Characterization and Inhibition of a Class II Diterpene Cyclase from Mycobacterium tuberculosis: Implications for tuberculosis

Mycobacterium tuberculosis remains a widespread and devastating human pathogen, whose ability to infiltrate macrophage host cells from the human immune system is an active area of investigation. We have recently reported the discovery of a novel diterpene from M. tuberculosis, edaxadiene, whose ability to arrest phagosomal maturation in isolation presumably contributes to this critical process in M. tuberculosis infections. (Mann, F. M., Xu, M., Chen, X., Fulton, D. B., Russell, D. G., and Peters, R. J. (2009) J. Am. Chem. Soc., in press). Here, we present characterization of the class II diterpene cyclase that catalyzes the committed step in edaxadiene biosynthesis, i.e. the previously identified halimadienyl-diphosphate synthase (HPS; EC 5.5.1.16). Intriguingly, our kinetic analysis suggests a potential biochemical regulatory mechanism that triggers edaxadiene production upon phagosomal engulfment. Furthermore, we report characterization of potential HPS inhibitors: specifically, two related transition state analogs (15-aza-14,15-dihydrogeranylgeranyl diphosphate (7a) and 15-aza-14,15-dihydrogeranylgeranyl thiolodiphosphate (7b)) that exhibit very tight binding. Although arguably not suitable for clinical use, these nevertheless provide a basis for pharmaceutical design against this intriguing biosynthetic pathway. Finally, we provide evidence indicating that this pathway exists only in M. tuberculosis and is not functional in the closely related Mycobacterium bovis because of an inactivating frameshift in the HPS-encoding gene. Thus, we hypothesize that the inability to produce edaxadiene may be a contributing factor in the decreased infectivity and/or virulence of M. bovis relative to M. tuberculosis in humans

Interaction of Stress, Lead Burden, and Age on Cognition in Older Men: The VA Normative Aging Study

BACKGROUND. Low-level exposure to lead and to chronic stress may independently influence cognition. However, the modifying potential of psychosocial stress on the neurotoxicity of lead and their combined relationship to aging-associated decline have not been fully examined. OBJECTIVES. We examined the cross-sectional interaction between stress and lead exposure on Mini-Mental State Examination (MMSE) scores among 811 participants in the Normative Aging Study, a cohort of older U.S. men. METHODS. We used two self-reported measures of stress appraisal-a self-report of stress related to their most severe problem and the Perceived Stress Scale (PSS). Indices of lead exposure were blood lead and bone (tibia and patella) lead. RESULTS. Participants with higher self-reported stress had lower MMSE scores, which were adjusted for age, education, computer experience, English as a first language, smoking, and alcohol intake. In multivariable-adjusted tests for interaction, those with higher PSS scores had a 0.57-point lower (95% confidence interval, -0.90 to 0.24) MMSE score for a 2-fold increase in blood lead than did those with lower PSS scores. In addition, the combination of high PSS scores and high blood lead categories on one or both was associated with a 0.05-0.08 reduction on the MMSE for each year of age compared with those with low PSS score and blood lead level (p < 0.05). CONCLUSIONS. Psychological stress had an independent inverse association with cognition and also modified the relationship between lead exposure and cognitive performance among older men. Furthermore, high stress and lead together modified the association between age and cognition.National Institutes of Health (R01ES07821, R01HL080674, R01HL080674-02S1, R01ES013744, ES05257-06A1, P20MD000501, P42ES05947, ES03918-02); National Center for Research Resources General Clinical Research Center (M01RR02635); Leaves of Grass Foundation; United States Department of Veterans Affair

Notes on Decoherence at Absolute Zero

The problem of electron decoherence at low temperature is analyzed from the perspective of recent experiments on decoherence rate measurement and on related localization phenomena in low-dimensional systems. Importance of decoherence at zero temperature, perhaps induced by quantum fluctuations, is put in a broader context.Comment: 7 pages in PRB format, 1 figur

A Candidate Sub-Parsec Supermassive Binary Black Hole System

We identify SDSS J153636.22+044127.0, a QSO discovered in the Sloan Digital Sky Survey, as a promising candidate for a binary black hole system. This QSO has two broad-line emission systems separated by 3500 km/sec. The redder system at z=0.3889 also has a typical set of narrow forbidden lines. The bluer system (z=0.3727) shows only broad Balmer lines and UV Fe II emission, making it highly unusual in its lack of narrow lines. A third system, which includes only unresolved absorption lines, is seen at a redshift, z=0.3878, intermediate between the two emission-line systems. While the observational signatures of binary nuclear black holes remain unclear, J1536+0441 is unique among all QSOs known in having two broad-line regions, indicative of two separate black holes presently accreting gas. The interpretation of this as a bound binary system of two black holes having masses of 10^8.9 and 10^7.3 solar masses, yields a separation of ~ 0.1 parsec and an orbital period of ~100 years. The separation implies that the two black holes are orbiting within a single narrow-line region, consistent with the characteristics of the spectrum. This object was identified as an extreme outlier of a Karhunen-Loeve Transform of 17,500 z < 0.7 QSO spectra from the SDSS. The probability of the spectrum resulting from a chance superposition of two QSOs with similar redshifts is estimated at 2X10^-7, leading to the expectation of 0.003 such objects in the sample studied; however, even in this case, the spectrum of the lower redshift QSO remains highly unusual.Comment: 8 pages, 2 figures, Nature in pres

Indication of insensitivity of planetary weathering behavior and habitable zone to surface land fraction

It is likely that unambiguous habitable zone terrestrial planets of unknown water content will soon be discovered. Water content helps determine surface land fraction, which influences planetary weathering behavior. This is important because the silicate weathering feedback determines the width of the habitable zone in space and time. Here a low-order model of weathering and climate, useful for gaining qualitative understanding, is developed to examine climate evolution for planets of various land-ocean fractions. It is pointed out that, if seafloor weathering does not depend directly on surface temperature, there can be no weathering-climate feedback on a waterworld. This would dramatically narrow the habitable zone of a waterworld. Results from our model indicate that weathering behavior does not depend strongly on land fraction for partially ocean-covered planets. This is powerful because it suggests that previous habitable zone theory is robust to changes in land fraction, as long as there is some land. Finally, a mechanism is proposed for a waterworld to prevent complete water loss during a moist greenhouse through rapid weathering of exposed continents. This process is named a "waterworld self-arrest," and it implies that waterworlds can go through a moist greenhouse stage and end up as planets like Earth with partial ocean coverage. This work stresses the importance of surface and geologic effects, in addition to the usual incident stellar flux, for habitability.Comment: 15 pages, 6 figures, accepted at Ap

PML isoforms in response to arsenic: high-resolution analysis of PML body structure and degradation

Arsenic is a clinically effective treatment for acute promyelocytic leukaemia (APL) in which the promyelocytic leukaemia (PML) protein is fused to retinoic receptor alpha (RARα). PML-RARα is degraded by the proteasome by a SUMO-dependent, ubiquitin-mediated pathway in response to arsenic treatment, curing the disease. Six major PML isoforms are expressed as a result of alternative splicing, each of which encodes a unique C-terminal region. Using a system in which only a single EYFP-linked PML isoform is expressed, we demonstrate that PMLI, PMLII and PMLVI accumulate in the cytoplasm following arsenic treatment, whereas PMLIII, PMLIV and PMLV do not. 3D structured illumination was used to obtain super-resolution images of PML bodies, revealing spherical shells of PML along with associated SUMO. Arsenic treatment results in dramatic isoform-specific changes to PML body ultrastructure. After extended arsenic treatment most PML isoforms are degraded, leaving SUMO at the core of the nuclear bodies. A high-content imaging assay identifies PMLV as the isoform most readily degraded following arsenic treatment, and PMLIV as relatively resistant to degradation. Immunoprecipitation analysis demonstrates that all PML isoforms are modified by SUMO and ubiquitin after arsenic treatment, and by using siRNA, we demonstrate that arsenic-induced degradation of all PML isoforms is dependent on the ubiquitin E3 ligase RNF4. Intriguingly, depletion of RNF4 results in marked accumulation of PMLV, suggesting that this isoform is an optimal substrate for RNF4. Thus the variable C-terminal domain influences the rate and location of degradation of PML isoforms following arsenic treatment

IgG light chain-independent secretion of heavy chain dimers: consequence for therapeutic antibody production and design

Rodent monoclonal antibodies with specificity towards important biological targets are developed for therapeutic use by a process of humanisation. This process involves the creation of molecules, which retain the specificity of the rodent antibody but contain predominantly human coding sequence. Here we show that some humanised heavy chains can fold, form dimers and be secreted even in the absence of light chain. Quality control of recombinant antibody assembly in vivo is thought to rely upon folding of the heavy chain CH1 domain. This domain acts as a switch for secretion, only folding upon interaction with the light chain CL domain. We show that the secreted heavy-chain dimers contain folded CH1 domains and contribute to the heterogeneity of antibody species secreted during the expression of therapeutic antibodies. This subversion of the normal quality control process is dependent upon the heavy chain variable domain, is prevalent with engineered antibodies and can occur when only the Fab fragments are expressed. This discovery will impact on the efficient production of both humanised antibodies as well as the design of novel antibody formats

Secure and Internet-Less Connectivity to a Blockchain Network for Limited Connectivity Bank Users

Over the past few years, we have seen the emergence of a wide range of banking architectures, technologies, and applications made possible by the significant improvements in hardware, software, and networking technologies. Nowadays, innovative solutions are being developed by banks to leverage the benefits of blockchain, to improve their business agility and performance, and to make their business operations more efficient and secure. However, there are still cases where regular access to Internet is impossible or unreliable due to saturated networks or harsh environments, hence limiting the deployment of typical blockchain based solutions. In this context, an approach using a new connectivity technology is needed in order to increase mobile Internet services for any device to reach nearly 95% of the world population, instantly, simply by drawing on existing mobile phone networks, with no additional infrastructure development. We aim to give the user full bank access from their device, even if the device is not a smart one, using ordinary mobile phone networks. However, providing efficient and secure communications over lossy and low bandwidth networks remains a challenge. The main objective of this paper will be to design an end-to-end and low overhead secure solution for the communications between mobile devices and their corresponding remote application servers that using blockchain via ordinary mobile networks

Comparison of QG-Induced Dispersion with Standard Physics Effects

One of the predictions of quantum gravity phenomenology is that, in situations where Planck-scale physics and the notion of a quantum spacetime are relevant, field propagation will be described by a modified set of laws. Descriptions of the underlying mechanism differ from model to model, but a general feature is that electromagnetic waves will have non-trivial dispersion relations. A physical phenomenon that offers the possibility of experimentally testing these ideas in the foreseeable future is the propagation of high-energy gamma rays from GRB's at cosmological distances. With the observation of non-standard dispersion relations within experimental reach, it is thus important to find out whether there are competing effects that could either mask or be mistaken for this one. In this letter, we consider possible effects from standard physics, due to electromagnetic interactions, classical as well as quantum, and coupling to classical geometry. Our results indicate that, for currently observed gamma-ray energies and estimates of cosmological parameter values, those effects are much smaller than the quantum gravity one if the latter is first-order in the energy; some corrections are comparable in magnitude with the second-order quantum gravity ones, but they have a very different energy dependence.Comment: 8 pages; Version to be published in CQG as a letter; Includes some new comments and references, but no changes in the result