This study evaluated the correlation between HTLV-1 proviral load and CD4+ and CD8+ T cells count, and lymphproliferation assay (LPA).A total of 193 HTLV-1-infected subjects underwent an interview, dermatological examination, initial HTLV-1 proviral load assay, CD4A total of 147 patients had an abnormal skin condition; 116 (79%) of them also had SD-HTLV-1 and 21% had other dermatological diagnoses. The most prevalent SD-HTLV-1 was xerosis/acquired ichthyosis (48%), followed by seborrheic dermatitis (28%). Patients with SD-HTLV-1 were older (51 vs. 47 years), had a higher prevalence of myelopathy/tropical spastic paraparesis (HAM/TSP) (75%), and had an increased first HTLV-1 proviral load and basal LPA compared with patients without SD-HTLV-1. When excluding HAM/TSP patients, the first HTLV-1 proviral load of SD-HTLV-1 individuals remains higher than no SD-HTLV-1 patients.There was a high prevalence of skin disorders (76%) among HTLV-1-infected individuals, regardless of clinical status, and 60% of these diseases are considered skin disease associated with HTLV-1 infection. HTLV-1 infection may increase the risk of developing skin disorders. A total of 193 HTLV-1 infected subjects were studied, including asymptomatic carriers and HAM/TSP patients. Of the subjects, 76% had an abnormal skin condition, with a high prevalence both among HTLV-1 asymptomatic carriers and HAM/TSP patients. The most prevalent SD-HTLV-1 was xerosis/acquired ichthyosis (48%), followed by seborrheic dermatitis (28%). Patients with SD-HTLV-1 were older (51 vs. 47 years), had a higher prevalence of myelopathy/tropical spastic paraparesis (HAM/TSP) (75%) and an increased first HTLV-1 proviral load compared with patients without SD-HTLV-1. When excluding HAM/TSP patients, the first HTLV-1 proviral load of SD-HTLV-1 individuals remains higher than no SD-HTLV-1 patients. Thus, skin diseases are highly prevalent among HTLV-1-infected individuals. Adult T-cell leukemia/lymphoma (ATLL), HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) and infective dermatitis associated with HTLV-1 (IDH) are the main diseases caused by human T-cell lymphotropic virus type 1 (HTLV-1) infection There are an estimated 5 to 10 million HTLV-1 infected individuals worldwide and Brazil is considered a highly endemic area for HTLV-1 infection, with the largest absolute number of HTLV-1 infected individuals, with more than one million people living with this virus + and CD8+T cell count.There is a lack of surrogate markers to assess the infected patients who have a higher risk for HTLV-1 associated skin disorders. Moreover, there are few immunological studies among HTLV-1-infected persons who are simultaneously suffering from HAM/TSP and skin diseases. The aim of this study is to evaluate the prevalence of skin disorders in HTLV-1-infected individuals and to correlate this prevalence with the initial HTLV-1 proviral load, and initial CD4In the last 18 years, a cohort of HTLV-infected subjects has been followed in the HTLV-outpatient clinic at the Institute of Infectious Diseases “Emilio Ribas” (IIER), with the support of nurses, nutritionists and physical therapists. From a total 450 HTLV-1-infected individuals, including asymptomatic carriers and HAM/TSP patients, 193 of them were consecutively evaluated for skin disorders from January 2008 to July 2010 by the same dermatologist, blinded for the clinical status to minimize information bias. Demographical and clinical dates were collected, and dermatological examinations were carried out. The HIV co-infected individuals were excluded, but HCV co-infected subjects were included.Ethics Statement: Written informed consent was obtained from all participants, and the IIER ethical board approved the protocol.+ and CD8+T cell counts, and an initial HTLV-1 proviral load. In accordance with previous studies, the following skin disorders associated with HTLV-1 infection (SD-HTLV-1) were considered: xerosis/acquired ichthyosis, seborrheic dermatitis and infective dermatitis associated with HTLV-1 (IDH) + and CD8+ T-cell subsets, fresh whole blood specimens were collected in EDTA tubes and subjected to flow cytometry , using human monoclonal antibodies anti-CD3, anti-CD4, and anti-CD8, labeled with fluorochrome.The patients underwent laboratory studies, including HTLV-1 serological diagnosis, initial CD4pol gene. The albumin gene served as the internal genomic control, and MT2 cells were used as a positive control. The results are reported as copies/10000 PBMCs, and the detection limit was 10 copies The results of the first HTLV-1 proviral load were available in the database. Quantitative proviral DNA levels were detected by a real-time automated PCR method, using TaqMan probes for the 2 test. HTLV-1 proviral load was log-transformed to obtain a normal distribution. Correlations between HTLV-1 proviral load and SD-HTLV were performed using Spearman's rank correlation. Data are expressed as mean ± standard deviation  or median and interquartile range . Statistical significance was set at a p value<0.05.Data were analyzed using SPSS 17.0 software. The association between independent variables and the outcome was analyzed either by Student's t-test or ANOVA , or by Mann-Whitney test , while the association between categorical variables and the outcome was assessed by the XOne hundred ninety-three HTLV-1-infected subjects  underwent a dermatological exam. Mean age of patients was 49.4±12.3 years. Female gender had a higher prevalence of HTLV-1 infection (72%). Regarding the presence of neurological involvement, 38% of the patients had a diagnosis of HAM/TSP. The dermatological examination revealed a high prevalence of skin disorders among the HTLV-1-infected patients (76%). Sixty-five individuals (34%) had one dermatological condition, and 42% (n = 81) of the patients had two or more dermatological conditions. Among the 147 patients that had an abnormal skin condition, 79% (n = 116) had a skin disorder associated with HTLV-1 infection (SD-HTLV-1) (xerosis/ichthyosis or seborrheic dermatitis) and 21% (n = 31) had other dermatological diagnoses. The most prevalent skin disorder associated with an HTLV-1 diagnosis was xerosis/ichthyosis (48%), followed by seborrheic dermatitis (28%).+ and CD8+ T cell counts . The initial HTLV-1 proviral load was significantly higher for the group with SD-HTLV-1 as compared with that for the group without SD-HTLV-1 and asymptomatic for neurological symptoms (p = 0.021).Notably, 76% of the HTLV-1-infected asymptomatic carriers and 88% of the HAM/TSP patients showed some skin disorder in our study. These findings are similar to those described in two previous studies involving asymptomatic carriers and HAM/TSP subjects Excluding HAM/TSP cases, subjects with a diagnosis of SD-HTLV-1 are older than groups with other types of skin disorders or individuals with a normal dermatological exam. However, no significant association was observed with gender. These findings suggest that older HTLV-1-infected individuals, who probably have a longer duration of their viral infection have had more time to develop SD-HTLV-1.Although IDH is the only skin disease in which HTLV-1 infection is a criterion for diagnosis, other skin disorders could also be associated with HTLV-1 infection, including xerosis/acquired ichthyosis and seborrheic dermatitis, as previously demonstrated Staphylococcus aureus and beta-haemolytic streptococci, what additionally leads to chronic antigenic stimulation and persistent inflammation of the skin. Genetic, host and environmental factors have been shown to be associated The exact pathogenic mechanism of IDH still needs to be made clear, but the current view is that a diagnosis of HTLV-1 infection is necessary, leading in susceptible individuals to immune deregulation, with subsequent immunosuppression and superinfection with Xerosis and acquired ichthyosis have been described as the main dermatological manifestations associated with HAM/TSP patients Acquired ichthyosis is a consequence of hypohydrosis that may be secondary to the involvement of the autonomic nervous system, affecting directly the HTLV-1-infected skin cells HTLV-1 proviral load is a laboratorial risk marker for the development of HAM/TSP and other diseases related to HTLV-1 infection 10. Because of these findings, several authors believe that HTLV-1 can modify the function of infected cells, resulting in skin disorders that are caused directly by the presence of the virus in the infected cells As previously shown, HTLV-1 was identified by PCR on skin cells in addition to lymphocytes in HTLV-1-infected persons, regardless of their clinical status Finally, for the first time, a high prevalence of skin disorders (76%) independent of clinical status was disclosed among HTLV-1-infected individuals. These findings may have important implications in the clinical setting in places where this infection is endemic and dermatologists, infectious diseases specialists and general clinicians should be aware of skin presentations of the HTLV-1 infection. Moreover, the influences of demography and co-morbid conditions may be relevant, but have not been fully studied. Thus, data derived from a cohort of referred patients followed in a specialized HTLV clinic may not be a representative sample of the whole population of HTLV-1 patients and therefore unlikely to reflect the prevalence of skin conditions among all HTLV-1 patients.Checklist S1STROBE checklist.(DOCX)Click here for additional data file.


In resource-poor areas, infectious diseases may be important causes of morbidity among individuals infected with the Human T-Lymphotropic Virus type 1 (HTLV-1). We report the clinical associations of HTLV-1 infection among socially disadvantaged Indigenous adults in central Australia.st January 2000 to 31st December 2010 were obtained from the Alice Springs Hospital pathology database. Infections, comorbid conditions and HTLV-1 related diseases were identified using ICD-10 AM discharge morbidity codes. Relevant pathology and imaging results were reviewed. Disease associations, admission rates and risk factors for death were compared according to HTLV-1 serostatus. HTLV-1 western blots were positive for 531 (33.3%) of 1595 Indigenous adults tested. Clinical associations of HTLV-1 infection included bronchiectasis , blood stream infections (BSI) with enteric organisms  and admission with strongyloidiasis . After adjusting for covariates, HTLV-1 infection remained associated with increased numbers of BSI episodes  and increased admission numbers with strongyloidiasis  and respiratory conditions including asthma , lower respiratory tract infections  and bronchiectasis . Two patients were admitted with adult T-cell Leukemia/Lymphoma, four with probable HTLV-1 associated myelopathy and another with infective dermatitis. Independent predictors of mortality included BSI with enteric organisms  and bronchiectasis .HTLV-1 serological results for Indigenous adults admitted 1HTLV-1 infection contributes to morbidity among socially disadvantaged Indigenous adults in central Australia. This is largely due to an increased risk of other infections and respiratory disease. The spectrum of HTLV-1 related diseases may vary according to the social circumstances of the affected population. The Human T-Lymphotropic Virus type 1 (HTLV-1) infects at least 5–10 million people worldwide. In developed countries, the most frequently reported HTLV-1 associated diseases include a fatal hematological malignancy, Adult T-cell Leukemia/Lymphoma (ATLL), and the neurological disorder, HTLV-1 associated myelopathy (HAM), which arise in <10% of HTLV-1 carriers during their lifetime. However, most HTLV-1 carriers live in resource-poor areas where infectious diseases, such as strongyloidiasis, could be more important causes of morbidity. Demonstrating such an effect is difficult due to the resource constraints experienced by developing countries in which populations with a substantial burden of infectious diseases reside in areas that are highly endemic for HTLV-1. This is not the case in HTLV-1 endemic central Australia where Indigenous Australians have, for example, among the highest reported blood stream infection rates worldwide in a setting in which sophisticated medical facilities are readily available. We report that bronchiectasis, blood stream infections and admissions with lower respiratory tract infections and strongyloidiasis are associated with HTLV-1 infection. These conditions were far more common than HTLV-1 associated malignancies or neurological conditions in this socially disadvantaged Indigenous population. The spectrum of HTLV-1 related diseases therefore varies according to the social circumstances of the affected population. The Human T Lymphotropic Virus type 1 (HTLV-1) is an oncogenic retrovirus that preferentially infects CD4+ T cells Strongyloides stercoralisS.stercoralis, HTLV-1 infection is the major risk factor for complicated strongyloidiasis or ‘hyperinfection’, which is associated with pulmonary involvement S.stercoralis may also reduce the latent period required for the development of ATLL Infectious diseases also contribute to HTLV-1 related morbidity and mortality. Severe scabies 2  morbidity codes relating to non-communicable diseases, possible HTLV-1 related conditions and infectious diseases were also extracted (All adults (age ≥15 years) admitted to ASH between 1xtracted . Discharxtracted . HTLV-1 Place of residence was categorized as i) remote (>80 km from Alice Springs), ii) Alice Springs town camp and iii) urban . Remote residence was further divided into quadrants  relative to Alice Springs.S.stercoralis serology, HBV serology and blood cultures were obtained from the ASH pathology data-base. During the study period, S.stercoralis serology was performed using an in-house enzyme-linked immunosorbent assay based on antigen extracts of Strongyloides ratti, which is highly sensitive and specific. A blood culture from which a pathogen was isolated defined a ‘BSI episode’. Repeated culture of the same organism from blood culture was regarded as a separate ‘episode’ only if blood samples were drawn more than one month apart. Blood stream infections excluded potential contaminants including coagulase negative staphylococci, bacillus spp., coryneforms and viridans streptococci unless grown from more than one BC in a 24 hour period and Acinetobacter spp in the absence of an identifiable focus. For statistical analysis, the major BSI pathogens were grouped according to their most likely origin: i) skin (Staphylococcus aureus and Streptococcus pyogenes), ii) respiratory (Streptococcus pneumoniae and Haemophilus influenzae), iii) urinary tract (Escherichia coli) and iv) gastrointestinal tract (Enterobacteriaceae other than E.coli). ‘Definite bronchiectasis’ was defined as an ICD-10 AM code for bronchiectasis that was confirmed by High Resolution Computed Tomography (HRCT) chest according to American College of Chest Physicians criteria. ‘Possible bronchiectasis’ was defined as an ICD-10 AM code for bronchiectasis in the absence of HRCT chest confirmation of this diagnosis. A diagnosis of ATLL Results for Initial screening tests were performed using the Serodia HTLV-1 particle agglutination assay  or Architect rHTLV-I/II assay at the Royal Darwin Hospital, Northern Territory of Australia, (1458) or the Institut Pasteur, Paris (156). Positive samples were again tested using both the Serodia HTLV-1 particle agglutination assay and Murex HTLV-I+II test kit  or an indirect immunofluorescence assay (IFA) using an HTLV-1-transformed human T cell line (MT2)(Institut Pasteur). HTLV-1 serostatus was then confirmed by Western blot  using stringent criteria for all samples for which screening tests were positive.Categorical variables were summarized using frequency and percentage and compared using a Chi-square test or, in the case of small numbers, a Fisher's Exact test. Multiple simultaneous comparisons were adjusted for using a Bonferroni correction. Continuous variables were assessed for significant departures from normality with normally distributed variables summarized using mean and standard deviation (SD) and compared using a t-test whilst skewed variables were summarized using median and inter-quartile range (IQR) and compared using a Wilcoxon rank-sum test.Predictors of HTLV-1 seropositivity were examined using Poisson regression with robust standard errors. Strongyloides admissions (identified by ICD-10 AM codes), rather than serology, were included in the multivariable model because these are more likely to reflect symptomatic strongyloidiasis Incidence rates of admission count by diagnostic group were expressed as a proportion of the total number of HTLV-1 seropositive and seronegative patients respectively. Predictors of admission counts for a range of diagnostic groups according to HTLV-1 seropositivity were examined using negative binomial regression and are presented with their negative binomial 95% confidence intervals. Negative binomial modeling was preferred over straight Poisson regression due to over-dispersion in admission count outcome variables. The model coefficients represent the estimated change in admission counts for a particular level of a predictor variable. The influence of HTLV-1 seropositivity on admission count was adjusted for demography and comorbidities. In the case of admissions with asthma, LRTI, pneumonia and chronic obstructive pulmonary disease, the model was adjusted for both definite and possible bronchiectasis and tobacco smoking. A link test was used to assess the model for specification error whilst overall goodness of fit was assessed using both visual examination of residuals coupled with a likelihood-ratio test and a Pearson goodness-of-fit test.Predictors of hepatitis B surface antigen (HBsAg) positivity were analysed using logistic regression. In this case, a logistic approach was preferred secondary to the rarity of the outcome. Overall model fit was assessed using a Hosmer & Lemeshow goodness-of-fit test.Predictors of time to mortality were examined using Cox Proportional Hazards Regression. Analysis of scaled Schoenfeld residuals were used to assess compliance with the proportional hazards assumption. For this analysis patients with possible bronchiectasis were assumed not to have the condition.All reported p-values are two-tailed and for each analysis p<0.05 was considered significant. All analyses were conducted using Stata version 12 .HTLV-1 screening tests were performed for 1614 Indigenous adults and these were positive for 624 (38.7%) cases. Samples from 605 patients were referred for confirmatory Western blot tests. These were indeterminate in 73 cases (4.6%) and confirmed HTLV-1 infection for 531 patients (33.3%). Patients whose western blot results were indeterminate were excluded from further analysis, as were 74 patients  who died prior to 2005. The subsequent analysis therefore included 1451 Indigenous adults  who were admitted 115,919 times  during the study period.HTLV-1 seropositivity rates among males increased significantly with age ; ≥45 years, 135/319 (42.2%); p = 0.008). Rates were otherwise not significantly different between age groups or genders . SeroposA bronchiectasis-related discharge morbidity code was recorded for 170 patients. Bronchiectasis was confirmed radiologically in 142 patients of whom 81 (57.0%) were HTLV-1 seropositive. Radiologically confirmed bronchiectasis was an independent predictor of HTLV-1 infection in a multivariable model  . HTLV-1 Strongyloides serology was performed for 1126 (77.6%) patients of whom 269 (23.9%) were Strongyloides seropositive . AlthougHTLV-1 carriers were more likely to record a discharge morbidity code for scabies  and had The risk of non-hematological malignancies was significantly reduced among HTLV-1 carriers . Six patNearly 70% of patients  recorded at least one discharge code for sepsis with no focus specified during the study period. Although HTLV-1 carriers more often recorded discharge codes for sepsis with no focus specified  and had Among 988 (68.1%) patients tested, 127 (12.9%) were HBsAg positive of whom 16 (12.6%) were also HBeAg positive . The geoAmong 338 deaths that occurred during 5,739 years of follow-up, 120 (23.7%) were HTLV-1 seropositive and 218 (23.1%) were HTLV-1 seronegative. There was no difference between HTLV-1 seropositive and seronegative patients in median age of death ; HTLV-1 seronegative, 53.2 years; IQR, 44.4, 62.5) . DemograE.coli  remained significant predictors of death after multivariable analysis  and non-communicable diseases  and BSI with Enterobacteriaceae other than analysis . Other rlignancy).In a hospitalized cohort of Indigenous Australian adults, we found an HTLV-1 seropositivity rate (33.3%) that was approximately three times the estimated background rate in central Australia (7.2–13.9%) Pulmonary involvement is common among HTLV-1 carriers elsewhere. Radiological abnormalities, for example, have been reported in 50% of Japanese patients with HAM/TSP and 30% of asymptomatic HTLV-1 carriers who were examined by chest X-ray Nevertheless, clinically significant pulmonary disease is an uncommon feature of HTLV-1 infection in developed countries S.stercoralis infection were found among Indigenous adults in an arid region of Australia that would appear otherwise hostile to soil transmitted helminths. The presence of HTLV-1 infected ‘core transmitters’ who carry a high larval burden may be central to the survival of S.stercoralis in this environment and could increase the risk of S.stercoralis infection among other community members. Strongyloidiasis may also contribute to the very high BSI incidence rates that have been reported in central Australia Consistent with the results of other studies Mycobacterium tuberculosisS.stercoralisIn our Indigenous Australian cohort, respiratory and infection-related morbidity were increased among HTLV-1 carriers in the absence of an increased risk of death. However, an effect of HTLV-1 infection on mortality may be obscured by analysis according to HTLV-1 serological status rather than HTLV-1 proviral load, which is closely associated with HTLV-1 related diseases The retrospective nature of this study results in a number of limitations. First, patients with HTLV-1 related diseases were identified from discharge morbidity codes. Attempts were made to identify other patients with these conditions by contacting specialist medical units to which such patients are referred; however, cases may have been missed if these were not coded or referred appropriately. Consequently, our data are likely to under-estimate the actual burden of HTLV-1 related diseases in this population. The risk of bronchiectasis attributable to HTLV-1 infection is also likely to be underestimated because individuals who had not received radiological confirmation of this diagnosis were assumed not to have the condition. Similarly, the effect of HTLV-1 infection on respiratory conditions may be underestimated because ‘possible’ bronchiectasis was included in the final model to account for the increased risk of respiratory infection resulting from structural lung disease. Determining the strength of other possible associations was dependent on the accuracy of discharge coding; however, with the possible exception of strongyloidiasis noted above, this is unlikely to vary according to HTLV-1 serological status. Finally, we assumed that HTLV-1 infection was acquired in most cases prior to the period in which ICD-10 AM codes were collected. The low annual incidence rate of HTLV-1 seroconversion among discordant couples In a setting of overcrowded housing, inadequate health hardware and poor community hygiene Table S1Disease categories and their ICD-10 AM codes recorded 2005–2010. International classification of diseases 10th revision, Australian modification, codes recorded for 1337 Indigenous adults admitted to Alice Springs Hospital, 2005–2010. These codes formed the basis for subsequent analysis according to the categories listed. Abbreviations: COPD, chronic obstructive pulmonary disease; LRTI, lower respiratory tract infection; ICD-10 AM, international classification of diseases 10 Australian Modification.(DOCX)Click here for additional data file.


Drugs targeting the viral enzyme integrase have been in use for several years as part of the treatment regimen for patients with Human Immunodeficiency Virus Type 1 (HIV-1) and similar classes of compounds have been shown to inhibit Human T-Cell Leukemia Virus Type 1 (HTLV-1) integration in vitro. The current study shows that the clinically approved HIV-1 integrase inhibitor, Raltegravir, as well as the more recent diketo acid derivative, MK-2048, are active inhibitors of HTLV-1 infection in vitro. These agents were effective in inhibiting cell-free and cell-to-cell transmission of HTLV-1 in lymphoid and non-lymphoid cells. The drugs also inhibited HTLV-1 immortalization of human peripheral blood mononuclear cells. A novel adaptation of the Alu assay for viral integration was used to show that the drugs inhibit viral integration without affecting reverse transcription. These data support the administration of Raltegravir and other integrase inhibitors as treatments for patients with HTLV-1 associated diseases.


A determinant of HTLV-1 associated myelopathy/tropical spastic paraparesis (HAM/TSP) development is HTLV-1-infected cell burden. Viral proteins Tax and HBZ, encoded by the positive and negative strands of the pX region respectively, play a key role in HTLV-1 persistence. Tax drives CD4+ T-cell clonal expansion but is the immunodominant antigen. Valproate (VPA), an histone deacetylase inhibitor, has been proposed to trigger Tax expression and expose latent HTLV-1 reservoir to immune destruction. We evaluated the impact of VPA treatment on Tax, Gag and HBZ expression in cultured lymphocytes from asymptomatic HTLV-1 carriers and HAM/TSP patients. Around one-fifth of provirus-positive CD4+ T cells spontaneously became Tax-positive. The estimation rose up to two-thirds of Tax-positive infected cells when VPA was added. VPA enhanced Gag p19 protein release. Tax and Gag mRNA levels spontaneously peaked, before a decline concomitant to HBZ mRNA increase. VPA treatment enhanced and prolonged Tax mRNA expression, while it blocked HBZ expression. This is the first ex vivo study on the balance between Tax and HBZ expression (i.e. sense and antisense transcription). Our data suggest that besides modulation of the expression of Tax, another mechanism involving repression of HBZ may determine the outcome of VPA treatment on HTLV-1-infected cell proliferation and survival.


The majority of Human T cell lymphotropic virus type 1 (HTLV-1) infected subjects are considered as HTLV-1 carriers but a high frequency of urinary manifestations of overactive bladder has been documented in these individuals. The aim of this study was to determine if viral and immunological factors that are associated with development of HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) are also observed in HTLV-1 patients with overactive bladder.This was a cross-sectional study comparing immunological responses and proviral load in 3 groups of HTLV-1 infected subjects: HTLV-1 carriers (n=45), HTLV-1 overactive bladder (n=45) by International Continence Society (ICS) criteria and HAM/TSP patients (n=45) according WHO guidelines.We demonstrated that cells from HTLV-1 overactive bladder produce higher spontaneous levels of IFN-γ, TNF-α and IL-17 than carriers and similar levels of TNF-α and IL-17 to HAM/TSP. Proviral load was higher in HTLV-1 overactive bladder and HAM/TSP than carriers and correlated positively with production of proinflammatory cytokines. In contrast to HAM/TSP, HTLV-1 overactive bladder had serum levels of Th1 chemokines similar to carriers and exogenous addition of regulatory cytokines (IL-10 and TGF-β) decreased IFN-γ production in cell cultures from HTLV-1 overactive bladder.We conclude that HTLV-1 overactive bladder patients have some immunological features and proviral load profiles in common with HAM/TSP. However as they are still able to down modulate the inflammatory response and the recruitment of activated T cells to the central nervous system is not enhanced, they present an oligosymptomatic form of HAM/TSP.


Besides horizontal transmission, HTLV-1 is transmitted vertically mainly through prolonged breastfeeding. Maternal transmission via breast milk is the dominant mode of HTLV-1 spread in high endemic areas, and is inked to the presence of HTLV-1 infected cells  in the milk. Infection in young childhood by such mean appears to be a major risk factor for development of ATL in adults.We developed an in vitro model of epithelial barrier (Caco-2 human enterocytic cell line) to assess the mode of passage of HTLV-1 through the digestive tract. Integrity of the barrier was checked by ultrastructural approach, measurement of the trans-epithelial resistance (TER), and diffusion of fluorescently labeled molecules.When enterocytes were co-cultured with HTLV-1-infected lymphocytes, no structural modifications were detected in intercellular tight junctions. Moreover, the functional integrity of the epithelial barrier was maintained since no TER change was detected in the presence of infected lymphocytes, and passage of small fluorescent markers was unaffected. Although enterocytes were not found to be susceptible to HTLV-1 infection, free infectious HTLV-1 virions were detected in the basal compartment, and such a passage was temperature-dependent, suggesting a transcytotic mechanism of passage. When human dendritic cells were added to the basal compartment, they were found to be productively infected by HTLV-1 that had crossed the epithelial barrier.This study provides the first data on the mode of transport of HTLV-1 across a tight epithelial barrier as it may occur during mother-to-child HTLV-1 transmission during breastfeeding.


Several studies have shown that serum uric acid (UA) is associated with left ventricular (LV) hypertrophy. Serum levels of parathyroid hormone (PTH), which has bbe shown to be correlated with UA, is also known to be associated with cardiac hypertrophy; however, whether the association between UA and cardiac hypertrophy is independent of PTH remains unknown.We investigated whether the relationship between serum uric acid (UA) and LV hypertrophy is independent of intact PTH and other calcium-phosphate metabolism-related factors in cardiac patients.In a retrospective study, the association between UA and left ventricular mass index was assessed among 116 male cardiac patients (mean age 65±12 years) who were not taking UA lowering drugs. The median UA value was 5.9 mg/dL. Neither age nor body mass index differed significantly among the UA quartile groups. Patients with higher UA levels were more likely to be taking loop diuretics. UA showed a significant correlation with intact PTH  but not with other calcium-phosphate metabolism-related factors. Linear regression analysis showed that log-transformed UA showed a significant association with left ventricular mass index, and this relationship was found to be significant exclusively in patients who were not taking loop and/or thiazide diuretics. Multivariate logistic regression analysis showed that log-transformed UA was independently associated with LV hypertrophy with an odds ratio of 2.79 .Among cardiac patients, serum UA was associated with LV hypertrophy, and this relationship was, at least in part, independent of intact PTH levels, which showed a significant correlation with UA in the same population. Several previous studies have shown that subjects with higher uric acid (UA) levels more frequently have LV hypertrophy Parathyroid hormone (PTH), a hormone secreted from parathyroid glands, raises plasma calcium levels by increasing absorption, reducing excretion, and promoting release of calcium from bones; in turn, PTH is regulated by the calcium concentration in the plasma. Besides its effect on calcium homeostasis, serum PTH may have an impact on the development of cardiovascular events Nevertheless, to the best of our knowledge it seems that whether the relationship between serum UA and cardiac hypertrophy, when present, is dependent or independent of serum PTH has not been examined. To this end, we have herein investigated, among cardiac patients, whether the association between UA and LV hypertrophy is dependent on serum PTH levels and other calcium-phosphate metabolism-related parameters, including serum fibroblast growth factor-23 (FGF23), which has also been shown to have a link with cardiac hypertrophy The current retrospective study was approved by the Ethics Committee of Osaka Medical College. The study included only subjects who provided written informed consent for whom sufficient information regarding the data analysis was available.Between 2012 January and 2012 December, 138 male cardiac inpatients for whom sufficient clinical and echocardiographic information was available were enrolled in the current study. Among the 138 patients, 21 patients and one patient were taking xanthine oxidase inhibitor  and uricosuric drugs, respectively, and were subsequently excluded. As a result, 116 patients, mean age 65±12 years, were included in the current study.−1.094×(age)−0.287Blood samples were collected in the morning after an overnight fast. Aliquots of serum and plasma were obtained and stored at -80 degrees immediately until use. Ca, IP, C-reactive protein (CRP), and B-type natriuretic peptide (BNP) were measured by routine laboratory methods. When serum albumin was 4 mg/dL or lower, serum Ca levels were corrected by using the formula: Ca+, and designated as corrected Ca (cCa). Serum levels of intact PTH (iPTH) and 25(OH)D were measured using electrochemiluminescence and a competitive protein binding assay . Serum levels of intact FGF23 were measured using a two-step FGF23 enzyme immunoassay (ELISA) kit  according to the manufacturer’s instructions. Measurement of FGF23 was performed by two experienced researchers in the institute: the interclass correlation coefficient (ICC) of intra- and inter-operator reliability for FGF23 was 0.98 and 0.95, respectively. The eGFR was calculated by the following Modification of Diet in Renal Disease equation modified for Japanese subjects: eGFR  =  194×(serum creatinine)3-LVDd3]+0.6 0.425×(height)0.725×0.007184. The LVM index (LVMI) was then calculated as the ratio of LVM to BSA. When the LVMI was greater than 118 g/m2, LV hypertrophy was defined to be present Echocardiographic examinations were performed with a Vivid 7 Dimension equipped with a multi-frequency transducer . Left ventricular end-diastolic dimension (LVDd), interventricular septal thickness (IVST) and posterior wall thickness (PWT) were measured at end diastole. For calculation of the LV mass (LVM), we used the formula proposed by Devereux et al. Patients were divided into the following 4 groups according to their LVMI and the relative wall thickness (RWT), which was calculated as PWT×2/LVDd with a partition value of 0.44 The LV ejection fraction (LVEF) was calculated either by modified Simpson’s method using the apical 4-chamber view, designated LVEF-Simpson, or by the Teichholz’ formula, designated LVEF-Teichholz Baseline characteristics were assessed with standard descriptive statistics. Data were expressed as either mean ± standard deviation or median and interquartile range. A Spearman rank correlation test was used to assess the correlation between two variables. For multivariate analysis, multivariate linear regression and multivariate logistic regression analyses were used to examine the relationship with, respectively, LVMI and LV hypertrophy. Log-transformed FGF23 (log[FGF23]) and intact PTH (log[iPTH]) were confirmed to be normally distributed by the Kolmogorov-Smirnov test. Data analysis was performed by IBM SPSS statistics version 21.0 . A value of P<0.05 was taken to be statistically significant.Among the 138 male patients, the median UA level was 5.9 mg/dL. Neither age nor body mass index differed significantly among the four UA quartile groups . IschemiLaboratory and echocardiographic data were compared across the UA quartile groups by the Kruskal-Wallis test . Serum cBy Spearman’s rank correlation test, serum creatinine, intact PTH, BNP, and LVMI were positively, and eGFR was negatively, associated with serum UA . LVEF-TeMultivariate linear regression model showed that serum UA was significantly associated with LVMI after adjustment for age , model 12), UA, intact PTH, and FGF23 with LV hypertrophy by multivariate logistic regression analysis. We included FGF23 in this model because we previously found a significant association between FGF23 and LVMI in cardiac patients Next, we analyzed the association of age, high blood pressure (systolic blood pressure≥140 mmHg and/or diastolic blood pressure≥90 mmHg), chronic kidney disease . Both UA and intact PTH were associated with LV hypertrophy by univariate analysis. The association between serum UA and LVMI, and that between serum UA and LV hypertrophy were shown to be independent of serum intact PTH and other possible confounders, including age, blood pressure, renal function (eGFR), by linear regression and logistic regression analyses, respectively.To date, several studies have investigated the association between serum UA and LV hypertrophy. It is also important to determine whether the observed association, if present, is dependent or independent of other possible confounding variables because higher serum UA levels may be associated with hypertension, renal dysfunction, diabetes, conditions which are also related to LV hypertrophy Similar to the case of serum UA, the relationship between PTH and LV hypertrophy has been examined in several previous studies. Fujii et al. reported that a relationship between intact PTH and LV hypertrophy in patients with chronic dialysis may be observed only when intact PTH levels are extremely high Several studies have described the association between serum UA and PTH. Serum UA levels were significantly correlated with intact PTH levels among patients with primary hyperparathyroidism In the current study, we showed that serum UA was associated with LV hypertrophy independently of PTH levels and other possible confounders, including age, high blood pressure chronic kidney disease, and FGF23 level, in cardiac patients. In this study population, PTH also showed a positive and independent association with LV mass. The observed relationship observed between UA and LV hypertrophy is not likely to be due to diuretic medications, which can affect serum UA levels, for patients with cardiac hypertrophy and heart failure, because the association between serum UA and LV hypertrophy was observed among patients who were not taking loop and/or thiazide diuretics. Due to the small number of the subjects who were taking aldosterone antagonists, we did not assess whether such drugs would modify the relationship between serum UA and LV hypertrophy. Along with the measurement of renin or aldosterone concentrations, this point should be examined in future studies, as it was recently suggested that activation of mineralocorticoid receptor and secretion of PTH may mutually affected It was found that serum UA was significantly associated with eccentric, but no concentric, LV hypertrophy; however, these assessments should be repeated among a higher number of study patients. We also found that serum UA was not associated with LVEF determined by modified Simpson’s method. Whether patients with hitgher serum UA are more susceptible to LV hypertrophy than systolic dysfunction awaits evaluation in future studies.Several attempts have recently been made to evaluate whether reduction of UA will lead to an improvement in cardiac abnormalities. For example, Rekharaj et al. demonstrated that allopurinol regressed LV hypertrophy and improved endothelial function in patients with ischemic heart disease There are several limitations in the current study. First, we enrolled cardiac inpatients with a wide variety of cardiac abnormalities; therefore, whether UA represents an independent risk factor for LV hypertrophy in patients with a specific condition―hypertrophic cardiomyopathy, for example―requires further investigation. Second, we excluded patients who were taking UA lowering agents. Whether the association between serum UA and LV hypertrophy might be modified by certain UA lowering drugs should be assessed in future studies.In summary, among male cardiac patients, serum UA was associated with LV hypertrophy independent of intact PTH levels, which showed a significant correlation with serum UA, and other possible confounders, including blood pressure and renal function. Whether strategies leading to UA lowering would improve cardiac performance by decreasing LV mass in cardiology patients awaits further investigation.


Platelet-derived growth factor (PDGF) is a potent stimulator of growth and motility of vascular smooth muscle cells (VSMCs). Abnormalities of PDGF/PDGF receptor (PDGFR) are thought to contribute to vascular diseases and malignancy. We previously showed that a carotenoid, lycopene, can directly bind to PDGF and affect its related functions in VSMCs. In this study we examined the effect of the other naturally occurring carotenoid, lutein, on PDGF signaling and migration in VSMCs.2O2 signaling. Flowcytometry was used to determine PDGF binding to VSMCs. Fluorescence microscopy was performed to examine intracellular ROS production. Modified Boyden chamber system (Transwell apparatus) was used for migration assay.Western blotting was performed to examine PDGF and H2O2-) induced ERK1/2 and p38 MAPK activation. A further analysis indicated lutein could inhibit a higher concentration of H2O2-induced PDGFR signaling, which is known to act through an oxidative inhibition of protein tyrosine phosphatase. Finally, we showed that lutein functionally inhibited PDGF-induced VSMC migration, whereas its stereo-isomer zeaxanthin did not, revealing a special action of lutein on VSMCs.Lutein reduced PDGF signaling, including phosphorylation of PDGFR-β and its downstream protein kinases/enzymes such as phospholipase C-γ, Akt, and mitogen-activated protein kinases (MAPKs). Although lutein possesses a similar structure to lycopene, it was striking that lutein inhibited PDGF signaling through a different way from lycopene in VSMCs. Unlike lycopene, lutein not only interacted with (bound to) PDGF but also interfered with cellular components. This was evidenced that preincubation of PDGF with lutein and treatment of VSMCs with lutein followed by removing of lutein compromised PDGF-induced signaling. Lutein reduced PDGF-induced intracellular reactive oxygen species (ROS) production and attenuated ROS- (HOur study reveals a differential action mechanism of lutein from other reported caroteinoids and suggests a possible beneficial effect of lutein but not zeaxanthin on prevention of vascular diseases. Abnormal vascular smooth muscle cell (VSMC) proliferation and migration play an important role in the development and progression of proliferative cardiovascular diseases (CVDs), including hypertension, restenosis, and atherosclerosis -3.Platelet-derived growth factor (PDGF) is a potent stimulator of growth and motility of connective tissue cells such as fibroblasts and SMCs . PDGF isin vivo [PDGF participates in stimulating SMC proliferation and migration during atherosclerosis . Expressin vivo . PDGF anin vivo . PDGF stin vivo . Overexp2-) and hydrogen peroxide (H2O2). Inhibition of ROS reduces vessel remodeling and restenosis [2O2 at a higher concentration stimulation lead to intracellular ROS production and regulate protein tyrosine phosphatase (PTP), which induces an elevation of tyrosine-phosphorylated proteins [In addition to PDGF, vascular injury also induces oxidative stress and elevated production of reactive oxygen species (ROS) in the vessel wall ,12. Oxidstenosis . Moreovestenosis . It was proteins -18.in-vitro study, lutein and other carotenoids such as lycopene have been shown to reduce adhesion molecules expression in human aortic endothelial cells [Lutein and its stereo-isomer, zeaxanthin, are carotenoids without provitamin A activity and found in a wide variety of fruits and vegetables, including cooked spinach, lettuce, broccoli, peas, lima beans, orange juice, celery, string beans, and squash ,20. It hal cells . This real cells . Howeveral cells , suggestWe previously demonstrated that lycopene inhibits VSMC proliferation and migration through direct interaction with PDGF ,26. The 2O2) was from Merck KGaA Co. . Antibodies (Abs) raised against phospho-ERK1/2 and PDGFR-β were from Santa Cruz Biotechnology . Ab raised against phospho-PDGFR was from Upstate Biotech Inc. . Abs directed against PLCγ1, phospho-PLCγ1 (Tyr783), total p38 MAPK, phospho-p38 MAPK and total Akt were from Cell Signaling Technology, Inc. . Recombinant PDGF-BB and Ab for total ERK1/2 were from R&D systems, Inc. . Lutein and zeaxanthin were purchased from Extrasynthese  and were dissolved in dimethyl sulfoxide (DMSO) and tetrahydrofuran, respectively.The inhibitors for mitogen activated protein kinases (MAPKs) and phosphoinositide-3-kinase (PI-3K), bovine type I collagen and (+/-)-6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid (Trolox) were purchased from Sigma Chemical Co. . Hydrogen peroxide  were plated to the upper chamber in the presence of vehicle or lycopene. After 3 h of incubation, all nonmigrant cells were removed from the upper face of the Transwell membrane with a cotton swab and migrant cells were fixed and stained with 0.5% toluidene blue in 4% paraformaldehyde. Migration was quantified by counting the number of stained cells per × 100 field  under the phase-contrast microscope (Leica DMIL®) and photographed.Migration assay with VSMCs was performed using a modified Boyden chamber model . Briefly, the lower face of polycarbonate filter (Transwell insert) was coated with type I collagen (10 μg/ml) for 30 min in the laminar flow hood. The lower chamber was filled with serum-free, PDGF-BB-containing medium preincubated with vehicle (DMSO) or lutein for 30 min. VSMCs (2.5 × 105) were then stained with biotinylated protein  or biotinylated PDGF-BB at 4°C for 1 h in the presence of lutein or anti-PDGF-BB blocking Ab. After incubated with the fluorescein-conjugated avidin, cells were analyzed immediately by Partec CyFlow ML cytometer  using excitation and emission wavelength at 488 and 525 nm, respectively. Fluorescence signals from 7,500 cells were collected to calculate mean fluorescence intensity of a single cell.Flowcytometric analysis of PDGF binding to VSMCs was performed according to the manufacturer's protocol of the PDGF-BB biotinylated fluorokine kit . Briefly, cells were incubated at 37°C for 1 h to allow regeneration of the receptors. Cells  and incubated at 37°C for 15 min. After washed with PBS, cells were immediately analyzed under the fluorescence microscope  using excitation and emission wavelength at 485 and 525 nm, respectively, and photographed by a digital camera. The fluorescence intensity of the positive staining cells was calculated based on their gray levels (from 0 to 255), as judged by image analysis software of Image-Pro Plus .Intracellular production of ROS was determined by the fluorescence microscopy as previously described  with mint test.Data were expressed as mean ± standard error mean (SEM). Comparison of means of two groups of data was made by using the unpaired, two-tailed Student It has been reported that PDGF-BB binding to PDGFR (PDGF receptor) is associated with activation of PDGFR-tyrosine kinase activity , subsequIn our previous study we have demonstrated that PDGF signaling affected lycopene is not through intracellular inhibition but is through direct interaction with PDGF ,26. Beca2O2-induced signaling in VSMCs. As reported by others, in our experimental system 50 μM of H2O2 could activate MAPKs signaling, including activation of ERK1/2 and p38 MAPK within 30-min stimulation. The activation by H2O2 was inhibited by lutein, whereas total MAPKs were not affected -induced PDGF signaling. It was found that H2O2-induced PDGFR and PLC-γ phosphorylation was inhibited in a concentration-dependent manner.Since intracellular components were affected by lutein during PDGF signaling. Next, we tested whether lutein could affect PDGF-induced intracellular ROS production. As shown in Figure d Figure . Since HIt has been reported that PDGF causes cell proliferation and migration through activation of ERK1/2 and other signaling enzymes ,29. MoreLutein elicited an inhibitory effect on PDGF-BB-induced signal transduction pathway such as PDGFR, PLCγ, MAPKs, and ROS production. Next, we examined whether lutein functionally affected PDGF-BB-induced VSMC migration. As shown in Figure 2O2)-induced signaling and a higher concentration of H2O2-induced signaling -epigallocatechin-3-gallate (a tea polyphenol) ,32, luteg Figure . In strig Figure . Therefo2O2 stimulation lead to intracellular ROS production and regulate protein tyrosine phosphatase (PTP), which induces an elevation of tyrosine-phosphorylated proteins [2O2 (50 μM)-induced ERK1/2 and p38 MAPK activation in VSMCs   Figure , which iIt is an interesting issue that the carotenoids with a similar structure act in a differential way on VSMCs. Our previous studies have shown that lycopene affects PDGF signaling through interaction with PDGF but not cellular components ,26. HoweIn this study we provided evidence that lutein interacts with PDGF and affects cellular components, leading to interference with PDGF intracellular signaling, and functionally inhibits VSMC migration. We also demonstrated that zeaxanthin, the stereo-isomer of lutein, shows a distinct effect on PDGF signaling and VSMC migration. Our findings, together with our previous study, not only suggest the possible beneficial effect of lutein in preventing cardiovascular diseases but also an intriguing phenomenon of these carotenoids in inhibiting of PDGF signaling, highlighting their differences in mechanism of action in affecting VSMC behaviors.CVD: Cardiovascular disease; ERK: extracellular matrix-regulated kinase; PDGF: platelet-derived growth factor; PDGFR: PDGF receptor; PLC-γ: phospholipase C-γ; MAPK: mitogen-activated protein kinase; VSMC(s): vascular smooth muscle cell(s).The authors declare that they have no competing interests.HML and WBW conceived of the study and designed research. HML, YJT, WYD and CJT performed research and analyzed data. WBW wrote the paper. All authors read and approved the final manuscript.


Primary cilia have recently been highlighted as key regulators in development and disease. This review focuses on current work demonstrating the broad role of cilia-related proteins in developmental signaling systems. Of particular consideration is the importance of the basal body region, located at the base of the cilium, in its role as a focal point for many signaling pathways and as a microtubule organizing center. As the cilium is effectively a microtubular extension of the cytoskeleton, investigating connections between the cilium and the cytoskeleton provides greater insight into signaling and cell function. Of the many signaling pathways associated with primary cilia, the most extensively studied in association with the cytoskeleton and cytoskeletal rearrangements are both canonical and non-canonical Wnt pathways. One of the key concepts currently emerging is a possible additional role for the traditionally 'cilia-related' proteins in other aspects of cellular processes. In many cases, disruption of such processes manifests at the level of the cilium. While the involvement of cilia and cilia-related proteins in signaling pathways is currently being unraveled, there is a growing body of evidence to support the notion that ciliary proteins are required not only for regulation of Wnt signaling, but also as downstream effectors of Wnt signaling. This review summarizes recent advances in our understanding of the involvement of cilia and basal body proteins in Wnt signaling pathways. The primary cilium had long been ignored by biologists and was considered to be a vestigial remnant of evolution, similar to the human appendix. However, a growing number of human disorders are being attributed to dysfunctions in cilia or cilia-related proteins , leadingMore recently, the primary ciliary membrane has been shown to be rich in various channels and receptors ,5, suggeThis review will highlight our current understanding of the cilium and basal body as one of the microtubule organizing centers of the cell and how they function as a critical signaling center. The cilium has been implicated in the regulation of a growing number of signaling pathways, and while much recent research has focused on sonic hedgehog (Shh)-signaling, this review will more closely examine the role of the cilium and basal body in regard to Wnt signaling, an as yet less explored avenue of research. To begin, we will take a closer look at the cilium and related proteins, as well as, the possible roles of those proteins beyond the cilium. Then, the role of cilia and basal bodies in both canonical and non-canonical Wnt signaling will be examined; focusing more specifically on their association with cytoskeletal regulation.Cilia are microtubule-based organelles, extending from the plasma membrane and coupled to the cytoskeleton. They are anchored to the cell and emanate from the basal body, a barrel structure of nine triplet microtubules that elongate to form the microtubule doublets of the ciliary axoneme. The basal body is derived from the mother centriole, and is also considered a microtubule-organizing centre (MTOC). Centrioles are more than just the primary MTOC of a cell; they also serve as important regions for protein interactions, and as docking stations for proteins on the way to the cilium ,9. The rIn eukaryotic cilia the transition zone is composed of several distinct structures including transitional fibers, Y-linkers, and the ciliary necklace (on the cilia membrane), all flanked by the terminal and basal plates. While their appearances vary among species, the basic components are conserved . Though Sensory cilia are additionally specialized by cell type to transduce the specific function or modality of that particular cell. For example, olfactory cilia detect odorants via specific receptors on the ciliary membrane, the connecting cilia in mammalian photoreceptors continuously transport various proteins to support the outer segments, and sensory cilia lining the kidney tubules sense fluid flow through the nephron.CEP290 can cause nearly all ciliopathies [BBS4 or NPHP5 are restricted to two [Initial identification of disease causing genes in ciliopathies such as Bardet-Biedl syndrome (BBS) or NPHP, suggested that each ciliopathy had a distinct set of disease-causing genes. However, in recent years it has become apparent that there is considerable overlap between ciliopathy genes, suggesting that phenotypic variability among ciliopathies is caused by a combination of the nature of the mutation and the contribution of other allelic variations. Mutations in genes such as opathies ; others d to two . In gened to two ,18,21.The theory that 9+2 cilia solely mediate motile functions, while the 9+0 cilia have a strictly sensory function is now considered an oversimplification. Non-motile olfactory sensory cilia have a 9+2 structure , while tAs the structural basis of all cilia is a ring of nine microtubules, ciliopathies could alternatively be viewed as diseases of microtubule function, with the cilium representing a specialized extension of the cytoskeleton. Whereas microtubule defects inside the cell might be mitigated due to compensatory roles of other proteins, the highly specialized ciliary transport machinery may be particularly sensitive to perturbations in microtubule function/transport due to its remote location and lack of compensatory subunits. Many of the ciliopathy genes code for proteins that localize to the basal body and/or transition zone and, as such, may exert a regulatory role on ciliary function. As microtubules are highly abundant in this region, their association and interaction with ciliary proteins are essential for ciliary function. In addition, we must consider the possibility that proteins important for cilia-related functions may also be required for non-ciliary microtubule and cytoskeletal associated processes -29.If the cilium is regarded as a specialized extension of the cytoskeleton, it is not surprising that many ciliary proteins are also beginning to emerge as key players in other cellular processes that involve the cytoskeleton. These include cell cycle progression and vesicular transport, both of which are heavily dependent on the configuration of the microtubule architecture within a cell.The basal body, which anchors the cilium, is one of the microtubule-organizing centers of the cell, and recent work has identified a crucial link between ciliogenesis and cell cycle progression (reviewed in Kim and Tsiokas ). In celAs the basal body derives from the mother centriole, the timing of ciliogenesis is closely related to mitosis, mitotic spindle deconstruction, and centriole duplication. Cilia customarily nucleate and grow during the G1-G0 phase of the cell cycle and then retract during mitosis . The cenDrosophila neuroblasts and male germline cells [vs. younger centriole of dividing sister cells adds a further layer of complexity to signaling regulation [Asymmetric centrosomal inheritance has been implicated in subsequent cell fate decisions in ne cells -43. In mne cells . Cells igulation . AsymmetThese findings, namely that asymmetric distribution of proteins around the centriole is also important during both cell cycle progression and ciliogenesis, support the concept that the basal body region plays a critical regulatory role. Other contributors to such functions include the ciliary-related proteins CEP290, CP110, and Cep97, which are highly involved in cilia assembly and regulation and are also asymmetrically localized to the basal body region ,44,45. AIn the emerging association of cilia and cell cycle progression, two dynein binding proteins, Ned1 and Tctex-1, have been shown to regulate cell cycle progression via effects on cilia length ,49. BothAs IFT proteins have been shown to be required for intracellular trafficking to the cilium, not just along the cilium, it is plausible that these components might also be required for additional microtubule related transport. Intriguingly, the intraflagellar transport protein IFT88, required for ciliary anterograde transport, has also been shown to be required for spindle orientation during mitosis ,50, thusin vitro evidence is mounting that many ciliary proteins have additional, alternate roles away from the cilium, it is important to note that in vivo genetic evidence is as yet lacking. Many mouse and zebrafish mutants which lack cilia appear to survive well into development [While elopment ,52. NeveIn recent years, many different signaling pathways, such as canonical and non-canonical Wnt, Shh53--53-57, FWhile both canonical and non-canonical Wnt signaling are activated by Wnt molecules binding to distinct membrane-bound receptors, downstream steps diverge. Canonical Wnt signaling results in stabilization of cytoplasmic β-catenin, which subsequently enters the nucleus and initiates transcription of Wnt target genes. In contrast, downstream events of PCP signaling result in changes in cell morphology  rather than transcription. This process ultimately results in the directed orientation of cells relative to a planar axis within the epithelium. While the precise role of cilia in Wnt signaling is currently being dissected , a key ssitus inversus [Kif3a, Ift88, Ofd1) resulted in hyperactive canonical Wnt responses [One of the first indications that cilia might be involved in Wnt signaling was the identification of two key PCP components, Inversin (Nephrocystin 2) and Dishevelled (Dvl), at the basal body Table  47,65,6,665,66. inversus . This waesponses -70. Mutaesponses -75. ThesIft88, Ift72, and Kif3a mutant mouse embryos, all of which lack primary cilia, show normal expression of the canonical Wnt target gene, Axin2, and normal activation of a transgenic canonical Wnt reporter [It is also important to note that there have been some studies that suggest there is no connection between cilia and Wnt signaling. Maternal-zygotic ift88 zebrafish mutants lack all cilia, yet display normal canonical and non-canonical Wnt signaling . At midgreporter . Mouse ereporter . HoweverDrosophila and includes polarized localization of core PCP proteins, which ultimately leads to the establishment of polarized cell morphology [The PCP signaling pathway was initially described in rphology ,77. In vrphology , defectsrphology  and hairrphology . One of rphology ,82. In rrphology ,83-86.As discussed, initial observations of PCP defects in ciliary mutants  ,82 suggePkd1, Pkd2, and Pkhd1 are conflicting and suggest that loss of oriented cell division alone does not initiate cyst formation. While oriented cell division is disrupted in Pkhd1 mutants, this alone appears not to be sufficient to produce kidney cysts. In contrast mutations in Pkd1 and Pkd2 do result in cyst formation despite normal oriented cell division, suggesting that loss of PCP-related oriented cell division is not sufficient to generate kidney cysts [In the mammalian kidney, cilia/basal body dysfunction has also been suggested to affect the orientation of mitotic spindles, associated with defective tubule division and resultant polycystic kidney disease . Defectiey cysts . Therefo-/- Fat4mice display severe PCP phenotypes thought to be caused by abnormal orientation of mitotic spindles [Ciliary localization of several core PCP proteins adds a further layer of complexity to dissecting this relationship. The protocadherin Fat4 was shown to localize to the cilium, and spindles . Two othspindles , Vangl2 spindles . Vangl2 spindles  and in cspindles . In addispindles . Fluid fspindles -98.Xenopus and Drosophila PCP effectors Inturned and Fuzzy are responsible for ciliogenesis and have been shown to affect apical actin assembly and microtubule polarity but not nucleation. Defects may occur in cytoskeletal assembly that impair movement of basal bodies and centrioles [Xenopus embryos. A recent study found an enrichment of non-synonymous coding changes in human Fritz in two ciliopathies: Bardet-Biedl and Meckel-Gruber syndrome [It is possible that correct basal body and centrosome positioning could be required for subsequent regulation of cytoskeletal architecture. The basal body, as a MTOC, could regulate the nucleation and outgrowth of microtubules; indeed, several of the BBS proteins have been shown to participate in microtubule nucleation and growth ,99. Evidntrioles . The PCPsyndrome . Of notesyndrome .Smurf mutant cochlea, polarity proteins (Prickle 1) are not asymmetrically degraded at the region of the basal body as in wild-type animals. This further implicates the basal body as a regional signaling center assisting the activation and/or degradation of signaling proteins. A recent proteomic screen found several PCP-associated proteins in the human centrosome such as PRICKLE3, SCRIB, CCDC66, and Albatross [As will be discussed in subsequent sections, regulation of various signaling molecules (including Rho A and Dvl) could occur at the basal body. Examples of this include the Smurf ubiquitin ligases, which target proteins for degradation by proteasomes. These proteins regulate PCP signaling via localized Prickle degradation by the Par6-Dvl2-Smurf complex . In Smurlbatross . This sclbatross ,104.Similar to the previous quandary regarding PCP and centrosome/basal body positioning, a key question that remains to be elucidated is which comes first: is intact PCP signaling required for ciliogenesis, or is an intact cilium necessary for correct PCP signaling? As downstream effects of PCP signaling are associated with actin and microtubules, one could envisage a possible feedback loop where intact cytoskeletal architecture and function is in turn necessary for cilia formation or function. Another key question is whether the cilium itself is responsible for the defects observed in some PCP mutants. Many of the cilia-related proteins are now being identified in other cellular locations and could possibly be required for additional intracellular functions. As suggested before, the cilium is heavily dependent on ciliary proteins, and defects in their function may be more readily observable at this organelle.Jouberin, mutations of which cause the ciliopathy Joubert syndrome, has been shown to facilitate β-catenin translocation to the nucleus [At first glance, the β-catenin-dependent canonical Wnt signaling pathway appears relatively straightforward. Wnt ligands bind and activate the Frizzled class of receptors, thereby stabilizing β-catenin, which then enters the nucleus and subsequently activates Wnt target genes . This pa nucleus . As willAs we further discuss the various components required for Wnt signaling and their association with cilia and basal bodies, one factor to consider is which is more dependent on the other: is intact cytoskeletal architecture  required for Wnt signaling, or is Wnt signaling required for the establishment of a fully functional cytoskeleton encompassing a cilium and basal body?vs. daughter centrioles and may play a role in primary cilia formation and function [Chlamydomonas reinhardtii have identified EB1 localizing to subdistal appendages of the mother centriole/basal body [The tumor suppressor protein APC is one of the key components in the canonical Wnt signaling pathway. Most of the scientific literature is focused around its ability to bind and induce degradation of β-catenin. However, there are additional roles for this protein involving the organization of cytoskeletal networks . APC hasfunction . This mofunction . The EB function . Interesfunction . Studiessal body ,117, whisal body ,118.As the centrosome and basal body region at the base of the cilium is beginning to emerge as a cellular signaling centre, it is not difficult to imagine that APC, in its role as part of the β-catenin destruction complex, might be anchored to this region. In support of this theory, other related components are also being identified in this area such as members of the kinesin class of motor proteins.The kinesins, a class of motor protein, exert a regulatory effect on Wnt signaling. Many proteins that bind to microtubules participate in the regulation of microtubule dynamics. These include the kinesins, which are powered by the hydrolysis of ATP. Although kinesins share similar motor domains, they exhibit a variety of functions, including regulation of microtubule dynamics. Kinesins are known to be crucial for ciliogenesis and intraflagellar transport ,119. Rec99, MCAK dysfunction could hinder correct localization of APC, thereby disrupting Wnt signaling.Wnt signaling at the centrosome is one of the ways in which kinesins exert regulatory functions. Not only are kinesins required for localization of APC to the centrioles and centrosome, but another kinesin, MCAK , also associates predominantly with these structures. As a member of the kinesin-13 family (other members of which include KIF2A and KIF2B), MCAK has been shown to be critical for microtubule dynamics and organization . This prMCAK has also been shown to associate with the microtubule-associated protein EB1 via its C-terminus at growing ends of microtubules ,125. WheIn summary, kinesins are not only required for trafficking of structural or signaling components to and along the cilium, but are also required for stabilization and regulation of the cytoskeletal infrastructure along which these molecules travel. In these capacities the kinesins are able to exert significant control over various cellular mechanisms.Identification of ciliary proteins that affect both canonical and non-canonical Wnt signaling are not surprising, considering that the cilium can potentially be seen as a branch point bisecting the two pathways. These include a family of proteins, the GTPases. The GTPases are structurally related to kinesins, however, they hydrolyze GTP instead of ATP. GTPases are involved with cellular trafficking, and several members of the ARF-like family of small GTPases have been implicated in ciliary function ,127. As Cdc42, a Rho family GTPase, is a key player in Wnt-dependent cytoskeletal rearrangements and a key regulator of cell polarity. Cdc42 acts as a molecular switch, modulating a wide range of signaling pathways. In cellular trafficking, Cdc42 functions at several steps of endocytosis, recycling, and biosynthesis, influencing polarized trafficking in epithelial cells . In its Importantly, the Cdc42/Par6/aPKC complex has been localized to the primary cilium and has been shown to be necessary for ciliogenesis in mammalian cells ,135. WhaC. elegans, the small GTPase RAB5 has been shown to localize to the base of the cilium and regulate polycystin 1 and 2 removal from the cilium [Cdc42 is not the only GTPase associated with the cilium. Other GTPases that also localize to the cilium could function in a similar way to, or in conjunction with, Cdc42. One such GTPase is ARL6/BBS3, one of the BBS proteins not associated with the core group of BBS proteins comprising the IFT-participating 'BBSome', a stable protein complex which functions in primary cilia biogenesis . This sue cilium .Other proteins that appear to intercept the two signaling pathways include pVHL. The pVHL protein has been proposed to control ciliogenesis by orienting microtubule growth. This protein is a cargo of Kif3 and has been shown to interact with the polarizing PAR3-PAR6-aPKC complex, suggesting that these proteins may operate in the same pathway to regulate cortical growth of microtubules and formation of cilia . PreciseIdentification of molecules that are already known to function as molecular switches between signaling pathways, suggests that there are possibly more awaiting discovery. These may also have the capacity to control or 'fine tune' Wnt signaling decisions. The basal body, an emerging signaling 'hotspot' perfectly located at the base of the 'sensory-loaded' cilium might be an ideal cellular location for the switch.We are only beginning to understand the role of the cilium and its biological influence. The cilium is more than just a microtubular appendage anchored to the cell via the basal body. The multiple parts of the cilium from the basal body to the distal tip Figure  are justThe base of the cilium is the most crucial part of this 'communication device', as the incoming signals need to be ordered and regulated. It is perhaps not a coincidence that the basal body doubles as the MTOC of the cell. The basal body is most likely regulating other internal signaling and trafficking events as well. Regulation of this region is a hugely critical and as yet underexplored area. However, many kinases, phosphatases, kinesins, and proteosomal proteins that localize to this region are being identified. Of particular consideration in regard to cilia, two of the kinases known to affect Wnt signaling, CKID and PPPIC, have been identified in the ciliary proteome .C. elegans [The base of the cilium is not just a signaling center, but also a sorting center. An emerging concept is that of scaffold proteins acting as hubs for controlling the flow of cellular information  within a cell . Several elegans . This isThe concept of the cilium as a potential regulator (molecular switch) between canonical and non-canonical Wnt signaling has been suggested  and coulIt would certainly be an oversimplification to assume that a single type of cilium would suffice for the needs of every cell. The key to our understanding of cilia function will be to distinguish the characteristics of cilia in differing cells and tissues. The recent explosion of ciliary research is likely to continue for the foreseeable future as this long forgotten organelle reveals its inner secrets.APC: adenomatosis polyposis; CK1: Casein kinase 1; BBS: Bardet-Biedl syndrome; GSK3β: glycogen synthase 3b; IFT: intraflagellar transport; NPHP: nephronophthisis; PCM: pericentriolar material; PCP: planar cell polarity; Shh: Sonic hedgehog.The authors declare that they have no competing interests.HMS prepared the manuscript, MK edited the manuscript. Both authors read and approved the final manuscript.


Metabolic syndrome (MetS) and hyperuricemia are important risk factors for cardiovascular disease. However, findings regarding the relationship between serum uric acid (UA) level and components of MetS have been inconsistent. This study was performed to explore the potential value of UA level as a marker of MetS among male and female Chinese of different ages.A total of 5896 subjects  were recruited from the Department of Health Management at the Chang Gung Medical Center. Hyperuricemia was defined as a serum UA value >7.0 mg/dL for males or >6.0 mg/dL for females. MetS was defined according to the criteria of the Adult Treatment Panel III, as modified for Chinese subjects. Serum UA was used to differentiate MetS and to calculate epidemiological indices by means of discriminate analysis and logistic regression.P < 0.01). A positive correlation between serum UA and MetS was observed in both sexes. Serum UA and the occurrence of MetS rose with increasing age in females; in males, however, UA values did not vary with age.The sensitivity and specificity of serum UA concentration as a marker of MetS ranged from 55.2% to 61.4% and 61.9% to 68.4%, respectively. Subjects with high UA had a higher risk of MetS, with odds ratios ranging from 1.23 to 1.82 (Serum UA is more closely associated with MetS in females than in males. High UA among middle-aged women may predict the development of MetS. Furthermore, it is unclear whether hyperuricemia increases with age or is sex-specific in Chinese. The present study evaluated Taiwanese adolescents and adults for hyperuricemia and risk factors for MetS to ascertain whether direct correlations between these parameters were a function of age and/or sex.Hyperuricemia and the risk for MetS may vary with age and sex. For example, gouty arthritis and cardiovascular complications are rarely observed in premenopausal females; however, the incidences of hyperuricemia and MetS increase dramatically after menopause.Subjects for this investigation were recruited from the Department of Health Examination at the Chang Gung Memorial Hospital in Kweishan, Taoyuan, Taiwan during the period from 2003 through 2005. All subjects in this study were Chinese residents of Taiwan. The study group comprised 5896 subjects, including 2960 females and 2936 males. The average age of the participants was 53.9 ± 12.0 years ; the average age was 53.5 ± 11.5 years  for females and 54.4 ± 12.5 years  for males. Because some of subjects recruited from the Department of Health Examination were undergoing examinations for health problems, the authors acknowledge the presence of bias in this study.Different methods were used to evaluate the association between serum UA and MetS and to differentiate MetS as a function of UA concentration. UA values were used to divide the subjects into subgroups according to sex. The UA values (in mg/dL) for the 10 subgroups of females were 3.8 or lower, 3.9 to 4.3, 4.4 to 4.6, 4.7 to 5.0, 5.1 to 5.3, 5.4 to 5.6, 5.7 to 6.1, 6.2 to 6.6, 6.7 to 7.4, and 7.5 or higher. The corresponding UA values for the 10 subgroups of males were 4.9 or lower, 5.0 to 5.5, 5.6 to 5.9, 6.0 to 6.3, 6.4 to 6.7, 6.8 to 7.1, 7.2 to 7.6, 7.7 to 8.1, 8.2 to 9.1, and 9.2 or higher.17,18 The average ages of the young, middle-aged, and elderly groups were 37.0, 53.3, and 70.8 years, respectively, for females and 37.6, 54.0, and 71.4 years, respectively, for males.Age is considered the most important factor affecting UA concentration and the development of MetS. Therefore, subjects between 26 and 75 years of age were allocated to subgroups based on 5-year age-bands. Subjects were also divided into 3 standard age groups: young (44 years or younger), middle-aged (45–64 years), and elderly (65 years or older).4,19 MetS was defined according to the criteria of the modified Adult Treatment Panel (ATP) III issued in 2004 by the Bureau of Health Promotion, Department of Health, ROC (Taiwan). Subjects satisfying 3 or more of the following criteria were defined as having MetS: central obesity with a waist circumference >90 cm for males or >80 cm for females, or BMI ≥27 kg/m2; triglyceride ≥150 mg/dL (1.695 mmol/L); high-density lipoprotein (HDL) cholesterol <40 mg/dL (1.036 mmol/L) for males or <50 mg/dL (1.295 mmol/l) for females; blood pressure ≥130/85 mm Hg; and fasting glucose ≥100 mg/dL (≥5.6 mmol/L).19 The study was approved by the Institutional Review Board of Chang Gung Memorial Hospital. Subjects with an obvious disease or disorder, or a weight change exceeding 10%, during the 6 months before initiation of the study were excluded.Venous blood samples were obtained after an overnight fast and subjected to centrifugation at 3000 rpm for 30 minutes at 4°C. UA present in blood samples was measured by a colorimetric enzymatic method. Hyperuricemia was defined as a UA value above 6.0 mg/dL for males or above 7.0 mg/dL for females.17 Logistic regression was used to analyze the odds ratio (OR) of UA by using the presence of MetS as the dependent variable. P values <0.05 were considered statistically significant.All data analyses were performed using SPSS software. Separate statistical analyses were performed for males and females and for each age group. Normal variables were evaluated by using the chi-square test for the presence or absence of MetS and hyperuricemia. Discriminate analysis was used to test the correlation between MetS and UA. In addition, serum UA concentration was utilized to distinguish MetS and to calculate sensitivity, specificity, pre-test probability , positive likelihood ratio (LR+), and negative likelihood ratio (LR−). The following formulas were used to calculate the parameters relevant to this analysis : sensitivity = a/(a + c); specificity = d/(b + d); LR+ = sensitivity/(1 − specificity); LR− = (1 − sensitivity)/specificity; pre-test probability  = (a + c)/(a + b + c + d); and post-test probability = post-test odds/(post-test odds + 1), with post-test odds = [a/(a + b)] * [prevalence/].The percentages of female and male subjects with MetS in each of the 10 subgroups of UA concentration are shown in Figure Figure P < 0.05), except for the category of male subjects with or without hyperuricemia. These findings indicate that among these 3 age groups, the distributions of both MetS and hyperuricemia differ significantly. Of the 631 male subjects in the young group, the 1652 in the middle-aged group, and the 653 in the elderly group, 146 (23.1%), 524 (31.7%), and 194 (29.7%), respectively, satisfied the criteria for MetS. In females, age was a major factor influencing MetS: 57 (10.8%), 521 (27.3%), and 262 (50%) of females had MetS in the young, middle-aged, and elderly groups, respectively. Age and sex differences were also observed when the other parameter, hyperuricemia, was examined. A total of 119 (22.5%) females and 293 (46.4%) males in the young group, 622 (32.6%) females and 720 (43.6%) males in the middle-aged group, and 226 (43.1%) females and 282 (43.2%) males in the elderly group had hyperuricemia. Although the incidence of hyperuricemia, like that of MetS, differed with age and sex, the incidence was higher for male subjects than for female subjects in the young and middle-aged groups.We further analyzed the effects of age on MetS and serum UA concentration by reducing the age groups to 3—young, middle-aged, and elderly subjects—for both females and males. These groups were then divided into subgroups based on the absence or presence of hyperuricemia or MetS Table . SignifiP < 0.01 for all) and was highest for young males. Among females, this coefficient ranged from 0.25 to 0.32  and was highest for middle-aged women.Table The sensitivity of serum UA concentration for the diagnosis of MetS ranged from 58.0% to 61.4% in females and from 55.2% to 58.9% in males. The highest specificities were observed in young females and young males—68.4% and 66.0%, respectively. The lowest specificities were observed in elderly subjects—65.3% for women and 61.9% for men. In both sexes, the likelihood ratios for a positive test result were higher in the young and middle-aged groups than in the elderly group, especially for young females (1.94). For males, the likelihood ratios for a negative test result were similar in all 3 age groups and ranged from 0.62 to 0.72; for females, these ratios ranged from 0.56 to 0.64. Regarding pre-test probabilities, this index rose sharply with increasing age for females with MetS: 10.8%, 27.3%, and 50.0% for young, middle-aged, and elderly subjects, respectively. In male subjects, the highest prevalence was 31.7% in the middle-aged group and the lowest was 23.1% in the young group. Post-test probabilities, however, were lower than prevalences in all populations. The post-test probability for high serum UA concentration in the identification of MetS was almost 38.5% in elderly females, and the deviation between prevalence and post-test probability indices ranged from 7.7% to 15.9%.3,18 However, UA concentration was more closely associated with MetS in females than in males,16 and the highest risk of MetS was observed in middle-aged women with high UA.This study was performed to evaluate serum UA concentration as a marker of MetS among Chinese males and females of different ages. We found that high UA was associated with an increased risk of MetS.18 the prevalence of hyperuricemia was found to be significantly higher among subjects with MetS, as compared with those without MetS. High UA has also been found to correlate with several risk factors for MetS19; in adult populations, elevated serum UA was independently correlated with these risk factors.10 Furthermore, UA was found to rise as the number of components of MetS in a person increased.20 Sex-specific distribution of the occurrence of MetS among subjects with different serum UA concentrations has also been observed. In a prospective analysis of the predictive value of hyperuricemia in residents of the United States,4 males with a serum UA value ≥6.5 mg/dL were found to have a 1.60-fold increased risk of MetS as compared to males with a concentration <5.5 mg/dL; in females with a serum UA concentration ≥4.6 mg/dL, the risk was at least 2-fold higher. In recent studies describing an association between hyperuricemia and the risk of MetS, the odds ratio was found to be similar for males and females when hyperuricemia was defined as a UA level of 6 mg/dL for females and 7 mg/dL for males.21 Lastly, a close association of hyperuricemia with MetS has been observed among different ethnic groups.2,3,5These findings are in good agreement with those of previous studies. In this study, as in a recent study of elderly Taiwanese,Because the incidence of MetS was relatively low in females and males with low UA, a UA value below the average represents a low risk for MetS. A UA value higher than average but below the cutoff value for hyperuricemia should be considered medium risk for MetS. In the present study, the incidence of MetS was actually much higher among subjects in the medium-risk category than among those in the low-risk category. A UA value in the hyperuricemic range should be regarded as high-risk because the incidence of MetS was markedly higher at these concentrations, especially in females.19,22,23 In the present study, the most important observation regarding sex was that the incidence of MetS in females with lower-than-average UA was significantly lower than that in males with the same UA values. Overall, the findings of the present study agree with those of a recent study5 in which serum UA was found to be associated with metabolic syndrome and some of its components in Japanese. In that study, however, rates of hyperuricemia were similar in elderly men and women, but women with hyperuricemia had higher rates of MetS than did men with hyperuricemia.Sex is clearly an important factor in the inter-relationships among hyperuricemia, MetS, and age. A limitation of the present study was the absence of information on the menopausal status of women. However, in other investigations, the rates of MetS and hyperuricemia were consistently higher in women of postmenopausal age than in premenopausal women.19,22,23Analyses examining both sex and age revealed that the incidence of MetS differed between men and women aged 51 to 55 years and 56 to 60 years, and that the age of 55 years was found to be critical for women and men when these subjects were grouped by 5-year increments. Other findings indicate that sex significantly influences serum UA and the occurrence of MetS during aging.14 This discrepancy may be due to differences in lifestyle, eating habits, and health status among the subjects of these studies.24 In the present study, the percentages of subjects with MetS and with hyperuricemia rose significantly in middle-aged and elderly women, but not in men, most likely because MetS and its components develop more frequently after the onset of menopause.22 Indeed, as noted above, postmenopausal women have consistently higher rates of MetS and hyperuricemia than do premenopausal women.13,18Additional analysis disclosed that the distribution of male subjects with or without hyperuricemia did not differ among the 3 major age groups. This result is at odds with a previous study, which showed that serum UA rose with increasing age.This study is the first to use discriminate analysis to examine the association between MetS and serum UA concentration in males and females in different age groups. The post-test odds of MetS were determined by the likelihood ratio, the prevalence of MetS, the characteristics of the subject pool, and information regarding individual subjects. Identification of MetS by serum UA was better in females than in males. Moreover, the coefficient of correlation was higher in females than in males.In conclusion, serum UA values differ significantly with age and sex in Chinese. Serum UA concentration correlates positively with MetS in these subjects, and is more closely associated with MetS in females than in males. These findings provide a very useful predictor of MetS in middle-aged women. Further prospective studies that investigate the mechanism(s) by which sex influences the correlation between serum UA and MetS are recommended.


Human T-cell leukemia virus type 1 (HTLV-1) is an oncogenic retrovirus etiologically causal of adult T-cell leukemia (ATL). ATL is a highly virulent cancer that is resistant to chemotherapeutic treatments. To understand this disease better, it is important to comprehend how HTLV-1 promotes cellular growth and survival. The human T-cell leukemia virus type 1 (HTLV-1) basic leucine zipper factor (HBZ) gene is encoded by the minus strand of the HTLV-1 provirus and transcribed from the 3' long terminal repeat (LTR). HBZ expression promotes proliferation and survival of HTLV-1 infected cells in vivo. Although previous reports have been aimed at understanding the potential role of HBZ in HTLV-1 pathogenesis, little is known as to how this viral gene is regulated in ATL cells. Here using our K30-3’asLuc reporter construct, we show that HBZ protein upregulates antisense transcription. Generation of stable clones in 293T cells confirm that HBZ stimulates antisense transcription from the 3’ LTR through its interaction with JunD, an AP-1 protein, and its action on the Sp1 binding site located in the 3’LTR. Our results suggest that in vivo inhibition of JunD could be a possible new therapeutic strategy in ATL treatment.


Literature review has shown that some years back the fertility desires of people living with HIV was low but in the recent years, it was reported as increasing. However, little is known about the strength of association of fertility desire of HIV positive people with antiretroviral therapy (ART) experience, age, sex, education level, and number of children.2, Chi-square (Q), I2 and P-value. Sensitivity analysis and funnel plot were done to assess the stability of pooled values to outliers and publication bias, respectively.In these meta-analyses, twenty studies from different parts of the world were included. The odds ratios of fertility desires were determined using the random-effects model. Heterogeneity among the studies was assessed by computing values for TauThe pooled analysis demonstrated that fertility desires of study participants had no association with ART. Similarly, the overall odds ratio did not show statistically significant association of fertility desires with sex and educational attainment of study participants although forest plots of some studies fall on increased and some others on decreased sides of fertility desires. The two variables that demonstrated a strong association with fertility desires were age less than 30 years and being childless. The lowest heterogeneity was found in a meta-analysis comparing ART experienced and ART naïve HIV positive people. In all meta-analyses, the sensitivity analyses showed the stability of the pooled odds ratios; and the funnel plots did not show publication or disclosure bias.Although the fertility desires among childless and younger age group was very strong, we realized that quite a significant segment of HIV-infected people have desire for fertility. Therefore, including fertility issue as integral part of HIV patient care may help several of them in their reproductive decision making (letting them know the risks and methods of prevention while anticipating pregnancy). Before the era of HIV, wishing to have a biological child and preserving the continuity of their family to the future generation is perhaps a common thought of all and this is also expected to be same in these days in the majority of individuals who are sero-negative for HIV. However, several studies have shown that the fertility desire of people living with HIV was found to be significantly reduced -4. A natOn the other hand, the fertility desire of HIV-infected individuals were reported as increasing because of increasing hope of living longer with ART, resumption of a healthy life, and the overall physical and mental well-being. A multivariate analysis from Uganda showed that the fertility desire of HIV-infected individuals had significantly increased with ART . In CanaSeveral non-comparative studies have also shown the independent association of other factors with fertility desire in HIV-infected population. A partner’s desire for a child was one of the strongest predictors for women’s fertility desire . In anotIn general, a systematic review including twenty nine studies has also outlined the presence of many more factors that are related to fertility desires . HoweverComputer based search for articles related to the fertility desire of HIV positive people was conducted in Medline, HINARI, google scholar and Cochrane library. Via HINARI, we have also searched the websites of major publishers like: Elsevier Science-Science Direct, Nature Publishing Group, Oxford University Press, PsycARTICLES, Science and Wiley-Blackwell. The search was supplemented by searching the reference lists of each retrieved article. Article search was performed by both authors (YB and AB) with an alternate combination (and/or) of these search terms: “fertility desire”, “fertility intention”, “desire to have children”, “parenthood”, “motherhood”, “maternity paternity”, “people living with HIV”, “HIV” and “ART” or “HAART”.These meta-analyses involved studies with the following inclusion criteria: 1) studies which reported the fertility desire of HIV positive individuals in relation to one or more of the selected variables ; 2) studies which were published in English; and 3) studies done from year 2000 up to June 2012. Studies were selected if they met any of these variables.From the selected studies, the following information was abstracted: author, year of publication, country where the research conducted, sample size, number of HIV positive people having fertility desires . Some variables were dichotomized as no child vs one and more, age less than 30 vs 30 years and above, primary or no education vs secondary and above. For these analyses, data on both fertility intention and desire were similarly entertained.2, Chi-square (Q), I2 and P-value. When the value of I2 was greater than or equal to 50%, the variation across the studies was considered as statistically significant. Sensitivity analyses were done to assess the stability of pooled values to outliers. Publication/disclosure bias was evaluated by funnel plot. All the analyses were done using Meta Analyst (Beta 3.13) software [Five meta-analyses were done taking ART experience, age, sex, education level, and number of children as independent variables. The odds ratios of fertility desires were determined using the DerSimonian-Laird method (Random effects model). Heterogeneity among the studies was assessed by computing values for Tausoftware . P-valueAs shown in Figure 2 = 35.1%).As presented in Figure 2 = 91.7%).However, having no child was a strong predictor of fertility desire ,33,34,372 = 48.9%).In Figure 2 = 93.8%).In Figure Except one study , the metThese meta-analyses demonstrated that the fertility desire was highest among young and childless people living with HIV but was not associated with ART or educational level. ART is known to improve the overall well-being of HIV-infected persons  and manyHowever, as these meta-analyses showed, neither their better education attainment nor being on ART was found to have an influence on their fertility desire. Rather, fertility desire was strongly associated with being young or childless, which both were likely to be interlinked one with the other and this kind of reflection is probably the reflection of the majority of human being in the fertile age regardless of HIV infection. In another study not included in this meta-analysis, being young or having few or no child was also an independent predictor for high fertility desire . To the In general, it is a known fact that identifying the factors behind the fertility desires has paramount importance for both policy makers and health care providers. Specifically, although these meta-analyses did not demonstrate the associations of fertility desire with ART, as majority of HIV positive people are in the fertile age and living longer with ART ,41, the This is despite the fact that fertility desires of people living with HIV are serious concerns for the patients themselves, for authors in the field and health care providers ,15,40,43Therefore, understanding the epidemiology of fertility desires of people living with HIV helps clinicians aware of their clients demand ahead of time and gets prepared to plan and implement the preventive modalities for both horizontal and vertical HIV transmission. This is to mean that letting the couples know the potential health risks to themselves and their babies help them make an informed decision, which is also the recommendation of other authors ,44. WhenThese meta-analyses are not without limitations. Firstly, the duration of ART was not taken into consideration. It is known that as the duration of therapy increases, the general well-being of ART patients improves ,38 and aSecondly, it is difficult to make conclusions on fertility desires taking only four variables  as determinants. There are other variables like culture, religion, marriage, income, partner influence, and like, which we were not able to make analyses because of either lack of or unfitness of data for meta-analyses. A systematic review by Nattabi et al. in 2009 also concluded that fertility desires are influenced by a myriad of factors  . ThirdlyAlthough the fertility desire among childless and the young group was very strong, in general, we realized that quite a significant segment of HIV-infected people has desires for fertility. The reviewed literature also identified a gap in addressing fertility issue in the routine care for HIV positive people. Therefore, including fertility issue as integral part of HIV patient care and counseling can help several of them in their reproductive decision making. Such counseling needs to focus on letting HIV positive people know the risks and methods of prevention while anticipating a pregnancy.The authors declare that there is no competing interest. For this analysis, we have not got any financial or technical support.YB wrote the whole manuscript and assisted in study selection and data analysis. AB mainly worked on the data analysis. Both authors read and approved the final manuscript.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2458/13/409/prepub