Mathematical model of a telomerase transcriptional regulatory network developed by cell-based screening: analysis of inhibitor effects and telomerase expression mechanisms

Cancer cells depend on transcription of telomerase reverse transcriptase (TERT). Many transcription factors affect TERT, though regulation occurs in context of a broader network. Network effects on telomerase regulation have not been investigated, though deeper understanding of TERT transcription requires a systems view. However, control over individual interactions in complex networks is not easily achievable. Mathematical modelling provides an attractive approach for analysis of complex systems and some models may prove useful in systems pharmacology approaches to drug discovery. In this report, we used transfection screening to test interactions among 14 TERT regulatory transcription factors and their respective promoters in ovarian cancer cells. The results were used to generate a network model of TERT transcription and to implement a dynamic Boolean model whose steady states were analysed. Modelled effects of signal transduction inhibitors successfully predicted TERT repression by Src-family inhibitor SU6656 and lack of repression by ERK inhibitor FR180204, results confirmed by RT-QPCR analysis of endogenous TERT expression in treated cells. Modelled effects of GSK3 inhibitor 6-bromoindirubin-3′-oxime (BIO) predicted unstable TERT repression dependent on noise and expression of JUN, corresponding with observations from a previous study. MYC expression is critical in TERT activation in the model, consistent with its well known function in endogenous TERT regulation. Loss of MYC caused complete TERT suppression in our model, substantially rescued only by co-suppression of AR. Interestingly expression was easily rescued under modelled Ets-factor gain of function, as occurs in TERT promoter mutation. RNAi targeting AR, JUN, MXD1, SP3, or TP53, showed that AR suppression does rescue endogenous TERT expression following MYC knockdown in these cells and SP3 or TP53 siRNA also cause partial recovery. The model therefore successfully predicted several aspects of TERT regulation including previously unknown mechanisms. An extrapolation suggests that a dominant stimulatory system may programme TERT for transcriptional stability

Development of a cDNA microarray for the measurement of gene expression in the sheep scab mite Psoroptes ovis

Background: Sheep scab is caused by the ectoparasitic mite Psoroptes ovis which initiates a profound cutaneous inflammatory response, leading to the development of the skin lesions which are characteristic of the disease. Existing control strategies rely upon injectable endectocides and acaricidal dips but concerns over residues, eco-toxicity and the development of acaricide resistance limit the sustainability of this approach. In order to identify alternative means of disease control, a deeper understanding of both the parasite and its interaction with the host are required. Methods: Herein we describe the development and utilisation of an annotated P. ovis cDNA microarray containing 3,456 elements for the measurement of gene expression in this economically important ectoparasite. The array consists of 981 P. ovis EST sequences printed in triplicate along with 513 control elements. Array performance was validated through the analysis of gene expression differences between fed and starved P. ovis mites. Results: Sequences represented on the array include homologues of major house dust mite allergens and tick salivary proteins, along with factors potentially involved in mite reproduction and xenobiotic metabolism. In order to validate the performance of this unique resource under biological conditions we used the array to analyse gene expression differences between fed and starved P. ovis mites. These analyses identified a number of house dust mite allergen homologues up-regulated in fed mites and P. ovis transcripts involved in stress responses, autophagy and chemosensory perception up-regulated in starved mites. Conclusion: The P. ovis cDNA microarray described here has been shown to be both robust and reproducible and will enable future studies to analyse gene expression in this important ectoparasite

Epidemiology of multimorbidity within the Brazilian adult general population:Evidence from the 2013 National Health Survey (PNS 2013)

Middle-income countries are facing a growing challenge of adequate health care provision for people with multimorbidity. The objectives of this study were to explore the distribution of multimorbidity and to identify patterns of multimorbidity in the Brazilian general adult population. Data from 60202 adults, aged ≥18 years that completed the individual questionnaire of the National Health Survey 2013 (Portuguese: "Pesquisa Nacional de Saúde"-"PNS") was used. We defined multimorbidity as the presence of two or more chronic conditions, including self-reported diagnoses and responses to the 9-item Patient Health Questionnaire for depression. Multivariate Poisson regression analyses were used to explore relationship between multimorbidity and demographic factors. Exploratory tetrachoric factor analysis was performed to identify multimorbidity patterns. 24.2% (95% CI 23.5-24.9) of the study population were multimorbid, with prevalence rate ratios being significantly higher in women, older people and those with lowest educational level. Multimorbidity occurred earlier in women than in men, with half of the women and men aged 55-59 years and 65-69 years, respectively, were multimorbid. The absolute number of people with multimorbidity was approximately 2.5-fold higher in people younger than 65 years than older counterparts (9920 vs 3945). Prevalence rate ratios of any mental health disorder significantly increased with the number of physical conditions. 46.7% of the persons were assigned to at least one of three identified patterns of multimorbidity, including: "cardio-metabolic", "musculoskeletal-mental" and "respiratory" disorders. Multimorbidity in Brazil is as common as in more affluent countries. Women in Brazil develop diseases at younger ages than men. Our findings can inform a national action plan to prevent multimorbidity, reduce its burden and align health-care services more closely with patients' needs