Towards understanding the gliotoxin detoxification mechanism: in vivo thiomethylation protects yeast from gliotoxin cytotoxicity

Gliotoxin (GT) is a mycotoxin produced by some species of ascomycete fungi including the opportunistic human pathogen Aspergillus fumigatus . In order to produce GT the host organism needs to have evolved a selfprotection mechanism. GT contains a redox-cycling disulfide bridge that is important in mediating toxicity. Recently is has been demonstrated that A. fumigatus possesses a novel thiomethyltransferase protein called GtmA that has the ability to thiomethylate GT in vivo , which aids the organism in regulating GT biosynthesis. It has been suggested that thiomethylation of GT and similar sulfur-containing toxins may play a role in providing self-protection in host organisms. In this work we have engineered Saccharomyces cerevisiae , a GT-naïve organism, to express A. fumigatus GtmA. We demonstrate that GtmA can readily thiomethylate GT in yeast, which results in protection of the organism from exogenous GT. Our work has implications for understanding the evolution of GT self-protection mechanisms in organisms that are GT producers and non-producers

The long-term impact of the MEMA kwa Vijana adolescent sexual and reproductive health intervention: effect of dose and time since intervention exposure.

BACKGROUND: Despite recent decreases in HIV incidence in many sub-Saharan African countries, there is little evidence that specific behavioural interventions have led to a reduction in HIV among young people. Further and wider-scale decreases in HIV require better understanding of when behaviour change occurs and why. The MEMA kwa Vijana adolescent sexual and reproductive health intervention has been implemented in rural Mwanza, Tanzania since 1999. A long-term evaluation in 2007/8 found that the intervention improved knowledge, attitudes to sex and some reported risk behaviours, but not HIV or HSV2 prevalence. The aim of this paper was to assess the differential impact of the intervention according to gender, age, marital status, number of years of exposure and time since last exposure to the intervention. METHODS: In 2007, a cross-sectional survey was conducted in the 20 trial communities among 13,814 young people (15-30 yrs) who had attended intervention or comparison schools between 1999 and 2002. Outcomes for which the intervention had an impact in 2001 or 2007 were included in this subgroup analysis. Data were analysed using cluster-level methods for stratified cluster-randomised trials, using interaction tests to determine if intervention impact differed by subgroup. RESULTS: Taking into account multiplicity of testing, concurrence with a priori hypotheses and consistency within the results no strong effect-modifiers emerged. Impact on pregnancy knowledge and reported attitudes to sex increased with years of exposure to high-quality intervention. CONCLUSIONS: The desirable long-term impact of the MEMA kwa Vijana intervention did not vary greatly according to the subgroups examined. This suggests that the intervention can have an impact on a broad cross-section of young people in rural Mwanza. TRIAL REGISTRATION: ClinicalTrials.gov NCT00248469

RNA-seq reveals the pan-transcriptomic impact of attenuating the gliotoxin self-protection mechanism in Aspergillus fumigatus.

BACKGROUND: Aspergillus fumigatus produces a number of secondary metabolites, one of which, gliotoxin, has been shown to exhibit anti-fungal activity. Thus, A. fumigatus must be able to protect itself against gliotoxin. Indeed one of the genes in the gliotoxin biosynthetic gene cluster in A. fumigatus, gliT, is required for self-protection against the toxin- however the global self-protection mechanism deployed is unclear. RNA-seq was employed to identify genes differentially regulated upon exposure to gliotoxin in A. fumigatus wild-type and A. fumigatus ∆gliT, a strain that is hypersensitive to gliotoxin. RESULTS: Deletion of A. fumigatus gliT resulted in altered expression of 208 genes (log2 fold change of 1.5) when compared to A. fumigatus wild-type, of which 175 genes were up-regulated and 33 genes were down-regulated. Expression of 164 genes was differentially regulated (log2 fold change of 1.5) in A. fumigatus wild-type when exposed to gliotoxin, consisting of 101 genes with up-regulated expression and 63 genes with down-regulated expression. Interestingly, a much larger number of genes, 1700, were found to be differentially regulated (log2 fold change of 1.5) in A. fumigatus ∆gliT when challenged with gliotoxin. These consisted of 508 genes with up-regulated expression, and 1192 genes with down-regulated expression. Functional Catalogue (FunCat) classification of differentially regulated genes revealed an enrichment of genes involved in both primary metabolic functions and secondary metabolism. Specifically, genes involved in gliotoxin biosynthesis, helvolic acid biosynthesis, siderophore-iron transport genes and also nitrogen metabolism genes and ribosome biogenesis genes underwent altered expression. It was confirmed that gliotoxin biosynthesis is induced upon exposure to exogenous gliotoxin, production of unrelated secondary metabolites is attenuated in A. fumigatus ∆gliT, while quantitative proteomic analysis confirmed disrupted translation in A. fumigatus ∆gliT challenged with exogenous gliotoxin. CONCLUSIONS: This study presents the first global investigation of the transcriptional response to exogenous gliotoxin in A. fumigatus wild-type and the hyper-sensitive strain, ∆gliT. Our data highlight the global and extensive affects of exogenous gliotoxin on a sensitive strain devoid of a self-protection mechanism and infer that GliT functionality is required for the optimal biosynthesis of selected secondary metabolites in A. fumigatus

Aplicación de modelos bayesianos para estimar la prevalencia de enfermedad y la sensibilidad y especificidad de tests de diagnóstico clínico sin gold standard

[spa] Dos objetivos claves de la investigación diagnóstica son estimar con exactitud y precisión la prevalencia de la enfermedad y la sensibilidad y especificidad de tests diagnósticos. Se han desarrollado modelos de clases latentes que tienen en cuenta la correlación entre las medidas de los individuos determinadas con diferentes tests con el fin de diagnosticar enfermedades para las cuales no están disponibles tests gold standard. En algunos estudios clínicos, se hacen varias medidas del mismo individuo con el mismo test en las mismas condiciones y, por tanto, las mediciones replicadas para cada individuo no son independientes. En esta tesis se propone una extensión Bayesiana del modelo de clases latentes de efectos aleatorios de Gauss para ajustar a los datos de tests con resultados binarios y con medidas replicadas por individuo. Se describe una aplicación que utiliza los datos recogidos de personas infectadas por parásitos intestinales Hookworm llevada a cabo en el municipio de Presidente Figueiredo, Estado de Amazonas en Brasil. Además, a través de un estudio de simulación se comparó el desempeño del modelo propuesto con los modelos actuales (el modelo de efectos aleatorios individuo y modelos de dependencia e independencia condicional). Como era de esperar, el modelo propuesto presenta una mayor exactitud y precisión en las estimaciones de prevalencia, sensibilidad y especificidad. Para un control adecuado de las enfermedades la Organización Mundial de la Salud ha propuesto el diagnóstico y tratamiento de la infección tuberculosa latente (LTBI) en grupos de riesgo de desarrollar la enfermedad, como los niños. No existe un test gold standard para el diagnóstico de la infección latente. Los modelos estadísticos basados en la estimación de clases latentes permiten la evaluación de la prevalencia de la infección y la validez de los tests utilizados en ausencia de un gold standard. Se realizó un estudio transversal con niños de hasta 6 años de edad que habían sido vacunados con la BCG en Manaus, Amazonas-Brasil. El objetivo de dicho estudio fue estimar la prevalencia de la infección latente en los niños pequeños en contacto con un caso indice de tuberculosis en el hogar (TB-HCC) y determinar la validez y la seguridad del test cutáneo de tuberculina (TST) y QuantiFERON-TB Gold-in-tube (QFT), utilizando modelos de clases latentes. Para las estimaciones, en una primera fase se consideró la correlación entre los dos tests, y en la segunda fase se consideró la prevalencia en función de la intensidad y del tiempo de exposición. El cincuenta por ciento de los niños con TB-HCC tenía LTBI, con la prevalencia en función del tiempo y la intensidad de la exposición al caso índice. La sensibilidad y la especificidad de TST fueron del 73 % [intervalo de confianza del 95 % (IC): 53-91] y el 97 % (IC del 95 %: 89-100), respectivamente, frente al 53 % (IC del 95 %: 41-66) y el 81 % (IC del 95 %: 71-90) para QFT. El valor predictivo positivo de TST en niños con TB-HCC fue del 91 % (IC del 95 %: 61-99), y para QFT fue del 74 % (IC del 95 %: 47-95). Este es uno de los primeros estudios que usa modelos de clases latentes para estimar la prevalencia de la infección por M. tuberculosis en niños y los parámetros de sus principales tests diagnósticos. Los resultados sugieren que los niños en contacto con un caso índice tienen un alto riesgo de infección. La validez y los valores predictivos no mostraron diferencias significativas según el test aplicado. El uso combinado de los dos tests en nuestro estudio mostró una sutil mejoría en el diagnóstico de la LTBI.[eng] Two key aims of diagnostic research are to accurately and precisely estimate disease prevalence and test sensitivity and specificity. Latent class models have been proposed that consider the correlation between subject measures determined by different tests in order to diagnose diseases for which gold standard tests are not available. In some clinical studies, several measures of the same subject are made with the same test under the same conditions (replicated measurements) and thus, replicated measurements for each subject are not independent. In the present study, we propose an extension of the Bayesian latent class Gaussian random effects model to fit the data with binary outcomes for tests with replicated subject measures. We describe an application using data collected on hookworm infection carried out in the municipality of Presidente Figueiredo, Amazonas State, Brazil. In addition, the performance of the proposed model was compared with that of current models (the subject random effects model and the conditional (in)dependent model) through a simulation study. As expected, the proposed model presented better accuracy and precision in the estimations of prevalence, sensitivity and specificity. For adequate disease control the World Health Organization has proposed the diagnosis and treatment of latent tuberculous infection (LTBI) in groups of risk of developing the disease such as children. There is no gold standard (GS) test for the diagnosis of LTBI. Statistical models based on the estimation of latent class allow evaluation of the prevalence of infection and the accuracy of the tests used in the absence of a GS. We conducted a cross-sectional study with children up to 6 years of age who had been vaccinated with the BCG in Manaus, Amazonas- Brazil. The objective of this study was to estimate the prevalence of LTBI in young children in contact with a household case of tuberculosis (TB-HCC) and determine the accuracy and precision of the Tuberculin Skin Test (TST) and QuantiFERON-TB Gold in-tube (QFT) using the latent class model. Fifty percent of the children with TB-HCC had LTBI, with the pre- valence depending on the intensity and length of exposure to the index case. The sensitivity and specificity of TST were 73 % [95 % confidence interval (CI): 53-91] and 97 % (95 % CI: 89-100), respectively, versus 53 % (95 % CI: 41-66) and 81 % (95 % CI: 71-90) for QFT. The positive predictive value of TST in children with TB-HCC was 91 % (95 % CI: 61-99), and for QFT was 74 % (95 % CI: 47-95). This is one of the first studies to estimate the prevalence of M. tuberculosis infection in children and the parameters of its main diagnostic tests by latent class model. The results suggest that children in contact with an index case have a high risk of infection. The accuracy and the predictive values did not show significant differences according to the test applied. Combined use of the two tests in our study showed scarce improvement in the diagnosis of LTBI

Aplicación de modelos bayesianos para estimar la prevalencia de enfermedad y la sensibilidad y especificidad de tests de diagnóstico clínico sin gold standard

Dos objetivos claves de la investigación diagnóstica son estimar con exactitud y precisión la prevalencia de la enfermedad y la sensibilidad y especificidad de tests diagnósticos. Se han desarrollado modelos de clases latentes que tienen en cuenta la correlación entre las medidas de los individuos determinadas con diferentes tests con el fin de diagnosticar enfermedades para las cuales no están disponibles tests gold standard. En algunos estudios clínicos, se hacen varias medidas del mismo individuo con el mismo test en las mismas condiciones y, por tanto, las mediciones replicadas para cada individuo no son independientes. En esta tesis se propone una extensión Bayesiana del modelo de clases latentes de efectos aleatorios de Gauss para ajustar a los datos de tests con resultados binarios y con medidas replicadas por individuo. Se describe una aplicación que utiliza los datos recogidos de personas infectadas por parásitos intestinales Hookworm llevada a cabo en el municipio de Presidente Figueiredo, Estado de Amazonas en Brasil. Además, a través de un estudio de simulación se comparó el desempeño del modelo propuesto con los modelos actuales (el modelo de efectos aleatorios individuo y modelos de dependencia e independencia condicional). Como era de esperar, el modelo propuesto presenta una mayor exactitud y precisión en las estimaciones de prevalencia, sensibilidad y especificidad. Para un control adecuado de las enfermedades la Organización Mundial de la Salud ha propuesto el diagnóstico y tratamiento de la infección tuberculosa latente (LTBI) en grupos de riesgo de desarrollar la enfermedad, como los niños. No existe un test gold standard para el diagnóstico de la infección latente. Los modelos estadísticos basados en la estimación de clases latentes permiten la evaluación de la prevalencia de la infección y la validez de los tests utilizados en ausencia de un gold standard. Se realizó un estudio transversal con niños de hasta 6 años de edad que habían sido vacunados con la BCG en Manaus, Amazonas-Brasil. El objetivo de dicho estudio fue estimar la prevalencia de la infección latente en los niños pequeños en contacto con un caso indice de tuberculosis en el hogar (TB-HCC) y determinar la validez y la seguridad del test cutáneo de tuberculina (TST) y QuantiFERON-TB Gold-in-tube (QFT), utilizando modelos de clases latentes. Para las estimaciones, en una primera fase se consideró la correlación entre los dos tests, y en la segunda fase se consideró la prevalencia en función de la intensidad y del tiempo de exposición. El cincuenta por ciento de los niños con TB-HCC tenía LTBI, con la prevalencia en función del tiempo y la intensidad de la exposición al caso índice. La sensibilidad y la especificidad de TST fueron del 73 % [intervalo de confianza del 95 % (IC): 53-91] y el 97 % (IC del 95 %: 89-100), respectivamente, frente al 53 % (IC del 95 %: 41-66) y el 81 % (IC del 95 %: 71-90) para QFT. El valor predictivo positivo de TST en niños con TB-HCC fue del 91 % (IC del 95 %: 61-99), y para QFT fue del 74 % (IC del 95 %: 47-95). Este es uno de los primeros estudios que usa modelos de clases latentes para estimar la prevalencia de la infección por M. tuberculosis en niños y los parámetros de sus principales tests diagnósticos. Los resultados sugieren que los niños en contacto con un caso índice tienen un alto riesgo de infección. La validez y los valores predictivos no mostraron diferencias significativas según el test aplicado. El uso combinado de los dos tests en nuestro estudio mostró una sutil mejoría en el diagnóstico de la LTBI.Two key aims of diagnostic research are to accurately and precisely estimate disease prevalence and test sensitivity and specificity. Latent class models have been proposed that consider the correlation between subject measures determined by different tests in order to diagnose diseases for which gold standard tests are not available. In some clinical studies, several measures of the same subject are made with the same test under the same conditions (replicated measurements) and thus, replicated measurements for each subject are not independent. In the present study, we propose an extension of the Bayesian latent class Gaussian random effects model to fit the data with binary outcomes for tests with replicated subject measures. We describe an application using data collected on hookworm infection carried out in the municipality of Presidente Figueiredo, Amazonas State, Brazil. In addition, the performance of the proposed model was compared with that of current models (the subject random effects model and the conditional (in)dependent model) through a simulation study. As expected, the proposed model presented better accuracy and precision in the estimations of prevalence, sensitivity and specificity. For adequate disease control the World Health Organization has proposed the diagnosis and treatment of latent tuberculous infection (LTBI) in groups of risk of developing the disease such as children. There is no gold standard (GS) test for the diagnosis of LTBI. Statistical models based on the estimation of latent class allow evaluation of the prevalence of infection and the accuracy of the tests used in the absence of a GS. We conducted a cross-sectional study with children up to 6 years of age who had been vaccinated with the BCG in Manaus, Amazonas- Brazil. The objective of this study was to estimate the prevalence of LTBI in young children in contact with a household case of tuberculosis (TB-HCC) and determine the accuracy and precision of the Tuberculin Skin Test (TST) and QuantiFERON-TB Gold in-tube (QFT) using the latent class model. Fifty percent of the children with TB-HCC had LTBI, with the pre- valence depending on the intensity and length of exposure to the index case. The sensitivity and specificity of TST were 73 % [95 % confidence interval (CI): 53-91] and 97 % (95 % CI: 89-100), respectively, versus 53 % (95 % CI: 41-66) and 81 % (95 % CI: 71-90) for QFT. The positive predictive value of TST in children with TB-HCC was 91 % (95 % CI: 61-99), and for QFT was 74 % (95 % CI: 47-95). This is one of the first studies to estimate the prevalence of M. tuberculosis infection in children and the parameters of its main diagnostic tests by latent class model. The results suggest that children in contact with an index case have a high risk of infection. The accuracy and the predictive values did not show significant differences according to the test applied. Combined use of the two tests in our study showed scarce improvement in the diagnosis of LTBI

Native geometry and the dynamics of protein folding

In this paper we investigate the role of native geometry on the kinetics of protein folding based on simple lattice models and Monte Carlo simulations. Results obtained within the scope of the Miyazawa-Jernigan indicate the existence of two dynamical folding regimes depending on the protein chain length. For chains larger than 80 amino acids the folding performance is sensitive to the native state's conformation. Smaller chains, with less than 80 amino acids, fold via two-state kinetics and exhibit a significant correlation between the contact order parameter and the logarithmic folding times. In particular, chains with N=48 amino acids were found to belong to two broad classes of folding, characterized by different cooperativity, depending on the contact order parameter. Preliminary results based on the G\={o} model show that the effect of long range contact interaction strength in the folding kinetics is largely dependent on the native state's geometry.Comment: Proceedings of the BIFI 2004 - I International Conference, Zaragoza (Spain) Biology after the genome: a physical view. To appear in Biophysical Chemistr

Mapania multiflora, a distinctive new species of Cyperaceae (Mapanioideae) from Borneo

Mapania multiflora is described and illustrated. It is vegetatively similar to taxa with broad leaves and pseudopetioles, such as M. cuspidata. However, it is reproductively similar to sect. Thoractostachyum with a paniculate inflorescence and furrowed fruit. The DNA is similar to M. bancana in sect. Thoractostachyum, in the three sampled cpDNA regions: atpH-F, trnL-F and psbA-trnH. However, it is identical to none of these due to its unique combination of vegetative, reproductive and molecular characteristics

Interleukin-33 regulates tissue remodelling and inhibits angiogenesis in the eye

Age-related macular degeneration (AMD) is the leading cause of central vision loss worldwide. Loss of retinal pigment epithelium (RPE) is a major pathological hallmark in AMD with or without pathological neovascularization. Although activation of the immune system is implicated in disease progression, pathological pathways remain diverse and unclear. Here, we report an unexpected protective role of a pro-inflammatory cytokine, interleukin-33 (IL-33) in ocular angiogenesis. IL-33 and its receptor (ST2) are expressed constitutively in human and murine retina and choroid. When RPE was activated, IL-33 expression was markedly elevated in vitro. We found that IL-33 regulated tissue remodelling by attenuating wound-healing responses, including reduction in migration of choroidal fibroblasts and retinal microvascular endothelial cells, and inhibition of collagen gel contraction. In vivo, local administration of recombinant IL-33 inhibited murine choroidal neovascularization (CNV) formation, a surrogate of human neovascular AMD, and this effect was ST2-dependent. Collectively, these data demonstrate IL-33 as a potential immunotherapy and distinguishes pathways for subverting AMD pathology

Where to for Sexual Health Education for Adolescents in Sub-Saharan Africa?

Rachel Jewkes discusses disappointing results from a school-based sexual health intervention study in Tanzania and their implications for future health education programs

Outer boundary conditions for evolving cool white dwarfs

White dwarf evolution is essentially a gravothermal cooling process, which,for cool white dwarfs, sensitively depends on the treatment of the outer boundary conditions. We provide detailed outer boundary conditions appropriate for computing the evolution of cool white dwarfs employing detailed non-gray model atmospheres for pure H composition. We also explore the impact on the white dwarf cooling times of different assumptions for energy transfer in the atmosphere of cool white dwarfs. Detailed non-gray model atmospheres are computed taken into account non-ideal effects in the gas equation of state and chemical equilibrium, collision-induced absorption from molecules, and the Lyman alpha quasi-molecular opacity. Our results show that the use of detailed outer boundary conditions becomes relevant for effective temperatures lower than 5800 and 6100K for sequences with 0.60 and 0.90 M_sun, respectively. Detailed model atmospheres predict ages that are up to approx 10% shorter at log L/L_sun=-4 when compared with the ages derived using Eddington-like approximations at tau_Ross=2/3. We also analyze the effects of various assumptions and physical processes of relevance in the calculation of outer boundary conditions. In particular, we find that the Ly_alpha red wing absorption does not affect substantially the evolution of white dwarfs. White dwarf cooling timescales are sensitive to the surface boundary conditions for T_eff < 6000K. Interestingly enough, non-gray effects have little consequences on these cooling times at observable luminosities. In fact, collision-induced absorption processes, which significantly affect the spectra and colors of old white dwarfs with hydrogen-rich atmospheres, have not noticeable effects in their cooling rates, except throughout the Rosseland mean opacity.Comment: 6 pages, 9 figures, to be published in Astronomy and Astrophysic