Deletion mutants in COP9/Signalosome subunits in fission yeast Schizosaccharomyces pombe display distinct phenotypes

The COP9/signalosome complex is highly conserved in evolution and possesses significant structural similarity to the 19S regulatory lid complex of the proteasome. It also shares limited similarity to the translation initiation factor eIF3. The signalosome interacts with multiple cullins in mammalian cells. In the fission yeast Schizosaccharomyces pombe, the Csn1 subunit is required for the removal of covalently attached Nedd8 from Pcu1, one of three S. pombe cullins. It remains unclear whether this activity is required for all the functions ascribed to the signalosome. We previously identified Csn1 and Csn2 as signalosome subunits in S. pombe. csn1 and csn2 null mutants are DNA damage sensitive and exhibit slow DNA replication. Two further putative subunits, Csn4 and Csn5, were identified from the S. pombe genome database. Herein, we characterize null mutations of csn4 and csn5 and demonstrate that both genes are required for removal of Nedd8 from the S. pombe cullin Pcu1 and that their protein products associate with Csn1 and Csn2. However, neither csn4 nor csn5 null mutants share the csn1 and csn2 mutant phenotypes. Our data suggest that the subunits of the signalosome cannot be considered as a distinct functional unit and imply that different subunits of the signalosome mediate distinct functions

Chromatin Structure in the Cellular Slime Mold Dictyostelium discoideum

The structure of Dictyostelium discoideum chromatin has been studied by the following techniques: electron microscopy, staphylococcal nuclease digestion, acrylamide gel electrophoresis, sucrose gradient centrifugation, and melting. The basic unit of chromatin is the nucleosome, which is a particle 98.6 angstrom in diameter. Approximately 50% of the chromatin is protected from nuclease digestion, but this decreases when protease activity is not inhibited. The nucleosome contains 187 base pairs of DNA, including a 137-base-pair core and a 50-base-pair linker. The monomer nucleosome has an s20,w value of 11.5 S on isokinetic sucrose gradients. When the chromatin is melted, four transitions are observed, at 54.5 degrees, 66.7 degrees, 74.9 degrees, and 79.7 degrees. The structure of Dictyostelium chromatin is very similar to that seen in higher eukaryotes

In situ correction of liquid meniscus in cell culture imaging system based on parallel Fourier ptychographic microscopy (96 Eyes)

We collaborated with Amgen and spent five years in designing and fabricating next generation multi-well plate imagers based on Fourier ptychographic microscopy (FPM). A 6-well imager (Emsight) and a low-cost parallel microscopic system (96 Eyes) based on parallel FPM were reported in our previous work. However, the effect of liquid meniscus on the image quality is much stronger than anticipated, introducing obvious wavevector misalignment and additional image aberration. To this end, an adaptive wavevector correction (AWC-FPM) algorithm and a pupil recovery improvement strategy are presented to solve these challenges in situ. In addition, dual-channel fluorescence excitation is added to obtain structural information for microbiologists. Experiments are demonstrated to verify their performances. The accuracy of angular resolution with our algorithm is within 0.003 rad. Our algorithms would make the FPM algorithm more robust and practical and can be extended to other FPM-based applications to overcome similar challenges

Evidence for potential underestimation of clinical folate deficiency in resource-limited countries using blood tests

Although a low serum folate concentration is a useful biomarker of pure folate deficiency, the presence of vitamin B12 deficiency or hemolysis or both in individuals with low folate status predictably raises serum folate levels. Therefore, in resource-limited settings where dietary folate deficiency can coexist with vitamin B12 deficiency or malaria or both, the serum folate concentration can range from normal to high, leading to serious underestimation of tissue folate status. This review traces the genesis of an inappropriate overreliance on the serum folate concentration to rule out folate deficiency in vulnerable populations of women and children. Of significance, without due consideration of a chronically inadequate dietary folate intake, authors of influential studies have likely wrongly judged these populations to have an adequate folate status. Through repetition, this error has led to a dangerous entry into the contemporary medical literature that folate deficiency is rare in women and children. As a consequence, many millions of under-resourced women and children with mild to moderate tissue folate deficiency may have been deprived of folate replacement. This review uses historical documents to challenge earlier conclusions and re-emphasizes the need for contextual integration of clinical information in resource-limited settings

Measuring the impact of the Capital Card®, a novel form of contingency management, on substance misuse treatment outcomes:A retrospective evaluation

Background The Capital Card, developed by WDP, is a digital innovation which acts as a form of contingency management, and aims to significantly improve service user outcomes. WDP is a substance misuse treatment provider commissioned by local authorities across the UK to support service users and their families affected by addiction. The Capital Card, much like commercial loyalty cards, uses a simple earn-spend points system which incentivises and rewards service users for engaging with services e.g. by attending key work sessions, Blood Borne Virus appointments or group-work sessions. The Spend activities available to service users are designed to improve overall wellbeing and build social and recovery capital, and include activities such as educational classes, fitness classes, driving lessons, and cinema tickets. Methods and findings We compared successful completion rates of 1,545 service users accessing one of WDP’s London based community services over a two-year period; before and after the Capital Card was introduced. Client demographics (age, sex and primary substance) were controlled for during the analysis. Once client demographics were controlled for, analysis showed that clients with a Capital Card were 1.5 times more likely to successfully complete treatment than those who had not had the Capital Card (OR = 1.507, 95% CI = 1.194 to 1.902). Conclusions The results of this initial evaluation are of particular interest to commissioners and policy makers as it indicates that the Capital Card can be used effectively as a form of contingency management to enhance recovery outcomes for service users engaging in community-based substance misuse services

Functional programming framework for GRworkbench

The software tool GRworkbench is an ongoing project in visual, numerical General Relativity at The Australian National University. Recently, the numerical differential geometric engine of GRworkbench has been rewritten using functional programming techniques. By allowing functions to be directly represented as program variables in C++ code, the functional framework enables the mathematical formalism of Differential Geometry to be more closely reflected in GRworkbench . The powerful technique of `automatic differentiation' has replaced numerical differentiation of the metric components, resulting in more accurate derivatives and an order-of-magnitude performance increase for operations relying on differentiation

Deuterium Abundance in the Most Metal-Poor Damped Lyman alpha System: Converging on Omega_baryons

The most metal-poor DLA known to date, at z = 2.61843 in the spectrum of the QSO Q0913+072, with an oxygen abundance only about 1/250 of the solar value, shows six well resolved D I Lyman series transitions in high quality echelle spectra recently obtained with the ESO VLT. We deduce a value of the deuterium abundance log (D/H) = -4.56+/-0.04 which is in good agreement with four out of the six most reliable previous determinations of this ratio in QSO absorbers. We find plausible reasons why in the other two cases the 1 sigma errors may have been underestimated by about a factor of two. The addition of this latest data point does not change significantly the mean value of the primordial abundance of deuterium, suggesting that we are now converging to a reliable measure of this quantity. We conclude that = -4.55+/-0.03 and Omega_b h^2 (BBN) = 0.0213+/-0.0010 (68% confidence limits). Including the latter as a prior in the analysis of the five year data of WMAP leads to a revised best-fitting value of the power-law index of primordial fluctuations n_s = 0.956+/-0.013 (1 sigma) and n_s < 0.990 with 99% confidence. Considering together the constraints provided by WMAP 5, (D/H)_p, baryon oscillations in the galaxy distribution, and distances to Type Ia supernovae, we arrive at the current best estimates Omega_b h^2 = 0.0224+/-0.0005 and n_s = 0.959+/-0.013.Comment: 13 pages, 8 Figures. Revised version following referee's comments. Accepted for publication in Monthly Notices of the Royal Astronomical Society. A few typos correcte

Reconstructing ice-sheet accumulation rates at ridge B, East Antarctica

Understanding how ice sheets responded to past climate change is fundamental to forecasting how they will respond in the future. Numerical models calculating the evolution of ice sheets depend upon accumulation data, which are principally available from ice cores. Here, we calculate past rates of ice accumulation using internal layering. The englacial structure of the East Antarctic ice divide at ridge B is extracted from airborne ice-penetrating radar. The isochronous surfaces are dated at their intersection with the Vostok ice-core site, where the depth–age relationship is known. The dated isochrons are used as input to a one-dimensional ice-flow model to investigate the spatial accumulation distribution. The calculations show that ice-accumulation rates generally increase from Vostok lake towards ridge B. The western flank of the ice divide experiences markedly more accumulation than in the east. Further, ice accumulation increases northwards along the ice divide. The results also show the variability of accumulation in time and space around the ridge B ice divide over the last 124 000 years

Individuals with low back pain: how do they view physical activity?

Background. Recent guidelines for those with acute low back pain have advocated early resumption of normal activity and increased physical activity. Little is known about the relationship between low back pain and physical activity, and on the impact of that relationship on the promotion of increased levels of physical activity within a general practice population. Objectives. We aimed to explore associations between factors that influence changes in physical activity and the way individuals perceive and behave with their low back pain, and the impact of those perceptions and behaviour on physical activity. Methods. Twenty-seven informants were chosen using a purposive sample from a larger group of individuals who, because of their low back trouble, had been referred by their GPs to a community-based, single-blind, randomized controlled trial (RCT) at the University of York, which is evaluating the effectiveness and cost-effectiveness of a progressive exercise programme. Fifty-four interviews were conducted with this subgroup of the RCT; four informants were interviewed once, 19 twice and four of them three times. Interviews were transcribed and analysed using manual and computer-aided approaches. Results. Physical activity was perceived as (i) activities of daily living, (ii) activities causing breathlessness that they went out of the way to do and (iii) more competitive-type activity. The avoidance of physical activity and fear of pain returning were the two main factors directly associated with informants' backs and changes in physical activity. These two factors hindered increases in physical activity, even though the majority of informants believed strongly that being physically active helped ease their low back pain. Conclusions. When advocating that individuals with acute low back pain return to or increase physical activity, it is important that clinicians identify avoidance of physical activity and/or fear of pain at the earliest stage in order to tailor advice and reassurance appropriately. If avoidance of activity and fear of pain is identified and clinicians want to encourage patients to take up and sustain increased physical activity, they should explore issues of fear of pain, and avoidance of and confidence to do physical activities, in addition to other factors influencing physical activity