Regulation of Myc and its localization to Histone Locus Body in Drosophila

Myc is a transcriptional factor required for normal growth and development in vertebrates and invertebrates alike. Loss of function mutations in Myc can cause embryonic lethality in mammals and larval death in flies whereas an increase in its activity can lead to tumorigenesis. Therefore, proper regulation of Myc is very important to ensure normal development. Regulation of Myc occurs by several context specific mechanisms. One such mechanism is the negative feedback autoregulation of Myc and this mechanism is lost in all tumorigenic cell lines. Like its mammalian homolog, the Drosophila Myc (dMyc) undergoes autoregulation in the presence of an ectopic myc gene leading to a Myc null phenotype. Polycomb (Pc), a chromatin binding repressor is required for Myc autoregulation. Upon Pc knockdown, levels of Su(z)2, a Pc group related protein increase significantly, suggesting that Pc represses Su(z)2. We show here that ectopic Su(z)2 can interfere with Myc autorepression and restore endogenous Myc levels as well as rescue larval lethality caused due to Myc autorepression. Su(z)2 does not however, affect general repression by Myc suggesting that repression of myc locus occurs by a different mechanism. During this study we observed that Myc protein forms distinct puncta in certain tissues. Upon investigating we found that these Myc "spots" localize to sub-nuclear organelles known as Histone Locus Body (HLB). HLBs are histone pre-mRNA processing centers formed at histone gene locus. We show here that Myc localizes to the HLBs only during the S phase. Since hisones are transcribed only during S phase we hypothesize that Myc aids in histone transcription, a novel role for Myc

Clinical and Genetic Features of Choroideremia in Childhood.

PURPOSE: To review the functional and anatomic characteristics of choroideremia in the pediatric population, aiming to describe the earliest features of the disease and to identify biomarkers useful for monitoring disease progression. DESIGN: Retrospective case series. PARTICIPANTS: Children diagnosed with choroideremia at a single institution. METHODS: Patients were identified using an electronic patient record system. Case notes and retinal imaging (color fundus photography [CFP], spectral-domain [SD] optical coherence tomography [OCT], and fundus autofluorescence [FAF]) then were reviewed. The results of genetic testing also were recorded. MAIN OUTCOME MEASURES: Presenting symptoms, visual acuity, fundus changes (CFP, SD OCT, FAF), and CHM sequencing results. RESULTS: Twenty-nine patients were identified with a mean age at referral of 9 years (range, 3-16 years). CHM mutations were identified in 15 of 19 patients tested. Nyctalopia was the predominant symptom (66%). Five of 29 patients were asymptomatic at presentation. At the final follow-up visit (mean age, 16 years; range, 7-26 years), most maintained excellent visual acuity (mean, 0.98±0.13 decimalized Snellen acuity). The first sign of retinopathy was widespread pigment clumping at the level of the retinal pigment epithelium (RPE). This later evolved to chorioretinal atrophy, most marked in the mid-peripheral retina. Peripapillary atrophy also was an early feature and was progressive in nature. Three different zones of FAF change were visible. Persistence of the inner retinal layers, detected by SD OCT, was visible at presentation in 15 of 27 patients. Subfoveal choroidal thickness decreased with age, whereas central retinal thickness increased over a similar interval. Four patients in whom visual acuity decreased over the follow-up period recorded a reduction in central retinal thickness. CONCLUSIONS: Progressive structural changes occur at a time when central visual function is maintained. Pigmentary changes at the level of the RPE occur early in the disease course. Peripapillary chorioretinal atrophy, central retinal thickness, and subfoveal choroidal thickness are likely to be valuable in monitoring disease progression and should be considered as potential biomarkers in future therapeutic trials

Inhibition of Cytochrome P450 2E1 and Cytochrome P450 2A6 by essential oils: tarragon (Artemisia dracunculus) and basil (Ocimum basilicum)

Cytochrome P450 enzymes are involved in the metabolism of foreign substances that are present within living organisms. These P450s are classified as Phase I metabolizing enzymes that are found mainly in the liver, they belong to a superfamily of heme containing monoxygenases that are required for metabolism for a number of xenobiotics. Mechanistic approach by these enzymes involves carrying out the oxidation of carbon and nitrogen groups usually resulting in the addition of an alcohol. Cytochrome P450 enzymes have been involved in the bioactivation of inactive compounds to active electrophiles that can contribute to the production of reactive oxygen species, such as superoxide, free radicals, and peroxides. These ROS can contribute to oxidative stress, which leads to hepatic cell necrosis, lipid peroxidation, and DNA adduct formation. This study focuses on two specific Cytochrome P450s, CYP2E1 and CYP2A6. These enzymes have shown to have one of the highest rates of uncoupling among all P450s, this uncoupling leads to the production of ROS. This study focuses on finding potential inhibitors that decrease the chances of these uncoupling reactions from occurring. Human, rat, and rabbit liver microsomes were used for these studies. A series of essential oils were screened with these P450s to determine their inhibitory affects on the enzyme. From these studies, tarragon (Artemisia dracunculus) and basil (Ocimum basilicum) showed the highest inhibitory affects on the enzyme. The major constituent for both these oils was estragole, this was also confirmed via GCMS testing, because of this it was thought that it was a major reason to why the enzyme activity is being inhibited. With the initial screening data showing inhibition of CYP2E1 and CYP2A6, next goal was to determine how potent these oils were and what is the mode of inhibition, reversible or irreversible. The results of CYP2E1 and CYP2A6 with human, rat, and rabbit concluded that both essential oils and estragole inhibited the activity of the enzyme. Estragole was the most potent inhibitor of both enzymes in all 3 species. The KI value for estragole with CYP2E1 in humans was 22.4 µM, in rat 143 µM, and in rabbit 108 µM. The KI value for estragole with CYP2A6 in humans was 27.5 µM and in rabbit was 49.3 µM. Results show that the mode of inhibition shown by both these essential oils and estragole was non-competitive and a reversible type of interaction occurred with both Cytochrome P450 enzymes

ACOUSTIC SIGNAL PROPAGATION CHARACTERIZATION OF CONDUIT NETWORKS

Analysis of acoustic signal propagation in conduit networks has been an important area of research in acoustics. One major aspect of analyzing conduit networks as acoustic channels is that a propagating signal suffers frequency dependent attenuation due to thermo-viscous boundary layer effects and the presence of impedance mismatches such as side branches. The signal attenuation due to side branches is strongly influenced by their numbers and dimensions such as diameter and length. Newly developed applications for condition based monitoring of underground conduit networks involve measurement of acoustic signal attenuation through tests in the field. In many cases the exact installation layout of the field measurement location may not be accessible or actual installation may differ from the documented layout. The lack of exact knowledge of numbers and lengths of side branches, therefore, introduces uncertainty in the measurements of attenuation and contributes to the random variable error between measured results and those predicted from theoretical models. There are other random processes in and around conduit networks in the field that also affect the propagation of an acoustic signal. These random processes include but are not limited to the presence of strong temperature and humidity gradients within the conduits, blockages of variable sizes and types, effects of aging such as cracks, bends, sags and holes, ambient noise variations and presence of variable layer of water. It is reasonable to consider that the random processes contributing to the error in the measured attenuation are independent and arbitrarily distributed. The error, contributed by a large number of independent sources of arbitrary probability distributions, is best described by an approximately normal probability distribution in accordance with the central limit theorem. Using an analytical approach to model the attenuating effect of each of the random variable sources can be very complex and may be intractable. A tractable approach is to develop an empirical model of the attenuation that has a stochastic component of a finite mean and variance to account for the random variable error akin to addition of a normally distributed random variable shadowing component in the path loss models of radio frequency (RF) wireless communication channels. This approach forms the crux of the present study. To develop an empirical model, a large number of measurements in conduit networks were made in the field and in a laboratory test set up to measure the variability of attenuation with variation in four parameters. These parameters include distance of the receiver from the source, frequency, numbers and lengths of side branches. Variation in signal attenuation with distance at each transmitted frequency is predicted by using linear regression through the scatter plot of the measured data. Variations in signal attenuation due to change in frequency, number and lengths of side branches are measured in the field and laboratory tests by comparing the reference transmitted pressure with the received pressure at either the open end or at some distance away from the source along the conduit length. Residuals between measured and predicted sound pressure levels are computed and tested for normal probability distribution through a graphical method as well as a statistical goodness of fit test for quantifiable results. The findings indicate that an empirical model of signal attenuation, which includes a normally distributed random variable component to account for random variable errors in the attenuation measurements, gives a more accurate prediction of received acoustic signal strength in a conduit compared to existing theoretical models

Advanced diagnostic genetic testing in inherited retinal disease: experience from a single tertiary referral centre in the UK National Health Service

In 2013, as part of our genetic investigation of patients with inherited retinal disease, we utilized multigene panel testing of 105 genes known to cause retinal disease in our patient cohorts. This test was performed in a UK National Health Service (NHS) accredited laboratory. The results of all multigene panel tests requested between 1.4.13 and 31.8.14 were retrospectively reviewed. All patients had been previously seen at Moorfields Eye Hospital, London, UK and diagnosed with an inherited retinal dystrophy after clinical examination and detailed retinal imaging. The results were categorized into three groups: (i) Testing helped establish a certain molecular diagnosis in 45 out of 115 (39%). Variants in USH2A (n = 6) and RP1 (n = 4) were most common. (ii) Definitive conclusions could not be drawn from molecular testing alone in 13 out of 115 (11%) as either insufficient pathogenic variants were discovered or those identified were not consistent with the phenotype. (iii) Testing did not identify any pathogenic variants responsible for the phenotype in 57 out of 115 (50%). Multigene panel testing performed in an NHS setting has enabled a molecular diagnosis to be confidently made in 40% of cases. Novel variants accounted for 38% of all identified variants. Detailed retinal phenotyping helped the interpretation of specific variants. Additional care needs to be taken when assessing polymorphisms in genes that have been infrequently associated with disease, as historical techniques were not as rigorous as contemporary ones. Future iterations of sequencing are likely to offer higher sensitivity, testing a broader range of genes, more rapidly and at a reduced cost

Knowledge of pelvic floor problems: a study of third trimester, primiparous women

INTRODUCTION AND HYPOTHESIS: Pelvic floor problems in women (urinary incontinence, faecal incontinence, uterovaginal prolapse) are common, and have an adverse effect on quality of life. We hypothesized that there is low knowledge of these problems amongst primiparous women in their third trimester of pregnancy. METHODS: We conducted a cross-sectional study in antenatal clinics of three hospitals in London, UK, from 2011 to 2013. Primiparous women aged ≥18 years and in the third trimester of pregnancy answered questions on pelvic floor problems. Knowledge scores were calculated based on the proportion of questions answered correctly. RESULTS: A total of 249 women completed the question set. The average knowledge score across all domains was low at 45 %. Scores were lowest for the less common problems of faecal incontinence (35 %) and prolapse (36 %). The score for urinary incontinence was higher at 63 %, but low when questions explored more detailed levels of knowledge (41 %). Knowledge scores were positively associated with both education to tertiary level and the use of books as the information source on pregnancy and delivery. Only 35 % of women cited antenatal classes as a source. CONCLUSIONS: Knowledge of pelvic floor problems is low amongst third-trimester, primiparous women in this London-based population. Adequate knowledge of these problems is important for women to be able to make informed choices about their antenatal care and to seek help if problems arise. The data suggest scope for health-care professionals to raise these issues early during pregnancy, and to help women access accurate sources of information

The role of rewilding in mitigating hydrological extremes: State of the evidence

Landscape rewilding has the potential to help mitigate hydrological extremes by allowing natural processes to function. Our systematic review assessed the evidence base for rewilding-driven mitigation of high and low flows. The review uncovers a lack of research directly addressing rewilding, but highlights research in analogue contexts which can, with caution, indicate the nature of change. There is a lack of before-after studies that enable deeper examination of temporal trajectories and legacy effects, and a lack of research on the scrub and shrubland habitats common in rewilding projects. Over twice as much evidence is available for high flows compared to low flows, and fewer than one third of studies address high and low flows simultaneously, limiting our understanding of co-benefits and contrasting effects. Flow magnitude variables are better represented within the literature than flow timing variables, and there is greater emphasis on modeling for high flows, and on direct measurement for low flows. Most high flow studies report a mitigating effect, but with variability in the magnitude of effect, and some exceptions. The nature of change for low flows is more complex and suggests a higher potential for increased low flow risks associated with certain trajectories but is based on a very narrow evidence base. We recommend that future research aims to: capture effects on both high and low flow extremes for a given type of change; analyze both magnitude and timing characteristics of flow extremes; and examine temporal trajectories (before and after data) ideally using a full before-after-control-impact design. This article is categorized under: Human Water > Value of Water Science of Water > Hydrological Processes Science of Water > Water Extremes Water and Life > Conservation, Management, and Awareness

Large-scale analysis of antigenic diversity of T-cell epitopes in dengue virus

BACKGROUND: Antigenic diversity in dengue virus strains has been studied, but large-scale and detailed systematic analyses have not been reported. In this study, we report a bioinformatics method for analyzing viral antigenic diversity in the context of T-cell mediated immune responses. We applied this method to study the relationship between short-peptide antigenic diversity and protein sequence diversity of dengue virus. We also studied the effects of sequence determinants on viral antigenic diversity. Short peptides, principally 9-mers were studied because they represent the predominant length of binding cores of T-cell epitopes, which are important for formulation of vaccines. RESULTS: Our analysis showed that the number of unique protein sequences required to represent complete antigenic diversity of short peptides in dengue virus is significantly smaller than that required to represent complete protein sequence diversity. Short-peptide antigenic diversity shows an asymptotic relationship to the number of unique protein sequences, indicating that for large sequence sets (~200) the addition of new protein sequences has marginal effect to increasing antigenic diversity. A near-linear relationship was observed between the extent of antigenic diversity and the length of protein sequences, suggesting that, for the practical purpose of vaccine development, antigenic diversity of short peptides from dengue virus can be represented by short regions of sequences (~<100 aa) within viral antigens that are specific targets of immune responses (such as T-cell epitopes specific to particular human leukocyte antigen alleles). CONCLUSION: This study provides evidence that there are limited numbers of antigenic combinations in protein sequence variants of a viral species and that short regions of the viral protein are sufficient to capture antigenic diversity of T-cell epitopes. The approach described herein has direct application to the analysis of other viruses, in particular those that show high diversity and/or rapid evolution, such as influenza A virus and human immunodeficiency virus (HIV)

The Fundus Phenotype Associated With The p.Ala243Val BEST1 Mutation

PURPOSE: To describe a highly recognizable and reproducible retinal phenotype associated with a specific BEST1 mutation-p.Ala243Val. METHODS: Retrospective review of consecutive cases where genetic testing has identified p.Ala243Val BEST1 as the cause of disease. Electronic patient records were used to extract demographic, as well as functional and anatomical data. These data were compared with those observed with the most common BEST1 genotype, p.Arg218Cys. RESULTS: Eight individuals (six families) were identified with the p.Ala243Val BEST1 mutation and seven patients with the pathologic variant p.Arg218Cys. No patients with mutation of codon 243 knowingly had a family history of retinal disease, whereas all patients with the p.Arg218Cys variant did. The maculopathy was bilateral in all cases. The p.Ala243Val mutation was associated with a pattern dystrophy-type appearance, most visible with near-infrared reflectance and fundus autofluorescence imaging. This phenotype was never observed with any other genotype. This mutation was associated with an older median age of symptom onset (median = 42, interquartile range = 22) compared with those harboring the p.Arg218Cys mutation (median = 18, interquartile range = 12; Mann-Whitney U test; P 0.05). CONCLUSION: The mutation p.Ala243Val is associated with highly recognizable and reproducible pattern dystrophy-like phenotype. Patients develop symptoms at a later age and tend to have better preservation of electrooculogram amplitudes