microbe-specific molecular signatures known as pathogen-associated molecular patterns (PAMPs) and self-derived molecules derived from damaged cells, referred as damage-associated molecules patterns (DAMPs). PRRs activate downstream signaling pathways that lead to the induction of innate immune responses by producing inflammatory cytokines, type I interferon (IFN), and other mediators. These processes not only trigger immediate host defensive responses such as inflammation, but also prime and orchestrate antigen-specific adaptive immune responses . These rMammals have several distinct classes of PRRs including Toll-like receptors (TLRs), RIG-I-like receptors (RLRs), Nod-like receptors (NLRs), AIM2-like receptors (ALRs), C-type lectin receptors (CLRs), and intracellular DNA sensors such as cGAS . Among tTLRs are expressed in innate immune cells such as dendritic cells (DCs) and macrophages as well as non-immune cells such as fibroblast cells and epithelial cells. TLRs are largely classified into two subfamilies based on their localization, cell surface TLRs and intracellular TLRs. Cell surface TLRs include TLR1, TLR2, TLR4, TLR5, TLR6, and TLR10, whereas intracellular TLRs are localized in the endosome and include TLR3, TLR7, TLR8, TLR9, TLR11, TLR12, and TLR13 . TLR2 along with TLR1 or TLR6 recognizes a wide variety of PAMPs including lipoproteins, peptidoglycans, lipotechoic acids, zymosan, mannan, and tGPI-mucin . TLR5 rePlasmodium falciparum during the process of detoxification after host hemoglobin is digested (Escherichia coli (UPEC) as well as a profilin-like molecule derived from Toxoplasma gondii P on endosomes, which mediates the formation of functional TLR4 and TLR9 signaling complexes at their respective sites. Thus, TIRAP associates with both cell surface and endosomal TLRs by binding to different lipids . Du. Du42). TAK1 deficiency in mouse embryonic fibroblast cells (MEFs) reduces phosphorylation of IKKs, p38, and JNK after LPS stimulation. However, TLR4-mediated IKK, p38, and JNK activation and cytokine induction are increased in neutrophils derived from TAK1-deficient mice, suggesting a cell type-specific role for TAK1 in TLR signaling . FurtherTLR2 and TLR4 ligations in macrophages increase the production of mitochondrial ROS for bactericidal action and recruit mitochondria to phagosomes . TRAF6 iTRIF interacts with TRAF6 and TRAF3. TRAF6 recruits the kinase RIP-1, which in turn interacts with and activates the TAK1 complex, leading to activation of NF-κB and MAPKs and induction of inflammatory cytokines Figure , 5.The Pellino family E3 ubiquitin ligases are implicated in TLR signaling . PellinoRecently, IRF3 activation was demonstrated to be regulated by an inositol lipid, PtdIns5P. PtdIns5P binds to both IRF3 and TBK1, and thus facilitates complex formation between TBK1 and IRF3. The accessibility of TBK1 to IRF3 mediated by PtdIns5P likely causes IRF3 phosphorylation in a closely proximal manner. Furthermore, PIKfyve was identified as a kinase responsible for production of PtdIns5P during virus infection .TLR4 activates both the MyD88-dependent and TRIF-dependent pathways. Activation of these pathways is controlled by several molecules to induce appropriate responses. Balanced production of inflammatory cytokines and type I IFN may be important for controlling tumor cell growth and autoimmune diseases.TRAF3 was shown to be incorporated into the MyD88 complex as well as the TRIF complex in TLR4 signaling. TRAF3 within the MyD88 complex is then degraded, which causes TAK1 activation. Thus, in addition its role in promoting TRIF-dependent pathway activation, TRAF3 has a role in inhibiting the MyD88-dependent pathway. NRDP-1, an E3 ubiquitin ligase, binds and ubiquitinates MyD88 and TBK1, inducing the degradation of MyD88 and augmenting the activation of TBK1, which attenuates inflammatory cytokine production and induces preferential type I IFN production, respectively .MHC class II molecules that are localized in endosomes in antigen-presenting cells interact with the tyrosine kinase Btk via the costimulatory molecule CD40 and maintain Btk activation. Activated Btk interacts with MyD88 and TRIF to promote the activation of the MyD88-dependent and TRIF-dependent pathways and thus to enhance production of inflammatory cytokines and type I IFNs, respectively .Plasmacytoid DCs are a subset of DCs with the capacity to secrete vast amounts of type I IFN in response to viral infection Figure  2, 5). , 5. 2, 5In addition to IRF3 and IRF7, several other IRFs participate in TLR signaling. IRF1 interacts with MyD88 and contributes to TLR9-mediated cytokine production in the presence of IFNγ , while IRecent studies have identified several transmembrane molecules that modulate TLR signaling pathways. CD14, a glycophosphatidylinositol-anchored protein, is a co-receptor with TLR4 and MD-2 for LPS recognition. It induces ITAM-mediated Syk- and PLCγ2-dependent endocytosis to promote TLR4 internalization into endosomes for activation of TRIF-dependent signaling . CD14 isCD36, a protein in the class B scavenger receptor family, acts as a co-receptor for oxidized low-density lipoprotein (LDL) and amyloid-β peptide. Ligand recognition induces the assembly of TLR4/TLR6 heterodimers through Src kinases and consequent sterile inflammation, by inducing inflammatory cytokines and ROS and priming NLRP3 inflammsome activation , 68.TLR signaling is negatively regulated by a number of molecules through various mechanisms to prevent or terminate the excessive immune responses that lead to detrimental consequences associated with autoimmunity and inflammatory diseases. Negative regulators target each of the key molecules in TLR signaling Figure . ActivatDuring the past decade, tremendous progress has been made in our understanding of TLR signaling pathways. After genetic studies revealed the contribution of TIR domain-containing adaptor usage, cell biological and biochemical approaches have highlighted the importance of cellular localization of these adaptors in the regulation of downstream signaling. Moreover, numerous reports have demonstrated that TLR trafficking, TLR cleavage, and protein modification of signaling molecules such as ubiquitination and phosphorylation play important roles in the activation of TLR signaling. On the other hand, negative regulators of TLR signaling have been discovered, and their importance in preventing autoimmune and inflammatory diseases is recognized. More recently, much effort has been focused on identifying molecules that are involved in innate immunity through an integrated approach. Indeed, by combining transcriptomics, genetic/chemical perturbations and phosphoproteomics, Polo-like kinases (Plks) 2 and 4 have been found to regulate antiviral responses downstream of TRIF and MyD88 signaling . mRNA stAlthough PAMP recognition by TLRs is crucial for host defense responses to pathogen infection, aberrant activation of TLR signaling by PAMPs, mutations of TLR signaling molecules, and DAMPs-mediated TLRs signaling activation are responsible for the development of several diseases such as autoimmune, chronic inflammatory, and allergic diseases. Moreover, a link between cancer and TLRs has been proposed. The innate immune activation that caused after anti-cancer drug treatment is reportedly critical for cancer elimination through TLR-mediated recognition of endogenous molecules released from dying cancer cells . On the 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.


Vibrio coralliilyticus is a pathogen of corals and larval shellfish. Publications on strain RE98 list it as a Vibrio tubiashii; however, whole genome sequencing confirms RE98 as V. coralliilyticus containing a total of 6,037,824 bp consisting of two chromosomes  and two megaplasmids . Vibrio coralliilyticus is a well-known coral pathogen responsible for coral bleaching and the associated losses to coral reefs worldwide (Crassostrea gigas) larvae (C. virginica) larvae is Vibrio tubiashii, which causes high mortalities in oyster and clam hatcheries and potentially in the wild  larvae , 4. Anotthe wild –8. Some s tested . The misV. coralliilyticus RE98 was sequenced using a PacBio RS II system  on single-molecule real-time (SMRT) cells using PacBio P5-C3 chemistry. Subread filtering was performed with the SMRT Analysis Software suite  and revealed 5,718 genes, 5,467 coding sequences, 98 pseudogenes, 34 rRNAs , 116 tRNAs, 3 noncoding RNA, and 11 frameshift genes. A Blast search of chromosomes 1 and 2 using the NCBI whole-genome shotgun (WGS) contig database, limited by organism (Vibrionaceae), showed >97% sequence similarity to other V. coralliilyticus.Genome annotation for V. coralliilyticus RE98 (chromosomes 1 and 2 and its two megaplasmids) has been deposited in GenBank under accession no. CP009617, CP009618, CP009619, and CP009620.The complete genomic sequence of


HTLV-1 is responsible for two main diseases, Adult T-cell Leukemia/Lymphoma and HTLV-1 Associated Myelopathy/Tropical Spastic Paraparesis, for which there is currently no satisfactory treatment. Among the proteins encoded by HTLV-1, Tax appears to play an important role in the mechanisms leading to pathogenicity. We are interested in the mechanisms of cell transformation by Tax and more particularly in the interplay between the viral Tax oncoprotein and the DNA damage response (DDR). We demonstrated that transient expression of Tax results in DNA damage, cell cycle arrest and activation of the DDR. In fibroblasts, cell cycle arrest occurs at the G1 and G2 phases depending on the p53 background. Although Tax induces apoptosis and senescence in fibroblasts, HTLV-1 infected lymphocytes proliferate continuously and appear to be adapted to the checkpoint control. This mechanism allows infected lymphocytes to proliferate despite the presence of genomic lesions. With these observations, we propose a novel therapeutic approach based on the principle of synthetic lethality.


HTLV-1 infection may is present among HCV or HIV-infected subjects and is associated with higher risk for HAM/TSP and other inflammatory diseases. Lymphocytes from about half of HTLV-1-infected subjects spontaneously proliferate in vitro, and how this phenomenon relates to symptomatic disease outcome and viral burden is poorly understood. Objective: Evaluate T-cell proliferation in vitro and HTLV-1 proviral load (PVL) among co-infected subjects.From 522 HTLV-1-infected individuals, who presented co-infection with HCV or HIV were selected during their outpatient visits to the HTLV clinic at the IIER in Sao Paulo. All the volunteers were invited to participate after reading and signing a consent form. Clinical data were obtained from medical records and interviews. PBMC from patients and controls, 2 x106 cells/ml in RPMI with 10% fetal calf serum were incubated at 37 C and 5% CO2 for 3 days in triplicate. Cells were pulsed with tritiated thymidine 18 h before harvest in a semi-automatic cell harvester and counted in a beta-counter. To determine the proviral load, the HTLV-1 DNA copy number was done by RT-PCR to the amount of the cellular albumin of the clinical sample, which was quantified in parallel. Results were expressed as HTLV-1 DNA copies/104 PBMCs. Statistical analysis was conducted using Kruskal-Wallis test.From a total infected with HTLV-1, 123 (24%) subjects presented co-infection, where 81 had HIV-1 (15%), 40 had HCV (8%) and 37 had HIV-1 and HCV (7%). The basal T-cell proliferation show an increase among HCV/HTLV-1 subjects compared to patients with HTLV-1/HIV-1 co-infection and HTLV-1/ HIV-1/HCV (p=0.1). The HTLV-1 PVL was also highest in the HTLV-1/HCV subjects (p=0.6).We observed a significant increase of basal T-cell proliferation among HTLV-1/HCV co-infected. However, the presence of co-infection with HIV-1 may induce a down regulation at T-cell proliferation capacity. Support: FAPESP 2012/23397-0.


The canonical ERK-MAPK signaling cascade  has been implicated in controlling diverse cellular behaviors, including proliferation and differentiation. While the molecular interactions involved in signal transduction through this cascade have been well characterized in cell culture experiments, our understanding of how this sequence of events unfolds to determine cell fate within a homeostatic tissue environment has not been fully characterized.The skin is largely comprised of keratinocytes within the interfollicular epidermis. Over approximately two weeks these cells differentiate and traverse the thickness of the skin. The stage of differentiation is therefore reflected in the positions of cells within the tissue, providing a convenient axis along which to study the signaling events that occur We measured the abundance of total and phosphorylated ERK-MAPK signaling proteins within interfollicular keratinocytes in transverse cross-sections of human epidermis using immunofluorescence microscopy. To investigate these data we developed a mathematical model of the signaling cascade using a normalized-Hill differential equation formalism.These data show coordinated variation in the abundance of phosphorylated ERK-MAPK components across the epidermis. Statistical analysis of these data shows that associations between phosphorylated ERK-MAPK components which correspond to canonical molecular interactions are dependent upon spatial position within the epidermis. The model demonstrates that the spatial profile of activation for ERK-MAPK signaling components across the epidermis may be maintained in a cell-autonomous fashion by an underlying spatial gradient in calcium signaling.in situ, and statistical associations in these data indicate canonical ERK-MAPK interactions underlie this spatial profile of ERK-MAPK activation. Using mathematical modelling we have demonstrated that spatially varying calcium signaling components across the epidermis may be sufficient to maintain the spatial profile of ERK-MAPK signaling cascade components in a cell-autonomous manner. These findings may have significant implications for the wide range of cancer drugs which therapeutically target ERK-MAPK signaling components.Our data demonstrate an extended phospho-protein profile of ERK-MAPK signaling cascade components across the epidermis The online version of this article (doi:10.1186/s12918-015-0187-6) contains supplementary material, which is available to authorized users. Sigin vitro , integriin vitro , 22 and in vitro , 23. Thein vitro , 24. Thein vitro , 26.in vitro models [Drosophila and other malleable systems [in situ. Furthermore, cells grown in vitro are exposed to the different environmental cues than cells in their in vivo tissue, and this can have significant effects on phenotypic behavior and intracellular signaling. For example, epidermal keratinocytes grown in vitro undergo abnormal differentiation expressing proteins more commonly associated with wound-healing and basal cell carcinoma [The ERK-MAPK cascade has been extensively characterized using o models –30 and co models , 31–33, o models  or some o models . A numbe systems , 35. Wit systems –38, raisarcinoma –41.in situ, by measuring abundance of signaling pathway components and mathematically modeling the spatiotemporal dynamics of the signaling cascade. We used immunofluorescence to measure the abundance of phospho-Raf-1, phospho-MEK-1/2, phospho-ERK-1/2 and calmodulin (CaM) within healthy human skin samples. A keratinocyte’s position within the epidermis, along the deep-to-superficial axis of the tissue, directly reflects its relative stage of differentiation. We used this to develop a computational framework for transforming the image data into a quantitative format, which allowed us to combine observations across multiple experiments. Our results show gradual, coordinated increases in the abundance of ERK-MAPK phospho-proteins over the depth of the skin, over much longer timescales than those observed for in vitro studies. Statistical analyses highlighted the importance of considering spatial position within the tissue when examining relationships between the measured variables and suggested canonical interactions are active during keratinocyte differentiation. Thus, we constructed a mathematical model of ERK-MAPK signaling interactions derived from the literature, using a normalized-Hill differential equation approach [2+ signaling components which vary over the depth of the epidermis were sufficient to drive signaling through canonical ERK-MAPK interactions and reach different steady-state activation levels, in a manner which was consistent with the extended spatiotemporal phosphorylation profiles observed within our data. Our results suggest that canonical ERK-MAPK interactions are operating, as overall pathway activity is modulated through the depth of the epidermis.In this study we therefore determined to characterize ERK-MAPK signaling in human epidermis approach , 43, witz-positions and tracked input co-ordinates through ‘mouse clicks’. This provided a relatively quick approach to sample the image data across multiple patients and targets, while distinguishing distinct subcellular domains of the cytoplasm, nucleus  and superficial (d2) boundaries of each tissue layer , in the absence of any spatial information.Our image processing framework transforms fluorescence intensity data onto the layer-normalized distance ordinate Fig. , producip-value ≤0.01). Many are well-studied, canonical signaling interactions, suggesting that these relationships may be active over the gradient of keratinocyte terminal differentiation .The signaling network  through the depth of the epidermis . In particular, the immunofluorescence data showed large decreases in the abundance of cytoplasmic phospho-Raf-1 and both cytoplasmic and nuclear phospho-MEK-1/2, but the model simulated little or no decrease in the phosphorylated abundance of these species. This can probably be attributed to the degradation processes activated during the final stages of keratinocyte differentiation [2+ and plasma-membrane CaM, and at each position in the epidermis we solve our simple model of the ERK-MAPK cascade within keratinocytes to steady-state. Modulation of ERK-MAPK phosphorylation over these extended timescales may play a central role in the pattern formation processes which establish and maintain epidermal homeostasis.Agreement between our immunofluorescence data and model predictions over the spinous and granular layers suggests that the network structure of the canonical signaling cascade with the specified feedback loops is sufficient to reproduce key features of the extended spatial phosphorylation gradient of ERK-MAPK components observed in human skin. As differentiating keratinocytes progress through the depth of the epidermis and change their expression of signaling receptors and adhesion molecules, the activity of signaling pathways within the keratinocytes shifts, and settles to a new, local steady-state of activation. In our model, we represent tissue microenvironment changes through the relative abundances of tissue Cain vitro cell signaling where there is only random heterogeneity across the spatial domain.In this study, we developed a novel computational approach to quantify ERK-MAPK activation within human epidermis. Our layer-normalized ordinate incorporates histological landmarks which separate distinct epidermal tissue layers. This allowed us to combine protein abundances measured using single-target labeling into an aggregate data set for mathematical modeling. Previous studies have used the relationship between a keratinocyte’s position within the epidermis and its stage of differentiation, examining protein abundance data  with respect to the position within the epidermis, and constructing ‘quantitative spatial profiles’ , 48. Immi.e. there is a relatively high mutual information between the abundance of successive phosphorylated pathway components). The regulation of signaling component compartmentalization is also thought to be critical for specificity of intracellular signaling pathway responses to different stimuli [For cell culture experiments a high degree of intercellular heterogeneity has been noted for ERK signaling, such that the population mean can provide a poor representation of the signal within individual cells . Our sta stimuli , 51, thue.g. phosphorylation) used default parameters, and only four parameters were optimized: the baseline levels and amplitudes of fractional activation for the input Ca2+ and CaM  can reproduce general features of the immunofluorescence data measured in situ, when driven by underlying changes in the surrounding tissue microenvironment. In particular, the relative abundances of tissue Ca2+ and plasma-membrane CaM were implemented as drivers of the model with spatial gradients, suggesting these molecules may be sufficient to establish the extended ERK-MAPK phosphorylation gradient we observed within homeostatic human epidermis. This is largely consistent with observations that increasing extracellular Ca2+ ion concentration promotes keratinocyte terminal differentiation in vitro [2+ gradient  and Counties-Manukau District Health Board (project number 681).Fresh skin samples obtained from healthy patients undergoing plastic or reconstructive surgery were snap frozen in liquid nitrogen and stored at −80 °C.A detailed description of the immunofluorescence labeling and confocal microscopy protocols, including antibody catalogue numbers, are given elsewhere . The antHuman epidermis was sectioned within a cryostat at −20 °C and immediately fixed with paraformaldehyde at room temperature, then labeled by indirect immunofluorescence and imaged using a Leica TCS SP2 confocal microscope. Secondary antibody control slides were imaged at the same gain and offset settings to ensure that the effects of non-specific labeling or auto-fluorescence were minimal. An imaging resolution at or exceeding the Nyquist sampling criteria was used, allowing the plasma-membrane, nucleus and cytoplasm of individual cells to be identified. Gain and offset values were adjusted to minimize underflow and overflow, and under these imaging conditions, fluorescence signal intensity was interpreted as an approximately linear measure of protein abundance within each voxel . Image dz-score normalized for each patient and then combined.A collection of scripts for quantifying the image data was written using MATLAB . A graphical user interface was developed using the MATLAB image processing toolbox and the ginput2 function  to facilitate selection of regions of interest (ROIs). Tissue layer boundaries were specified using the path tool within GIMP (v. 2.4.7) and exported as a binarized .TIFF format. The minimum distance between ROIs and surrounding tissue layer boundaries was determined using the intrinsic MATLAB function dist. For analyses where data were combined across patients Fig. , the sigLoess smoothing was chosen due to its ability to cope with edge effects  . Loess s“the strength of statistical association between measured fluorescence signals  is independent of spatial conditioning”Image data  100,000 times. From these scrambled data, the Pearson’s correlation and mutual information were calculated between targets and the probability density functions were taken as null distributions. The two-sided (Pearson’s correlation) and one-sided  99 % confidence intervals were identified as thresholds .Signal intensity data were extracted over 28 spatial partitions Table . Data wez-positions through the image data. A series of image processing functions were applied to: connect these data points and fill the selected area, smooth edges and subtract the nucleus from the cytoplasm, and interpolate the segmented areas between z-positions to produce a volumetric segmentation. From these data, the volume ratio of cytoplasm-to-nucleus was calculated for a number of cells across the depth of the epidermis, as shown in Additional file Scripts were written using MATLAB to manually input sample points that demarcate the edge of cellular domains at various http://sourceforge.net/projects/EpidermalERKMAPK/.An SBML  implemen2+ is a direct input to phospho-Raf-1 within the model, as intermediate variables such as Ras-GTP were not measured; and similarly the role of nuclear phospho-ERK-1/2 in promoting its own dephosphorylation is modeled as a self-interaction, as the induced phosphatase DUSP4 was not measured.The reaction kinetic diagram describing the signaling processes  = 0, fact(EC50) = 0.5 and fact(1) = 1. EC50 is the fractional activation of an input species at which a half-maximal activation of an output species is induced;where B and K are constrained such that fInhibition: Represented as the negative of activation;Dephosphorylation: Represented as the product of inhibition with the fractional activation of the output species, as the abundance of the output species determines the maximal level of dephosphorylation that can occur.Three types of interactions between species were used to construct the mathematical model:In addition, shuttling between the cytoplasmic and nuclear compartments was modeled to be linearly proportional to the fractional activations of the species in the respective compartments. Pathway crosstalk was implemented by summing individual species interactions. Using this approach, the ERK-MAPK signaling network in Fig. 2+, plasma membrane CaM, cytoplasmic phospho-Raf-1, cytoplasmic phospho-MEK-1/2, nuclear phospho-MEK-1/2, cytoplasmic phospho-ERK-1/2, and nuclear phospho-ERK-1/2, respectively . The scaling by r of the shuttling terms was to account for volume effects between the cytoplasm and nucleus. The cytoplasmic-nuclear volume ratio was set to vary linearly from 2 (deep) to 5 , as derived from the immunofluorescence data  [MEKcn = 0.05, sMEKnc = 0.5, sERKcn = 0.01 and sERKnc = 0.01) were selected to reflect experimentally-determined ratios of cytoplasm-to-nucleus and nucleus-to-cytoplasm transport coefficients [et al. [Default parameters were used for the normalized Hill differential equations  . MEK andficients . It is dficients . There m [et al.  are deri [et al. , 70. The2+ and plasma membrane CaM activation levels across the epidermis. The Ca2+profile was modeled based on a previous experimental report [b and a are the baseline levels and amplitudes of the input fractional activations respectively  within the epidermis.The cell-autonomous signaling model (above) is driven by spatial profiles of extracellular Cal report  and CaM b and a. Differential equations were implemented in MATLAB  and solved numerically for each spatial partition using the ode23 function to find the steady-state .A least-squares fit to the aggregate cytoplasmic and nuclear phospho-ERK-1/2 immunofluorescence data was performed with the MATLAB lsqnonlin and polyfit functions to optimize values of


Transmission of human T-lymphotropic viruses (HTLV) occurs from mother to child, by sexual contact and blood transfusion. Presently, in most centres in Nigeria, there is no routine pre-transfusion screening for HTLV. The study aims to determine the prevalence of HTLV-1 and HTLV-2 among healthy blood donors at a tertiary centre in Lagos.A cross-sectional study was carried out at the blood donor clinic of the Lagos State University Teaching Hospital (LASUTH), Ikeja. About 5mls of venous blood was collected from each subject into a sterile plain bottle after obtaining subject's consent. The serum separated and stored at -200C. Sera were assayed for HTLV by an enzyme-linked immunoassay (ELISA) for the determination of antibodies to HTLV 1 and HTLV -2. Western blot confirmatory testing was done on reactive samples. All donors were also screened for HIV, HBsAg and HCV by rapid kits.The seroprevalence of HTLV -1 by ELISA was 1.0% and 0.5% by Western Blot among blood donors. A total of 210 healthy blood donors were enrolled. Only 2 (1.0%) blood donors were repeatedly reactive with ELISA test. On confirmatory testing with Western Blot, 1 (0.5%) blood donor was positive for HTLV. All the healthy blood donors were negative for HIV, HbsAg and HCV. None of the 210 blood donors had been previously transfused; as such no association could be established between transfusion history and HTLV positivity among the blood donors.The seroprevalence of HTLV in this environment is low among healthy blood donors. The human T-lymphotropic viruses, type 1 (HTLV-1) and type 2 (HTLV-2), were the first human retroviruses discovered . Human TInfections of HTLV-1 and HTLV-2 are lifelong with asymptomatic carrier state . Over 20Transmission of HTLV-1 occurs from mother to child , 15, by Sexual transmission of HTLV-1 is bi-directional , 23. HowTransmission of HTLV-1 by blood transfusion occurs with transfusion of cellular blood products  but not with the plasma fraction or plasma derivatives from HTLV-1-infected blood . SeroconAs HTLVs are transmitted through blood transfusion, screening for antibodies and discarding seropositive units should efficiently interrupt this transmission. Concern about HTLV-1 transmission through blood transfusion has led to the introduction of routine blood-donor screening for antibodies to HTLV-1 in developed countries , 28. ThePresently at the Lagos State University Teaching Hospital (LASUTH), as in most other centres in Nigeria, there is no routine pre-transfusion screening for HTLV-1. The study aims to determine the prevalence of HTLV-1and HTLV- 2 among healthy blood donors at a tertiary centre in Lagos.A cross-sectional study was carried out at the blood donor clinic of the Lagos State University Teaching Hospital (LASUTH), Ikeja, between February and May 2012.Ethical clearance was obtained from the Health Research and Ethics Committee of LASUTH.Inclusion criteria: All fit blood donors who gave informed consent during the period of the study.Exclusion criteria: Individuals with risk factors for Transfusion Transmissible Infections (TTI). 2. Individuals who were positive for HIV and/or hepatitis B on rapid screening. 3. Donors who did not give consent.Sample size was determined using the Yamane (Sloven)  formula:N = N/ (1 + N (e) 2)Where: n = minimum sample size required; N = the population size; e = degree of accuracy desired i.e. confidence interval, expressed as a decimal (±5% = ±0.05)From LASUTH Blood Bank, records of average number of healthy blood donors seen over a three month period;N = 400 n =400/ (1 + 400(0.05)2) =400/ (1 + 400(0.0025)) = 400/1 + 1 = 400/2 n= 200; Minimum sample size = 200 healthy blood donors.About 5mls of venous blood was collected from each subject into a sterile plain bottle after obtaining subject's consent. The blood was allowed to clot completely before centrifugation. The serum was separated within 6 hours and stored in sterile cryovials at -200C.Serum samples were assayed for HTLV using the HTLV 1 and 2 Ab versions ULTRA by Diagnostic Bioprobes Srl, Italy, an enzyme-linked immunoassay for the determination of antibodies to HTLV 1 and HTLV-2 in serum and plasma. Western blot confirmatory testing (MP Diagnostics (MPD) HTLV Blot 24 Singapore) was done on reactive samples.The sera of donors were also tested for the presence of antibodies to the human immunodeficiency virus (HIV) with Alere Determine TM HIV-1/2 test kit, ( Belgium) hepatitis B surface antigen (HbsAg) with Micropoint HbsAg Gold TM rapid screen test  and hepatitis C virus with DiaSpot HCV test kit . They are rapid screening test kits for the qualitative detection of the respective antibodies in whole blood, plasma or serum.The data was recorded in compatible computer and analysed withEpi-Info version 3.5.3 software. The mean, median, standard deviation and other parameters of statistical location were generated as necessary for continuous data. Tests of statistical significance between variables included chi-square analysis and Fischer's exact for discrete data. Level of significance was set at p < 0.05.The seroprevalence of HTLV -1 by ELISA was 1.0% and 0.5% by Western Blot among blood donors. None of the 210 blood donors had been previously transfused; as such no association can be established between transfusion history and HTLV-1 positivity among the blood donors.A total of 210 healthy blood donors were enrolled. Of the healthy blood donors, 184 (87.6%) were males and 26 (12.4%) were females . The meaAll the 93 single blood donors were HTLV-1 negative. Of the 117 married blood donors, 1 (0.9%) was HTLV-1 positive and 116 (99.1%) were HTLV-1 negative. There was no association between marital status and HTLV-1 positivity. P = 0.324 Of the 210 blood donors, only 1 (0.5%) was HTLV-1 positive and was aged 23 years (age range 20 -29 years). P = 0.614. Among the 184 male healthy blood donors, only 1(0.5%) was HTLV-1 positive. All 26 females were HTLV-1 negative. P = 1.000. Only 2 (1.0%) blood donors were repeatedly reactive with ELISA test. On confirmatory testing with Western Blot, 1 (0.5%) blood donor was positive for HTLV-1. All the The seroprevalence of HTLV-1 of 0.5% among healthy blood donors in this study is similar to the study by Analo et al in 1998 in Lagos in which 0.7% of blood donors were HTLV-1 positive . It is lThe prevalence of HTLV-1 among blood donors in this study is however much higher than that among blood donors in North America and Europe, where seroprevalence is very low, for example, 0.01-0.03% in USA and Canada , 34, 0.0History of previous blood transfusion appeared to have no impact on HTLV-1 status in this study. None of the blood donors had been previously transfused. This is at variance to studies that reported past history of transfusion as an important risk factor for HTLV-1 seropositivity , SeveralThis is not in conformity with studies that have shown the frequency of HTLV-1 transmission to be much higher from male to female than from female to male , 24. ThiThe seroprevalence of HTLV-1 in this environment is low among healthy blood donors.


Previelations . Althougelations . An impoelations .960 HITs were uploaded to mTurk and 121 naïve subjects participated. Based on data quality, 91% of HITs were retained at a cost of $0.132 AUD per HIT. The mTurk data was compared to two supervised lab datasets. Lab and mTurk performance functions were very similar Figure  and high


Integration of the provirus into the host genome is an essential step in the establishment of a chronic retroviral infection. Retroviruses do not integrate at random. Each genus of retrovirus preferentially integrates in genomic sites associated with certain genomic features. This preference is thought to result from an interaction of the pre-integration complex with a specific host co-factor. Such an interaction has been well documented between HIV-1 integrase and lens epithelium-derived growth factor (LEDGF) and between MLV and members of the Bromoextraterminal Domain Protein (BET) family. HTLV-1 also has a reproducible preference to integrate near various genomic features; however, the putative host co-factor of HTLV-1 integrase remains unknown. We present evidence that demonstrates that the HTLV-1 integrase co-factor may be the protein phosphatase 2A (PP2A) complex. Two independent protein interaction screens (yeast-2-hybrid and co-immunoprecipitation) both revealed an interaction between HTLV-1 integrase and the PP2A holoenzyme. We used co-culture with HTLV-1 producer cell line MT2 to infect rat cell lines containing an inducible knockdown of PP2A. We found that PP2A-knockdown altered the integration profile of these de novo integration sites. In contrast to infection in PP2A-competent cells, integration sites in PP2A-knockdown cells lacked the characteristic proximity to genomic features associated with transcriptional activity  and to epigenetic marks associated with active chromatin. We are in the process of investigating how PP2A inhibitors and shRNAs targeting specific regulatory subunits of the PP2A holoenzyme influence integration site selection.


High HTLV-1 proviral load (PVL) is mainly found in infected individuals with HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). However one third of asymptomatic carriers may have high PVL.This study aimed to evaluate the impact of PVL in the activation of T lymphocytes of asymptomatic individuals infected with HTLV-1.+, CD28+, CD45RO+, CD69+, CD62L+, HLA-DR+), FoxP3+ and intracellular IFN-γ expression were evaluated on both CD4+ and CD8+ T-lymphocytes from asymptomatic carriers with PVL ≥ and < 1% of infected cells, using flow cytometry. HTLV-1 proviral load was determined using real-time PCR.Membrane activation markers , CD4+HLA-DR+  and CD4+IFN-γ+  T-cells, than healthy donors. HTLV-1 PVL was directly correlated with the proportion of CD4+CD25+CD45RO+ T-cells . Moreover, a significant increase in the proportion of CD4 + FoxP3+ T-cells was observed in HTLV-1-infected individuals, compared to healthy donors.Asymptomatic carriers with PVL ≥ 1% presented a higher frequency of CD4+ and CD8+ T-lymphocytes in asymptomatic individuals. Prospective studies should be conducted to evaluate whether asymptomatic individuals with higher PVL and high immune activation are more prone to developing HTLV-1-associated diseases.HTLV-1 PVL is associated with activation of both CD4The online version of this article (doi:10.1186/1471-2334-14-453) contains supplementary material, which is available to authorized users. The human T-lymphotropic virus type 1 (HTLV-1) was the first retrovirus known to cause disease in humans, initially isolated from the lymphocytes of a patient with cutaneous T-cell lymphoma in 1980 . This viThe association with these infectious diseases suggests that HTLV-1 may impair the host immune response, possibly leading to immunosuppression. Similar to the human immunodeficiency virus (HIV), HTLV-1 also integrates its genome into host cells, thereby establishing a persistent chronic infection. While HIV induces a potent immunodepression by destroying CD4+ T-cells, HTLV-1 promotes the spontaneous proliferation of CD4+ and CD8 + T-cell subsets, as well as NK cells –13.in vitrothymidine . Incorporated [3H]thymidine was measured in terms of mean counts per minute (CPM) using β-radiation counter . Spontaneous proliferation of PBMCs was considered when a mean CPM value ≥ 500 due in triplicates  was obtained.Blood samples were collected from all study subjects in heparin tubes and peripheral blood mononuclear cells (PBMC) were isolated using a Ficoll-Paque Plus density gradient centrifuge . Cultures of 105 events were analyzed per sample.50 μl of whole blood was incubated for 15 min at room temperature with anti-CD4  and the following monoclonal antibodies: CD45RO, CD25, CD69, CD62L, CD28, and HLA-DR . Erythrocytes were subsequently lysed with FACS™ lysing solution . After a final wash, the cells were fixed in PBS containing 1% paraformaldehyde. Alternatively, for detection of intracellular FoxP3, 80 μl of whole blood was incubated for 15 min at room temperature with anti-CD3 and CD4 monoclonal antibodies , after 15 min the cells were washed twice with PBS-BSA-Saponin 0.2% and blocked with AB serum 1% for 5 min. After that, the cells were incubated with anti-FoxP3 for 30 min. The cells were washed and fixed. Analyses were performed using FACSaria II  and the software FlowJo 7.5 . At least 105 cells/well) were cultured in RPMI 1640  medium, supplemented with 25 mM of HEPES, 2 mM of L-glutamine, 1 mM nonessential amino acids, 1 mM sodium pyruvate, 100 U/mL of penicillin, 100 g/mL of streptomycin and 10% pooled human AB serum (Sigma), and plated in triplicate in 96-well U-bottom plates , then incubated for 4 h at 37°C under 5% CO2. Next, monensin and brefeldin (3 μg/ml) in AB serum (10%) (Sigma) were added to each culture, and then reincubated for 16 h. The cells were then washed with PBS-BSA-PFA-azide (2 mL) and stained with monoclonal anti-CD4, and antiCD3 antibodies for 15 min at 4°C. The cells were then fixed in PBS containing 4% paraformaldehyde 200 μl for 20 min at room temperature, followed by two washing cycles with 0.1% PBS-BSA-Saponin. Positive control cells were cultured with 2 μg/ml phytohaemagglutinin (PHA) (Sigma). To conduct intracellular cytokine staining, the cells were then reincubated for 30 min with anti-IFN-γ-PE monoclonal antibodies or isotype controls, then washed with 0.1% PBS-BSA-Saponin, and washed again with PBS-BSA-PFA-azide. Cells were analyzed using a FACSort flow cytometer and data was interpreted by Cellquest software .PBMCs  was attached at 4,829–4,858 bp of the HTLV- 1 reference sequence (HTLVATK). Albumin DNA was used as an endogenous reference and HTLV-1 PVL was calculated as the ratio of [(HTLV-1 DNA average copy number)/] × 2 × 106 and expressed as the number of HTLV-1 copies per 106 PBMCs.PBMCs were obtained from EDTA blood using density gradient centrifugation and cryopreserved until use. DNA was extracted using a DNA extraction system . HTLV-1 proviral load was quantified using a real-time TaqMan polymerase chain reaction (PCR) method, as previously described . BrieflyData are expressed as median and interquartile range (25th percentile and the 75th percentile). Kruskal–Wallis non-parametric analysis of variance with the Bonferroni-Dunn multiple comparison tests was used to compare healthy donors, asymptomatic with PVL ≥ 1% and < 1% of infected cells groups. The Fisher exact chi-square test was used to compare lymphoproliferation frequencies. The correlations were performed by Spearman correlation test. Significant differences were considered for p <0.05. GraphPad Prism 5  Software was used for all statistical analyses.+ T-lymphocytes between PVL ≥ 1% and <1% HTLV-1-infected groups .The median PVL in all HTLV-1-infected carriers was 1.5% of infected cells , 60% of them had PVL higher than 1% of infected cells. There were no statistically significant differences in markers of cellular activation of CD4+CD25+CD45RO+ T-cell subset was directly correlated to the HTLV-1 PVL .Moreover, the frequency of CD4+ T-cell subset  compared to healthy donors  (p = 0.01).Concerning the CD8t Figure B, the fr+IFN-γ+ T-cells (4.5%) was observed in the PVL ≥ 1% group, compared to healthy donors (1%) (P = 0.01), while frequencies of CD8+IFN-γ+ T-cells was similar for HTLV-1 infected groups and healthy donors   was observed in the group infected with HTLV-1, compared to healthy donors. These results were similar to those obtained by other studies [+ cells and its expression does not necessarily convey regulatory function [On the other hand, a fivefold increase in the frequency of regulatory T cells (CD4 studies , 39 indifunction , 41. In + and CD8+ T-lymphocytes in asymptomatic individuals. Prospective studies should be conducted to evaluate whether asymptomatic individuals with higher PVL and high immune activation are more prone to developing HTLV-1-associated diseases. The HTLV-1 PVL may be a relevant marker in monitoring asymptomatic individuals.In summary, the results presented herein indicate that intermediate HTLV-1 PVL is associated with activation of both CD4


A protein kinase called the mechanistic target of rapamycin (mTOR) is a well-known regulator of cellular growth. In fact, several studies have indicated that the kinase activity of mTOR is required for mechanically-induced increases in skeletal muscle protein synthesis and hypertrophy. Previous studies have also determined that the lipid messenger phosphatidic acid (PA) plays a critical role in the stimulation of mTOR signaling and, an increase in PA concentration is sufficient for the activation of mTOR signaling. However, the mechanism by which PA stimulates mTOR is currently unknown. A primary target of mTOR includes the phosphorylation of p70 on the threonine 389 residue (P-p70-389), and thus, is a commonly accepted readout for the activation of mTOR. PA can be synthesized from a variety of reactions via multiple reactants. Therefore, the purpose of this study was to compare the effects of various PA precursors on their ability to stimulate mTOR signaling and determine if any other phospholipid species are also capable of stimulating mTOR signaling.2C12 myoblasts were plated at approximately 30% confluence and grown for 24 hours in 10% FBS High Glucose DMEM. Cells were switched to 2mL/well serum free high glucose DMEM (no antibiotics) for 16 hours prior to the experiment. Cells were approximately 70% confluent at the time of the experiment. Cells were then stimulated for 20 minutes with vehicle (Control) or 10, 30 or 100µM of soy-derived phosphatidylserine , phosphatidylinositol (S-PI), phosphatidylethanolamine (S-PE), phosphatidylcholine (S-PC), PA , lysophosphatidic acid (S-LPA), diacylglycerol (DAG), glycerol-3-phosphate (G3P), and egg-derived PA (E-PA). Cells were harvested in lysis buffer and subjected to immunoblotting. The ratio of P-p70-389 to total p70 was used as readout for mTOR signaling.CS-PI, S-PE, S-PC, DAG, and G3P elicited no increase in the ratio of P-p70-389 to total p70 compared to vehicle stimulated cells. In contrast, elevated mTOR signaling was observed at all tested concentrations of S-PS , S-LPA , and S-PA . Egg-PA induced an 873% increase in mTOR signaling with the 100µM dose (P<0.05), whereas no significant increase was observed with the 10 or 30µM doses.S-PA, S-LPA and S-PS are each sufficient to induce an increase in mTOR signaling. Therefore, they may be capable of enhancing the anabolic effects of resistance training and contributing to muscle accretion over time. Furthermore, S-PA is a more potent stimulator of mTOR signaling than PA derived from egg.


LRRC10 mutations in human idiopathic dilated cardiomyopathy (DCM) and Lrrc10 homozygous knockout mice develop DCM, strongly linking LRRC10 to the molecular etiology of DCM. LRRC10 localizes to the dyad region in cardiomyocytes where it can interact with actin and α-actinin at the Z-disc and associate with T-tubule components. Indeed, this region is becoming increasingly recognized as a signaling center in cardiomyocytes, not only for calcium cycling, excitation-contraction coupling, and calcium-sensitive hypertrophic signaling, but also as a nodal signaling hub where the myocyte can sense and respond to mechanical stress. Disruption of a wide range of critical structural and signaling molecules in cardiomyocytes confers susceptibility to cardiomyopathies in addition to the more classically studied mutations in sarcomeric proteins. However, the molecular mechanisms underlying DCM remain unclear. Here, we review what is known about the cardiomyocyte functions of LRRC10, lessons learned about LRRC10 and DCM from the Lrrc10 knockout mouse model, and discuss ongoing efforts to elucidate molecular mechanisms whereby mutation or absence of LRRC10 mediates cardiac disease.Leucine-rich repeat containing protein 10 (LRRC10) is a cardiomyocyte-specific member of the Leucine-rich repeat containing (LRRC) protein superfamily with critical roles in cardiac function and disease pathogenesis. Recent studies have identified  Dilated cardiomyopathy (DCM) and hypertrophic cardiomyopathy (HCM) are the most common primary myocardial diseases with a prevalence of at least 1 in 2500 and 1 in 500 individuals, respectively,  was identified based on its cardiac-specific expression pattern  mice  and Akt signaling  15.5, prior to the development of cardiac dysfunction at E17.5 or eccentric ventricular remodeling that occurs in adulthood, revealed upregulation of the actin cytoskeleton and focal adhesion gene pathways to be the most significantly dysregulated gene networks , and parvin-α, which are all induced in the heart in response to biomechanical stress  is regulated by LRRC26, which serves as an auxiliary γ-subunit to regulate BK channel activity  channel  or human pluripotent stem cells  Grant HL-067050 and American Heart Association (AHA) Grant 12GRNT12070021 (to YL), AHA Predoctoral Fellowship 11PRE5580012 (to MB), and NIH Molecular and Environmental Toxicology Training Grant T32ES007015.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.


The melanoacanthoma is a rare benign pigmented tumor, characterized by a fast radial growth and clinical behavior similar to melanoma. Color changes in oral mucosa and dermis are consequence of increased melanocyte activity as response to an irritant factor. There is a vast phenotypic variety. It is difficult to distinguish between a benign pigmented lesion and a melanoma at its early stage. Due to its clinical relevance is crucial to diagnose possible malignancy of the lesions.The aim of this article is to conduct a systematic review of all published articles, as well as update and evaluate etiologic factors and clinicopathological features.We carried out a search in the Medline database (PubMed) using the key words “oral melanoacanthoma” AND “oral melanoacanthosis” AND “oral melanoepithelioma”. Inclusion criteria were all published articles since its discovery. Demographic data, histological features and immunohistochemical findings were extracted from the full articles.A total of 56 articles were analysed. 114 injuries drawn from these articles were studied, a total of 115 injuries with our contribution case. The 74.78% of authors claim a reactive pathogenesis. The average age of lesión appearance is 34.79 years, with an age range of 5-87 years. There is a predominance of the female sex in solitary phenotype 3: 2 and a ratio of women to men 5: 3 if it is multifocal phenotype. Bilateral phenotype is slight higher in women of 2: 1.Histopathological analysis of the lesión is vital to diagnose malignancy. Therefore, any heterogeneous, pigmented lesion with irregular edges, raised surface, fast growth and abrupt appearance should be biopsied. More emphasis on the potential irritants should also be put to improve the quality of life of our patients and to reduce morbidity of melanoacanthoma, as well as, several similar clinical behavior disease. Key words:Melanoacanthoma, oral cáncer, diagnosis. Cutaneous variant occurs mainly in fair-skinned adults while the oral melanoacanthoma has a predilection for blacks and younger patients .Radial growth it is a high potential pathognomonic sign, it can mask a subyacent melanoma .These characteristics have been studied with electronic microscopy, and several tests had been used like: inmunoprecipitation test with the aim of analize patient´s serum to search antibody antimelanoma, inmunofluorescence to look for present anthygens on melanoma lesion: inmunohistochemical studies have demostrated melanocitic reactivity of the melanocytes which reside on the basal, parabasal and cellular espinous strate for the marquer HMB-45; the protein S-100 serves as marquer of the presence of melanocitics dendritics cells, very useful for its diagnosis confirmation, as the marquer Melanin- A also is used with this purpose .Quirurgical exéresis shows a great ratio of success without recurrences Fig. . It offeThe aim of the present article is to make a systematic review of all the published cases, as to actualize and evaluate ethiologic factors and it clinicopathologic characteristics.A systematic, computerized database search was conducted using the National Center for Biotechnology Information (NCBI) to search MEDLINE (Pubmed). The search was conducted using the following MeSHterms:” “oral melanoacanthoma” AND “oral melanoacanthosis” AND “oral melanoepithelioma”.For the initial selection, we selected all articles published since melanoacanthoma. Demographic data, histological characterestics and immunohistochemical findings were taken from the full text. From the literature a total of 59 articles, in relation with mela-noacanthoma, were obtained, three of those were exclude after complete reading. We evaluated 56 articles. A total of 115 patients, including the case presented by our team, were diagnosed.Figure The review of the literature shows that oral melanoacanthoma affects patients aged between 5 and 87 years, with a mean age of 34.79 years. There is higher prevalence in women, 54.4% versus 38.4% in men.The ratio female-male is 3: 2. There is a predominance of the female sex in solitary phenotype 3: 2 and when the multifocal phenotype is the ratio female-male is 5: 3. In the bilateral phenotype is slight higher in women, 2: 1.The solitary phenotype appears more frequently (18.26% of cases) than the multifocal phenotype (13.91% of cases).The locations from highest to lowest frequency are buccal mucosa 33.9%, 13.04% palate, 5.22% lips, 13.91% alveolar mucosa including retromolar área and lingual mucosa, 3.48% tongue, 4.35% back, 3.48% abdomen, 3.48% scapula, 3.48% ear, 3.48% eyelid, 2.6% leg, 2.6% buttock, 1.74% neck, 1.74% floors mouth, 1.74% nose, 1.74% armpit, chest 0.87%, 0.87% vermilion lipstick, hip 0.87%, 0.87% base of the penis, arms 0.87% 0.87% temporal region, 0.87% submental region, 0.87% scrotum groin area, 0.87% preauricular area ,0.87% shoulder and 0.87% forehead.The most frequent presentation is blackish brown in 40.8% of cases, followed by bluish black 3.2%, 1.6% reddish brown and grey 0.8%.It has a predilection for black race (37.39%) followed by caucasian (19.13%), latin American (3.47%) and Asians (3.47%). 77.42% of the authors assert that the etiology is related to irritative factors.The variability of phenotypic expression justifies the controversy of the classification of this entity. There are different phenotypes with multiple expressions: cutaneous or oral melanoacanthoma, painful or asymptomatic, fast growing or stable, multiple or solitary melanoacanthomas; as well as different locations and histopathologic features.et al. consider that oral melanoacanthoma is a tumor wrongly named and defined it as an unusual proliferation of dendritic melanocytes mucositis in the epidermis , a case of simple hydroacanthoma with a malignant transformation into a porocarcinoma. This extremely rare tumor classification is highly complex due to its histopathological similarities with other lesion and degenerative changes of any lesion with age. The relationship between melanocytes and keratinocytes is very similar to the realtion observed in the melanoacanthoma . This raSimon believes for there are three variants of melanocytic seborrheic keratosis: irritant, non-irritating and nested variants. He considers that oral melanoacanthoma is an irritant seborrheic keratosis and he suggests the term melanoacanthoma should be removed .Concerning relations between melanoacanthoma and seborrheic keratosis it should be noted that the only difference between the two processes is that in melanoacanthoma can be found many melanocytes at every level of tumor epidermis, while in seborrheic keratoses, melanocytes are not increased and they can only be found in basal area . AuthorsClinical appearance similar to other pigmented lesions, family history, drug use or systemic drugs, hormones, heavy metals and changes in the morphological pattern are important for the differential diagnosis . Some paBack to melanoacanthoma histology, this is a pigmented tumor that exhibits great morphological variability. Pathological characteristics show stratified squamous epithelium with proliferation of melanocytes and melanin presence in the basal layer and suprabasal layers without invading the underlying connective tissue, prone to the central keratinization (endoqueratinización). The predominant cell pattern prickly keratinocytes or basal differentiation is present in different areas. Melanocytes have extensive dendritic processes and striking areas of acanthosis .The presence of inflammatory infiltrate is found in the great majority of melanoacanthomas. The block in the transference of melanin from melanocytes to keratinocytes is the nature of this entity. Alteration in the normal pattern and speed differentiation of keratinocytes alters cell characteristic of keratinocytes surface which inhibits pigment donation.The etiological hypotheses of reactive origin pigmented lesion is supported by the frequent presence of inflammatory infiltrate of lymphocytes.Langerhans cells are present in every Malpighian layer except from the basal layer. These cells are related to proliferation control of keratinocytes. Therefore, the study of Langerhans cells is interesting due to its disposition at every layers .The histopathological analysis of the melanoacanthoma suspicious lesion is crucial to rule out malignancy, as it may hide a subyacent oral melanoma. Any heterogenous pigmented lesion with irregular borders, raised surface, fast growth and sharp appearance should be biopsied. Since over 75% of reported cases indicate an irritating background, more emphasis must be put on the control of them.This can improve the quality of life of our patients and reduce morbidity of numerous pathologies.


Hepatozoon is an apicomplexan parasite of carnivores transmitted by the tick Rhipicephalus sanguineus trough ingestion. Infections in red foxes (Vulpes vulpes) have been recorded in several parts of the world, including European countries, with variable prevalence values.Hepatozoon spp. in red foxes originating from two counties (Arad and Timiş) of Western Romania.The study aimed to investigate the occurrence of Hepatozoon spp.Fresh/coagulated blood samples from 28 red foxes, killed in sanitary hunting during the routine rabies monitoring, were molecularly analyzed by a conventional PCR using the primers Hep F and Hep R for the presence of Overall, 11/28 (39.2%) investigated samples contained the DNA of the parasite. Most of the positive samples were from Arad County (8/17), while in Timiş County only 3 out of 11 foxes tested were positive.Hepatozoon canis was reported in Romania in dogs but investigations in wildlife were not performed.Occurrence of Rhipicephalus sanguineus) in the area, documented by previous surveys.The relatively high prevalence value of the infection recorded in the current survey can be related to the common occurrence of the brown dog tick (The high prevalence of the parasite among red foxes confirms the presence of sylvatic cycle, highlights the role of wildlife as reservoirs and potential vectors for the infection in the screened region and also the emerging of this pathogen agent in the country.Hepatozoon spp. infection in red foxes from Romania. The findings support the global changing in vector borne diseases expansion.This is the first report of this study was financially supported by CNCS Grant TE_277_116/2010.