Metabolism and toxicity of two new benzodiazepine-type antileishmanial agents in hepatocytes and macrophages

With increasing reports of resistance of Leishmania to antimonials (Thakur et al., 2004) and other traditional antileishmanial drugs, the need for the discovery of new antileishmanial agents is rising. In an attempt to find new antileishmanial agents, two new benzodiazepine (BNZ) analogues (7-chloro-4-(cyclohexylmethyl)-1-methyl-3,4-dihydro-1H-1,4-benodiazepine-2,5-dione (BNZ-1) and 4-(cyclohexylmethyl)-1-methyl-3,4-dihydro-1H-1,4-benzodiazepine-2,5-dione (BNZ-2)) were synthesised, and found to be effective against leishmaniasis in mice. This study investigates the metabolism of these two drugs together with the prototype BNZ, flurazepam (FZP), using rat hepatocytes, and investigates their interaction with glutathione in macrophages. Hepatocytes (>80% viability by Trypan Blue exclusion isolated by liver perfusion with collagenase) were prepared from male Sprague-Dawley rats (200-250 g). Drugs (100 μM) were incubated with 2 × 106 viable cells/ml in Krebs-Hepes buffer, pH 7.4, in rotating round bottomed flasks under an atmosphere of 95% O2/5% CO2, at 37 °C for 3 h, and timed samples taken for metabolite measurement. Samples were extracted twice with ethyl acetate at pH 10, the combined organic phases evaporated to dryness and stored at −20 °C until analysis. Metabolites were separated by HPLC using an ACE C18 column (50 mm × 3.0 mm i.d., 5 μm packing), and a solvent gradient consisting of 0.1% formic acid: acetonitrile (starting composition 95:5%, and composition after 25 min 65:35% for FZP and 70:30% for both BNZ 1 and 2). Flow rate was 0.5 ml/min, and detection was at 230 nm. Identification of the metabolites was by mass spectrometry with both positive and negative ion electronspray ionization. The effects of 24 h exposure to the compounds (100 μM) was investigated in the macrophage cell line J774.1 in terms of reduced glutathione content (GSH) and the activity of glutathione reductase (GR). There was no evidence of significant cytotoxicity with any of the compounds at the concentration used. FZP (m/z 388) was metabolised by dealkylation of the two N-1 ethyl substituents (m/z 360 and m/z 332), followed by hydroxylation on the BNZ ring. There was no evidence for either N- or O-glucuronidation of the resulting metabolites. BNZ-1 (m/z 321) was metabolised by N-demethylation (m/z 307) followed by hydroxylation (m/z 323), mono- and di-hydroxylation of the parent (m/z 337 and m/z 353) and by glucuronidation of the mono-hydroxylated metabolite (m/z 513). BNZ-2 (m/z 287) was transformed by N-demethylation (m/z 273) and hydroxylation of the parent (m/z 303), with the latter further metabolised by O-glucuronidation (m/z 479). In addition, the hydroxylated N-demethylated product (m/z 289) was also formed. Macrophages did not produce detectable metabolism of any of the drugs. All the drugs depleted macrophage GSH significantly (p < 0.05 by ANOVA followed by Dunnett's test) with BNZ-1 and BNZ-2 causing greater depletion than FZP (40.6 ± 4.0 and 45.8 ± 8.4, respectively, compared with 55.5 ± 4.9 nmol/mg protein with FZP, n = 3). Control macrophage GSH was 74.1 ± 6.6 nmol/mg protein. The depletion in GSH was not due to inhibition of GR: only FZP caused a significant decrease in macrophage GR activity (28.0 ± 5.9 compared with 42.1 ± 8.0 nmol/ml/min in control cells, p < 0.05 by ANOVA followed by Dunnett's test, n = 3). The marked depletion of macrophage GSH indicates a potential toxic interaction in mammalian cells. The new BNZ analogues were rapidly metabolised by hepatocytes, producing N-dealkylated and multiple hydroxylated phase I metabolites, followed by glucuronidation. This rapid metabolism may limit the therapeutic effect of BNZ 1 and 2 if their metabolites are inactive against leishmaniasis and suggest the need to optimise these lead structures further to obtain compounds with reduced rates and extent of metabolism

Solitons and Vertex Operators in Twisted Affine Toda Field Theories

Affine Toda field theories in two dimensions constitute families of integrable, relativistically invariant field theories in correspondence with the affine Kac-Moody algebras. The particles which are the quantum excitations of the fields display interesting patterns in their masses and coupling and which have recently been shown to extend to the classical soliton solutions arising when the couplings are imaginary. Here these results are extended from the untwisted to the twisted algebras. The new soliton solutions and their masses are found by a folding procedure which can be applied to the affine Kac-Moody algebras themselves to provide new insights into their structures. The relevant foldings are related to inner automorphisms of the associated finite dimensional Lie group which are calculated explicitly and related to what is known as the twisted Coxeter element. The fact that the twisted affine Kac-Moody algebras possess vertex operator constructions emerges naturally and is relevant to the soliton solutions.Comment: 27 pages (harvmac) + 3 figures (LaTex) at the end of the file, Swansea SWAT/93-94/1

Coronal Magnetic Field Evolution from 1996 to 2012: Continuous Non-potential Simulations

Coupled flux transport and magneto-frictional simulations are extended to simulate the continuous magnetic-field evolution in the global solar corona for over 15 years, from the start of Solar Cycle 23 in 1996. By simplifying the dynamics, our model follows the build-up and transport of electric currents and free magnetic energy in the corona, offering an insight into the magnetic structure and topology that extrapolation-based models cannot. To enable these extended simulations, we have implemented a more efficient numerical grid, and have carefully calibrated the surface flux-transport model to reproduce the observed large-scale photospheric radial magnetic field, using emerging active regions determined from observed line-of-sight magnetograms. This calibration is described in some detail. In agreement with previous authors, we find that the standard flux-transport model is insufficient to simultaneously reproduce the observed polar fields and butterfly diagram during Cycle 23, and that additional effects must be added. For the best-fit model, we use automated techniques to detect the latitude–time profile of flux ropes and their ejections over the full solar cycle. Overall, flux ropes are more prevalent outside of active latitudes but those at active latitudes are more frequently ejected. Future possibilities for space-weather prediction with this approach are briefly assessed

The LHCb Upgrade I

The LHCb upgrade represents a major change of the experiment. The detectors have been almost completely renewed to allow running at an instantaneous luminosity five times larger than that of the previous running periods. Readout of all detectors into an all-software trigger is central to the new design, facilitating the reconstruction of events at the maximum LHC interaction rate, and their selection in real time. The experiment's tracking system has been completely upgraded with a new pixel vertex detector, a silicon tracker upstream of the dipole magnet and three scintillating fibre tracking stations downstream of the magnet. The whole photon detection system of the RICH detectors has been renewed and the readout electronics of the calorimeter and muon systems have been fully overhauled. The first stage of the all-software trigger is implemented on a GPU farm. The output of the trigger provides a combination of totally reconstructed physics objects, such as tracks and vertices, ready for final analysis, and of entire events which need further offline reprocessing. This scheme required a complete revision of the computing model and rewriting of the experiment's software

The Origin, Early Evolution and Predictability of Solar Eruptions

Coronal mass ejections (CMEs) were discovered in the early 1970s when space-borne coronagraphs revealed that eruptions of plasma are ejected from the Sun. Today, it is known that the Sun produces eruptive flares, filament eruptions, coronal mass ejections and failed eruptions; all thought to be due to a release of energy stored in the coronal magnetic field during its drastic reconfiguration. This review discusses the observations and physical mechanisms behind this eruptive activity, with a view to making an assessment of the current capability of forecasting these events for space weather risk and impact mitigation. Whilst a wealth of observations exist, and detailed models have been developed, there still exists a need to draw these approaches together. In particular more realistic models are encouraged in order to asses the full range of complexity of the solar atmosphere and the criteria for which an eruption is formed. From the observational side, a more detailed understanding of the role of photospheric flows and reconnection is needed in order to identify the evolutionary path that ultimately means a magnetic structure will erupt

Measurement of the CP-Violating Asymmetry Amplitude sin2β\beta

We present results on time-dependent CP-violating asymmetries in neutral B decays to several CP eigenstates. The measurements use a data sample of about 88 million Y(4S) --> B Bbar decays collected between 1999 and 2002 with the BABAR detector at the PEP-II asymmetric-energy B Factory at SLAC. We study events in which one neutral B meson is fully reconstructed in a final state containing a charmonium meson and the other B meson is determined to be either a B0 or B0bar from its decay products. The amplitude of the CP-violating asymmetry, which in the Standard Model is proportional to sin2beta, is derived from the decay-time distributions in such events. We measure sin2beta = 0.741 +/- 0.067 (stat) +/- 0.033 (syst) and |lambda| = 0.948 +/- 0.051 (stat) +/- 0.017 (syst). The magnitude of lambda is consistent with unity, in agreement with the Standard Model expectation of no direct CP violation in these modes

Risk profiles and one-year outcomes of patients with newly diagnosed atrial fibrillation in India: Insights from the GARFIELD-AF Registry.

BACKGROUND: The Global Anticoagulant Registry in the FIELD-Atrial Fibrillation (GARFIELD-AF) is an ongoing prospective noninterventional registry, which is providing important information on the baseline characteristics, treatment patterns, and 1-year outcomes in patients with newly diagnosed non-valvular atrial fibrillation (NVAF). This report describes data from Indian patients recruited in this registry. METHODS AND RESULTS: A total of 52,014 patients with newly diagnosed AF were enrolled globally; of these, 1388 patients were recruited from 26 sites within India (2012-2016). In India, the mean age was 65.8 years at diagnosis of NVAF. Hypertension was the most prevalent risk factor for AF, present in 68.5% of patients from India and in 76.3% of patients globally (P < 0.001). Diabetes and coronary artery disease (CAD) were prevalent in 36.2% and 28.1% of patients as compared with global prevalence of 22.2% and 21.6%, respectively (P < 0.001 for both). Antiplatelet therapy was the most common antithrombotic treatment in India. With increasing stroke risk, however, patients were more likely to receive oral anticoagulant therapy [mainly vitamin K antagonist (VKA)], but average international normalized ratio (INR) was lower among Indian patients [median INR value 1.6 (interquartile range {IQR}: 1.3-2.3) versus 2.3 (IQR 1.8-2.8) (P < 0.001)]. Compared with other countries, patients from India had markedly higher rates of all-cause mortality [7.68 per 100 person-years (95% confidence interval 6.32-9.35) vs 4.34 (4.16-4.53), P < 0.0001], while rates of stroke/systemic embolism and major bleeding were lower after 1 year of follow-up. CONCLUSION: Compared to previously published registries from India, the GARFIELD-AF registry describes clinical profiles and outcomes in Indian patients with AF of a different etiology. The registry data show that compared to the rest of the world, Indian AF patients are younger in age and have more diabetes and CAD. Patients with a higher stroke risk are more likely to receive anticoagulation therapy with VKA but are underdosed compared with the global average in the GARFIELD-AF. CLINICAL TRIAL REGISTRATION-URL: http://www.clinicaltrials.gov. Unique identifier: NCT01090362

APOE ɛ4 exacerbates age-dependent deficits in cortical microstructure

The apolipoprotein E ɛ4 allele is the primary genetic risk factor for the sporadic type of Alzheimer’s disease. However, the mechanisms by which apolipoprotein E ɛ4 are associated with neurodegeneration are still poorly understood. We applied the Neurite Orientation Dispersion Model to characterize the effects of apolipoprotein ɛ4 and its interactions with age and education on cortical microstructure in cognitively normal individuals. Data from 1954 participants were included from the PREVENT-Dementia and ALFA (ALzheimer and FAmilies) studies (mean age = 57, 1197 non-carriers and 757 apolipoprotein E ɛ4 carriers). Structural MRI datasets were processed with FreeSurfer v7.2. The Microstructure Diffusion Toolbox was used to derive Orientation Dispersion Index maps from diffusion MRI datasets. Primary analyses were focused on (i) the main effects of apolipoprotein E ɛ4, and (ii) the interactions of apolipoprotein E ɛ4 with age and education on lobar and vertex-wise Orientation Dispersion Index and implemented using Permutation Analysis of Linear Models. There were apolipoprotein E ɛ4 × age interactions in the temporo-parietal and frontal lobes, indicating steeper age-dependent Orientation Dispersion Index changes in apolipoprotein E ɛ4 carriers. Steeper age-related Orientation Dispersion Index declines were observed among apolipoprotein E ɛ4 carriers with lower years of education. We demonstrated that apolipoprotein E ɛ4 worsened age-related Orientation Dispersion Index decreases in brain regions typically associated with atrophy patterns of Alzheimer’s disease. This finding also suggests that apolipoprotein E ɛ4 may hasten the onset age of dementia by accelerating age-dependent reductions in cortical Orientation Dispersion Index

Diagnosis and management of Silver-Russell syndrome: First international consensus statement

This Consensus Statement summarizes recommendations for clinical diagnosis, investigation and management of patients with Silver-Russell syndrome (SRS), an imprinting disorder that causes prenatal and postnatal growth retardation. Considerable overlap exists between the care of individuals born small for gestational age and those with SRS. However, many specific management issues exist and evidence from controlled trials remains limited. SRS is primarily a clinical diagnosis; however, molecular testing enables confirmation of the clinical diagnosis and defines the subtype. A 'normal' result from a molecular test does not exclude the diagnosis of SRS. The management of children with SRS requires an experienced, multidisciplinary approach. Specific issues include growth failure, severe feeding difficulties, gastrointestinal problems, hypoglycaemia, body asymmetry, scoliosis, motor and speech delay and psychosocial challenges. An early emphasis on adequate nutritional status is important, with awareness that rapid postnatal weight gain might lead to subsequent increased risk of metabolic disorders. The benefits of treating patients with SRS with growth hormone include improved body composition, motor development and appetite, reduced risk of hypoglycaemia and increased height. Clinicians should be aware of possible premature adrenarche, fairly early and rapid central puberty and insulin resistance. Treatment with gonadotropin-releasing hormone analogues can delay progression of central puberty and preserve adult height potential. Long-term follow up is essential to determine the natural history and optimal management in adulthood

Global, regional, and national life expectancy, all-cause mortality, and cause-specific mortality for 249 causes of death, 1980�2015: a systematic analysis for the Global Burden of Disease Study 2015

Background Improving survival and extending the longevity of life for all populations requires timely, robust evidence on local mortality levels and trends. The Global Burden of Disease 2015 Study (GBD 2015) provides a comprehensive assessment of all-cause and cause-specific mortality for 249 causes in 195 countries and territories from 1980 to 2015. These results informed an in-depth investigation of observed and expected mortality patterns based on sociodemographic measures. Methods We estimated all-cause mortality by age, sex, geography, and year using an improved analytical approach originally developed for GBD 2013 and GBD 2010. Improvements included refinements to the estimation of child and adult mortality and corresponding uncertainty, parameter selection for under-5 mortality synthesis by spatiotemporal Gaussian process regression, and sibling history data processing. We also expanded the database of vital registration, survey, and census data to 14�294 geography�year datapoints. For GBD 2015, eight causes, including Ebola virus disease, were added to the previous GBD cause list for mortality. We used six modelling approaches to assess cause-specific mortality, with the Cause of Death Ensemble Model (CODEm) generating estimates for most causes. We used a series of novel analyses to systematically quantify the drivers of trends in mortality across geographies. First, we assessed observed and expected levels and trends of cause-specific mortality as they relate to the Socio-demographic Index (SDI), a summary indicator derived from measures of income per capita, educational attainment, and fertility. Second, we examined factors affecting total mortality patterns through a series of counterfactual scenarios, testing the magnitude by which population growth, population age structures, and epidemiological changes contributed to shifts in mortality. Finally, we attributed changes in life expectancy to changes in cause of death. We documented each step of the GBD 2015 estimation processes, as well as data sources, in accordance with Guidelines for Accurate and Transparent Health Estimates Reporting (GATHER). Findings Globally, life expectancy from birth increased from 61·7 years (95 uncertainty interval 61·4�61·9) in 1980 to 71·8 years (71·5�72·2) in 2015. Several countries in sub-Saharan Africa had very large gains in life expectancy from 2005 to 2015, rebounding from an era of exceedingly high loss of life due to HIV/AIDS. At the same time, many geographies saw life expectancy stagnate or decline, particularly for men and in countries with rising mortality from war or interpersonal violence. From 2005 to 2015, male life expectancy in Syria dropped by 11·3 years (3·7�17·4), to 62·6 years (56·5�70·2). Total deaths increased by 4·1 (2·6�5·6) from 2005 to 2015, rising to 55·8 million (54·9 million to 56·6 million) in 2015, but age-standardised death rates fell by 17·0 (15·8�18·1) during this time, underscoring changes in population growth and shifts in global age structures. The result was similar for non-communicable diseases (NCDs), with total deaths from these causes increasing by 14·1 (12·6�16·0) to 39·8 million (39·2 million to 40·5 million) in 2015, whereas age-standardised rates decreased by 13·1 (11·9�14·3). Globally, this mortality pattern emerged for several NCDs, including several types of cancer, ischaemic heart disease, cirrhosis, and Alzheimer's disease and other dementias. By contrast, both total deaths and age-standardised death rates due to communicable, maternal, neonatal, and nutritional conditions significantly declined from 2005 to 2015, gains largely attributable to decreases in mortality rates due to HIV/AIDS (42·1, 39·1�44·6), malaria (43·1, 34·7�51·8), neonatal preterm birth complications (29·8, 24·8�34·9), and maternal disorders (29·1, 19·3�37·1). Progress was slower for several causes, such as lower respiratory infections and nutritional deficiencies, whereas deaths increased for others, including dengue and drug use disorders. Age-standardised death rates due to injuries significantly declined from 2005 to 2015, yet interpersonal violence and war claimed increasingly more lives in some regions, particularly in the Middle East. In 2015, rotaviral enteritis (rotavirus) was the leading cause of under-5 deaths due to diarrhoea (146�000 deaths, 118�000�183�000) and pneumococcal pneumonia was the leading cause of under-5 deaths due to lower respiratory infections (393�000 deaths, 228�000�532�000), although pathogen-specific mortality varied by region. Globally, the effects of population growth, ageing, and changes in age-standardised death rates substantially differed by cause. Our analyses on the expected associations between cause-specific mortality and SDI show the regular shifts in cause of death composition and population age structure with rising SDI. Country patterns of premature mortality (measured as years of life lost YLLs) and how they differ from the level expected on the basis of SDI alone revealed distinct but highly heterogeneous patterns by region and country or territory. Ischaemic heart disease, stroke, and diabetes were among the leading causes of YLLs in most regions, but in many cases, intraregional results sharply diverged for ratios of observed and expected YLLs based on SDI. Communicable, maternal, neonatal, and nutritional diseases caused the most YLLs throughout sub-Saharan Africa, with observed YLLs far exceeding expected YLLs for countries in which malaria or HIV/AIDS remained the leading causes of early death. Interpretation At the global scale, age-specific mortality has steadily improved over the past 35 years; this pattern of general progress continued in the past decade. Progress has been faster in most countries than expected on the basis of development measured by the SDI. Against this background of progress, some countries have seen falls in life expectancy, and age-standardised death rates for some causes are increasing. Despite progress in reducing age-standardised death rates, population growth and ageing mean that the number of deaths from most non-communicable causes are increasing in most countries, putting increased demands on health systems. Funding Bill & Melinda Gates Foundation. © 2016 The Author(s). Published by Elsevier Ltd. This is an Open Access article under the CC BY licens