Urate-induced epigenetic modifications in myeloid cells.

OBJECTIVES: Hyperuricemia is a metabolic condition central to gout pathogenesis. Urate exposure primes human monocytes towards a higher capacity to produce and release IL-1ß. In this study, we assessed the epigenetic processes associated to urate-mediated hyper-responsiveness. METHODS: Freshly isolated human peripheral blood mononuclear cells or enriched monocytes were pre-treated with solubilized urate and stimulated with LPS with or without monosodium urate (MSU) crystals. Cytokine production was determined by ELISA. Histone epigenetic marks were assessed by sequencing immunoprecipitated chromatin. Mice were injected intraarticularly with MSU crystals and palmitate after inhibition of uricase and urate administration in the presence or absence of methylthioadenosine. DNA methylation was assessed by methylation array in whole blood of 76 participants with normouricemia or hyperuricemia. RESULTS: High concentrations of urate enhanced the inflammatory response in vitro in human cells and in vivo in mice, and broad-spectrum methylation inhibitors reversed this effect. Assessment of histone 3 lysine 4 trimethylation (H3K4me3) and histone 3 lysine 27 acetylation (H3K27ac) revealed differences in urate-primed monocytes compared to controls. Differentially methylated regions (e.g. HLA-G, IFITM3, PRKAB2) were found in people with hyperuricemia compared to normouricemia in genes relevant for inflammatory cytokine signaling. CONCLUSION: Urate alters the epigenetic landscape in selected human monocytes or whole blood of people with hyperuricemia compared to normouricemia. Both histone modifications and DNA methylation show differences depending on urate exposure. Subject to replication and validation, epigenetic changes in myeloid cells may be a therapeutic target in gout.

The basic morphological characteristics of astrocytomas in Vojvodina in the period 2001-2006.

PURPOSE: Astrocytomas are the most common primary intracranial neoplasms. The aim of this investigation was to register the age, sex, tumor localization, frequency and histological types of patients with astrocytomas. METHODS: The investigation was carried out from January 2001 to June 2006 and included 490 consecutive patients of both sexes with diagnosed intracranial tumors, who had undergone surgical treatment at the Neurosurgery Clinic of the Clinical Centre of Vojvodina. Tumor histological studies were carried out in the Laboratory of the Centre for Pathology and Histology of the Clinical Centre of Vojvodina. Out of 490 patients with diagnosed intracranial tumors, 139 (28.4%) had astrocytomas. RESULTS: Astrocytomas were more frequent in males (63.3%) and were most common in the 50-59-year age group (39.5%). The most common localization was the frontal region (30.2%), more commonly on the right side (51.8%). In regard to other histological types of intracranial tumors, astrocytomas were more frequent in males (34.8%). Grade III astrocytomas were most common (55.4%). The frequency of hemorrhage and thrombosis showed a positive correlation with the histological grade of astrocytomas. CONCLUSION: The typical patient with astrocytoma is a male of 50-59 years. The tumor is grade III located in the right frontal region.

Epidemiology of dietary supplement use in Serbia: Report from Novi Sad.

OBJECTIVES: The primary objectives were to assess the prevalence of dietary supplement (DS) use and to identify specific demographic and lifestyle characteristics of DS users from Novi Sad, Serbia as well as the most commonly used DS and reasons for their use. DESIGN: Observational, cross-sectional study. SETTING AND INTERVENTIONS: Data on demographics, lifestyle and dietary supplement use of 435 adults from Novi Sad, Serbia were collected using an online questionnaire. RESULTS: In total, 435 subjects completed the questionnaire (62.3% women). Prevalence of dietary supplement use in the sample was 42.8%. More women used DS than men (p = 0.002). Higher use of DS was reported among individuals 65+, while the young used DS less (p = 0.001), but the highest proportions of DS users was from the 45-54 age group. DS were used more among those with lower education levels (p < 0.001) and no income (p = 0.009). The highest percentages of DS users reported daily intakes of fruits and moderate physical activity, were non-smokers and social drinkers. Main reason for DS use was maintaining general health. The most commonly used DS were minerals and/or vitamins (68.8%). CONCLUSIONS: We report a high prevalence of dietary supplement use in Novi Sad. DS use was associated with being a female, being older and having minimal/average income, the latter being opposite of the usual findings. Our results warrant a more detailed examination of the association between income, DS use and healthcare availability in developing countries such as Serbia.

Specific tumour-associated methylation in normal human term placenta and first-trimester cytotrophoblasts.

Human placentation displays many similarities with tumourigenesis, including rapid cell division, migration and invasion, overlapping gene expression profiles and escape from immune detection. Recent data have identified promoter methylation in the Ras association factor and adenomatous polyposis coli tumour suppressor genes as part of this process. However, the extent of tumour-associated methylation in the placenta remains unclear. Using whole genome methylation data as a starting point, we have examined this phenomenon in placental tissue. We found no evidence for methylation of the majority of common tumour suppressor genes in term placentas, but identified methylation in several genes previously described in some human tumours. Notably, promoter methylation of four independent negative regulators of Wnt signalling has now been identified in human placental tissue and purified trophoblasts. Methylation is present in baboon, but not in mouse placentas. This supports a role for elevated Wnt signalling in primate trophoblast invasiveness and placentation. Examination of invasive choriocarcinoma cell lines revealed altered methylation patterns consistent with a role of methylation change in gestational trophoblastic disease. This distinct pattern of tumour-associated methylation implicates a coordinated series of epigenetic silencing events, similar to those associated with some tumours, in the distinct features of normal human placental invasion and function.

Methylation of the adenomatous polyposis coli (APC) gene in human placenta and hypermethylation in choriocarcinoma cells.

Methylation of the human APC gene promoter is associated with several different types of cancers and has also been documented in some pre-cancerous tissues. We have examined the methylation of APC gene promoters in human placenta and choriocarcinoma cells. This revealed a general hypomethylation of the APC-1b promoter and a pattern with monoallelic methylation of the APC-1a promoter in full term placental tissue. However, there was no evidence of a parent-of-origin effect, suggesting random post zygotic origin of methylation. Increased methylation of this promoter was observed in all choriocarcinoma-derived trophoblast cell lines, suggesting a trophoblastic origin of placental APC methylation and implicating APC hypermethylation in the development of this group of gestational tumours. Our demonstration of placental methylation of the APC-1a promoter represents the first observation of monoallelic methylation of this gene in early development, and provides further support for a role of canonical Wnt signalling in placental trophoblast invasiveness. This also implicates tumour suppressor gene silencing as an integral part of normal human placental development.

Radiation and scattering from imperfect cylindrical electromagnetic cloaks.

The design of electromagnetic invisibility cloaks is based on singular mappings prescribing zero or infinite values for material parameters on the inner surface of the cloak. Since this is only approximately feasible, an asymptotic analysis is necessary for a sound description of cloaks. We adopt a simple and effective approach for analyzing electromagnetic cloaks - instead of the originally proposed singular mapping, nonsingular mappings asymptotically approaching the ideal one are considered. Scattering and radiation from this type of imperfect cylindrical cloaks is solved analytically and the results are confirmed by full-wave finite element simulations. Our analysis sheds more light on the influence of this kind of imperfection on the cloaking performance and further explores the physics of cloaking devices.

DNA methylation of amino acid transporter genes in the human placenta.

INTRODUCTION: Placental transfer of amino acids via amino acid transporters is essential for fetal growth. Little is known about the epigenetic regulation of amino acid transporters in placenta. This study investigates the DNA methylation status of amino acid transporters and their expression across gestation in human placenta. METHODS: BeWo cells were treated with 5-aza-2'-deoxycytidine to inhibit methylation and assess the effects on amino acid transporter gene expression. The DNA methylation levels of amino acid transporter genes in human placenta were determined across gestation using DNA methylation array data. Placental amino acid transporter gene expression across gestation was also analysed using data from publically available Gene Expression Omnibus data sets. The expression levels of these transporters at term were established using RNA sequencing data. RESULTS: Inhibition of DNA methylation in BeWo cells demonstrated that expression of specific amino acid transporters can be inversely associated with DNA methylation. Amino acid transporters expressed in term placenta generally showed low levels of promoter DNA methylation. Transporters with little or no expression in term placenta tended to be more highly methylated at gene promoter regions. The transporter genes SLC1A2, SLC1A3, SLC1A4, SLC7A5, SLC7A11 and SLC7A10 had significant changes in enhancer DNA methylation across gestation, as well as gene expression changes across gestation. CONCLUSION: This study implicates DNA methylation in the regulation of amino acid transporter gene expression. However, in human placenta, DNA methylation of these genes remains low across gestation and does not always play an obvious role in regulating gene expression, despite clear evidence for differential expression as gestation proceeds.

Increased risk of neuroectodermal tumors and stomach cancer in relatives of patients with Ewing's sarcoma family of tumors.

BACKGROUND: Previous studies of the genetic epidemiology of Ewing's sarcoma have shown neither an increased incidence nor a distinct pattern of cancers in family members of Ewing's sarcoma patients. PURPOSE: Because of a new biologic and cytogenetic classification of Ewing's sarcoma family of tumors, we wanted to reinvestigate the incidence and distribution of cancers in relatives of probands with Ewing's sarcoma family of tumors. METHODS: Patients treated at the Pediatric Branch and the Radiation Oncology Branch of the National Cancer Institute between 1965 and December 1992, or their next of kin, were asked to complete a questionnaire on the history of cancer in all first- and second-degree relatives. The incidence of cancer in family members was compared with Connecticut Tumor Registry rates specific for sex, age, and 5-year calendar-year intervals. Observed/expected (O/E) ratios, 95% confidence intervals (CIs), and tests of homogeneity were calculated. RESULTS: Four thousand six hundred seventy-eight family members with 196,640 person-years at risk entered the analysis. Overall, there was no increased risk of cancer (observed 472; O/E = 0.9; 95% CI = 0.8-1.0). However, several tumor types were found in significant excess. These tumors included stomach cancer (observed 34; O/E = 2.0; 95% CI = 1.4-2.8), melanoma (observed 23; O/E = 1.9; 95% CI = 1.2-2.8), brain tumor (observed 18; O/E = 1.9; 95% CI = 1.1-3.0), and bone cancer (observed 7; O/E = 4.2; 95% CI = 1.7-8.6). Risks of these cancers were higher among maternal than paternal relatives, but these differences were not statistically significant. There was a significant deficit of bladder cancer (observed 5; O/E = 0.2; 95% CI = 0.1-0.5) and rectal cancer (observed 0; O/E = 0.0; 95% CI = 0.0-0.1). Second-degree relatives had a significant cancer deficit (observed 389; O/E = 0.9; 95% CI = 0.8-0.95). This deficit was accounted for by the observed deficit of bladder and rectal cancer and is probably related to under-reporting or misclassification of cancer in second-degree relatives. Family members of 10 probands with second malignancies did not have an increased risk of all cancers (observed 20; O/E = 1.2; 95% CI = 0.7-1.8) but had an increased risk of both melanoma (observed 3; O/E = 7.3; 95% CI = 1.5-21.0) and breast cancer (observed 8; O/E = 3.2; 95% CI = 1.4-6.3). CONCLUSION: Finding an increased risk of neuroectodermal tumors and stomach cancer in families of patients with Ewing's sarcoma family of tumors suggests that these tumors might share a common etiology. Further studies should try to confirm this hypothesis and to examine if genetic factors may have a role in these families by assessing the mode of inheritance and examining families with multiple affected members.

The effects of maternal anxiety during pregnancy on IGF2/H19 methylation in cord blood.

Compelling evidence suggests that maternal mental health in pregnancy can influence fetal development. The imprinted genes, insulin-like growth factor 2 (IGF2) and H19, are involved in fetal growth and each is regulated by DNA methylation. This study aimed to determine the association between maternal mental well-being during pregnancy and differentially methylated regions (DMRs) of IGF2 (DMR0) and the IGF2/H19 imprinting control region (ICR) in newborn offspring. Maternal depression, anxiety and perceived stress were assessed at 28 weeks of pregnancy in the Barwon Infant Study (n=576). DNA methylation was measured in purified cord blood mononuclear cells using the Sequenom MassArray Platform. Maternal anxiety was associated with a decrease in average ICR methylation (Δ=-2.23%; 95% CI=-3.68 to -0.77%), and across all six of the individual CpG units in anxious compared with non-anxious groups. Birth weight and sex modified the association between prenatal anxiety and infant methylation. When stratified into lower (⩽3530 g) and higher (>3530 g) birth weight groups using the median birth weight, there was a stronger association between anxiety and ICR methylation in the lower birth weight group (Δ=-3.89%; 95% CI=-6.06 to -1.72%), with no association in the higher birth weight group. When stratified by infant sex, there was a stronger association in female infants (Δ=-3.70%; 95% CI=-5.90 to -1.51%) and no association in males. All the linear regression models were adjusted for maternal age, smoking and folate intake. These findings show that maternal anxiety in pregnancy is associated with decreased IGF2/H19 ICR DNA methylation in progeny at birth, particularly in female, low birth weight neonates. ICR methylation may help link poor maternal mental health and adverse birth outcomes, but further investigation is needed.

IFPA meeting 2014 workshop report: Animal models to study pregnancy pathologies; new approaches to study human placental exposure to xenobiotics; biomarkers of pregnancy pathologies; placental genetics and epigenetics; the placenta and stillbirth and fetal growth restriction.

Workshops are an important part of the IFPA annual meeting as they allow for discussion of specialized topics. At IFPA meeting 2014 there were six themed workshops, five of which are summarized in this report. These workshops related to various aspects of placental biology but collectively covered areas of animal models, xenobiotics, pathological biomarkers, genetics and epigenetics, and stillbirth and fetal growth restriction.

Can protocol-specified doses of chemotherapy and radiotherapy be used as a measure of treatment actually received? A CCG/NIH study on long-term survivors of acute lymphocytic leukemia.

In a cohort of 593 long-term survivors of acute lymphocytic leukemia identified through the Children's Cancer Group (CCG), treatment abstracts were obtained and compared to protocol information on radiation therapy and intravenous chemotherapy. This was done in order to evaluate the actual compliance to protocol-specified treatment, and assess if protocol-specified doses can be used in studies of late effects of treatment. The compliance to protocol-specified type of treatment ranged between 95.3% (intrathecal methotrexate) and 98.6% (adriamycin) for chemotherapy, and between 94.1% (cranial radiation) and 97.0% (extended field radiation) for radiation. Concordance with the protocol-specified chemotherapy dose (+/- 25%) was 57.5% for adriamycin, 91.3% for daunomycin, and 48.5% for cyclophosphamide. When concordance was low, most patients received doses that were lower than expected. Concordance with chemotherapy was significantly lower for high-dose regimens than for low-dose regimens. Concordance with protocol-specified radiation dose (+/- 10%) was 87.4% for cranial radiation, 87.8% for spinal radiation, and 85.7% for extended field radiation. Concordance with treatment did not differ by gender, relapse status, or age at diagnosis. In this cohort of leukemia survivors, the validity of type of treatment was greater than the validity of dosage. Great care should be used when drawing conclusions about effects of treatment dosage. Although costly and time consuming, it appears that chart reviews are the most appropriate way to collect information about dose-related effects of therapy.

Absorbed fraction of radon progeny in human bronchial airways with bifurcation geometry.

PURPOSE: The absorbed fraction, defined as the portion of the initial particle energy which is absorbed in the tissue of interest, was calculated, under bifurcation geometry of the airway tubes, for alpha-particles emitted from radon progeny in the human respiratory tract. The results are given for all branching generations and compared with the data obtained for the commonly used infinite straight cylinders adopted by the International Commission on Radiological Protection (ICRP) Report 66. MATERIALS AND METHODS: A model was created to calculate the absorbed fraction of alpha-particle energy in the human lung using bifurcation geometry. Monte Carlo simulations of alpha-particle propagation in tissue and air were performed. The stopping powers of alpha-particles were adopted from the International Commission of Radiation Units and Measurements (ICRU) Report 49. RESULTS: The absorbed fractions for the bifurcation geometry are given for the 15 generations in the tracheobronchial tree for alpha-particle energies of 6 and 7.69 MeV. The sources were assumed to be the fast and slow moving mucus. CONCLUSIONS: Comparisons with ICRP66 data reveal that the assumption of long, straight cylinders was appropriate in some cases, but not in all. Adoption of the absorbed fractions obtained from the bifurcation model instead of the ICRP66 data caused 'redistribution' of doses in the bronchial (BB) and bronchiolar (bb) regions.

The immunohistochemical and serological determination of p53 protein in patients with malignant lymphomas.

The product of mutated p53 gene is a protein with abnormal conformation, impaired DNA binding, and a prolonged half life, the latter of which results in immunohistochemically detectable levels within nuclei of malignant cells. The present study was aimed at the immunohistochemical determination of p53 overexpression in patients with various histological types of nonHodgkin's lymphomas (NHL), with a particular interest in gastric lymphomas. In these patients, as well as in controls, also serological determinations of p53 protein were performed using an ELISA method. Immunohistochemical overexpression of p53 protein was found in 21% of NHL patients, with the highest incidence of p53 immunoreactivity in cases of Burkitt's lymphoma, follicle center lymphoma grade III, and diffuse large B-cell lymphoma. In gastric lymphomas the overall incidence of p53 immunoreactivity was as high as 46%. Serological ELISA determinations of p53 protein in NHL patients and in controls remained below the lowest detection limit of the method in all 128 cases. Considering that p53 mutations are associated with poor response to therapy, and consequently with poor prognosis, it is of great importance to determine the subset of patients that are particularly at risk for an unfavorable outcome and should be treated more aggressively. Immunohistochemical determinations of p53 overexpression represent a rapid and simple, yet somewhat imperfect technique for an estimation of the frequency of mutational events. On the other hand, serological determinations of p53 protein are completely inadequate for the evaluation of p53 status.

Telomerase activity as a biological marker in some gynecological tumors: comparison with tissue and serum CA 125.

BACKGROUND: Since telomerase activity is detectable in cancer cells but not in some normal somatic cells, it has been considered as a potential diagnostic marker as well as a target for possible anticancer strategies. The purpose of this study was to assess the value of telomerase activity determination in some gynecological tumors and to compare it with the CA 125 tissue and serum profile. PATIENTS AND METHODS: The telomerase activity was determined in 11 gynecological tumors: 7 ovarian carcinomas, 2 carcinomas of the fallopian tube and 2 cervical carcinomas, and compared to the activity in the normal peritoneal tissue of the same patients. Additionally, the levels of CA 125 were measured in the tumor and normal peritoneum tissue samples as well as in the patients' sera. RESULTS: In ovarian tumors, the telomerase activity was detected in 71.4% (5 out of 7), while in the carcinomas of the fallopian tube and cervical carcinomas such activity was not observed. Negative for telomerase activity were also all samples of peritoneum. The range of CA 125 in the tumor tissue was 99 U/g-803667 U/g of tissue and in the normal peritoneum 71 U/g-4925 U/g of tissue. CONCLUSION: In conclusion, telomerase activity could be detected in some of the gynecological tumors, but for clinical use as a diagnostic marker it is inferior to CA 125.

Experiences of cancer in children and adolescents.

In an effort to understand the effect of cancer diagnosis and treatment in children and adolescents, and to identify issues that should be addressed with newly diagnosed patients, 85 patients with Ewing's sarcoma family tumors (ESFT) were interviewed about their experience of having cancer. This represents 90% of all eligible patients who survived at least 3 years since their diagnosis and who were treated for ESFT at the National Cancer Institute (NCI) from 1965-1993. The mean age of patients at the time of diagnosis was 15.8 +/- 5.3 years, and mean time since diagnosis was 13.6 +/- 6.4 years. Patients from this cohort had a disease usually related to poor outcome. Patients answered five open-ended written questions. Negative experiences that they described included transient and permanent discomfort and disabilities related to cancer; disruption of life or relationships; and emotional aspects of cancer diagnosis or treatment. Positive aspects of having cancer included changed attitudes about self and life, improved relationships with others, or better job performance. Advice for newly diagnosed patients most often dealt with the emotional aspects of cancer. The importance of patient-to-patient support was frequently described. Overall, having cancer was not an entirely negative experience, and it may result in introspection and improved relationships with others.

Discrete time neural network synthesis using input and output activation functions.

A new very fast algorithm for synthesis of a new structure of discrete-time neural networks (NN) is proposed. For this purpose the following concepts are employed: (i) combination of input and output activation functions, (ii) input time-varying signal distribution, (iii) time-discrete domain synthesis and (iv) one-step learning iteration approach. The problem of input-output mappings of time-varying vectors is solved. Simulation results based on the synthesis of a new structure of feedforward NN of an universal logical unit are presented. The proposed NN synthesis procedure is useful for applications to identification and control of nonlinear, very fast, dynamical systems. In this sense a feedforward NN for an adaptive nonlinear robot control is designed. Finally, a new algorithm for the direct inverse modeling of input/output nonquadratic systems is discussed.

Fuzzy logic control synthesis without any rule base.

A new analytic fuzzy logic control (FLC) system synthesis without any rule base is proposed. For this purpose the following objectives are preferred and reached: 1) an introduction of a new adaptive shape of fuzzy sets and a new adaptive distribution of input fuzzy sets, 2) a determination of an analytic activation function for activation of output fuzzy sets, instead of using of min-max operators, and 3) a definition of a new analytic function that determines the positions of centers of output fuzzy sets in each mapping process, instead of definition of the rule base. A real capability of the proposed FLC synthesis procedures is presented by synthesis of FLC of robot of RRTR-structure.

The ever growing complexity of placental epigenetics - role in adverse pregnancy outcomes and fetal programming.

As the primary interface between maternal and fetal circulations, the placenta is subject to a myriad of environmental exposures with the capacity to alter placental function and fetal development. Many of these effects are likely to be mediated by epigenetic ('above DNA') change, which is also in turn regulated by maternal and fetal genetic factors. Linking specific environmental exposures, genetic, and epigenetic variation to maternal and fetal outcomes may provide valuable mechanistic insights into the role of placental dysfunction in pregnancy-associated disease and later health. The complexities are manifold but are rapidly being overcome by technological advances and emerging analytical approaches. Although focussing on recent genome-scale and gene-specific DNA methylation studies in the human placenta, this review also discusses the potential of a future broader exploration of combined environmental, genetic and epigenomic approaches, encompassing higher order epigenetic modifications, for unravelling the molecular mechanisms underlying gene-environment interaction at the fetomaternal interface.

IFPA Meeting 2010 Workshop Report I: Immunology; ion transport; epigenetics; vascular reactivity; epitheliochorial placentation; proteomics.

Workshops are an important part of the IFPA annual meeting. At IFPA Meeting 2010 there were twelve themed workshops, six of which are summarized in this report. 1. The immunology workshop focused on normal and pathological functions of the maternal immune system in pregnancy. 2. The transport workshop dealt with regulation of ion and water transport across the syncytiotrophoblast of human placenta. 3. The epigenetics workshop covered DNA methylation and its potential role in regulating gene expression in placental development and disease. 4. The vascular reactivity workshop concentrated on methodological approaches used to study placental vascular function. 5. The workshop on epitheliochorial placentation covered current advances from in vivo and in vitro studies of different domestic species. 6. The proteomics workshop focused on a variety of techniques and procedures necessary for proteomic analysis and how they may be implemented for placental research.