Adenovirus serotype 5 L4-22K and L4-33K proteins have distinct functions in regulating late gene expression

Adenoviruses express up to 20 distinct mRNAs from five major late transcription unit (MLTU) regions, L1 to L5, by differential splicing and polyadenylation of the primary transcript. MLTU expression is regulated at transcriptional and posttranscriptional levels. The L4-33K protein acts as a splicing factor to upregulate several MLTU splice acceptor sites as the late phase progresses. The L4 region also expresses a 22K protein whose sequence is related to the sequence of L4-33K. L4-22K is shown here also to have an important role in regulating the pattern of MLTU gene expression. An adenovirus genome containing a stop codon in the L4-22K open reading frame expressed low levels of both structural and nonstructural late proteins compared to the wild-type (wt) adenovirus genome; a decrease in intermediate proteins, IVa2 and IX, was also observed. However, early protein synthesis and replication were unaffected by the absence of L4-22K. Intermediate and late protein expression was restored to wt levels by L4-22K expressed in trans but not by L4-33K. Increased MLTU promoter activity, resulting from stabilization of the transcriptional activator IVa2 by L4-22K, made a small contribution to this restoration of late gene expression. However, the principal effect of L4-22K was on the processing of MLTU RNA into specific cytoplasmic mRNA. L4-22K selectively increased expression of penton mRNA and protein, whereas splicing to create penton mRNA is known not to be increased by L4-33K. These results indicate that L4-22K plays a key role in the early-late switch in MLTU expression, additional to and distinct from the role of L4-33K

Discriminating between chronic fatigue syndrome and depression: a cognitive analysis

Background: chronic fatigue syndrome (CFS) and depression share a number of common symptoms and the majority of CFS patients meet lifetime criteria for depression. While cognitive factors seem key to the maintenance of CFS and depression, little is known about how the cognitive characteristics differ in the two conditions.Methods: fifty-three CFS patients were compared with 20 depressed patients and 38 healthy controls on perceptions of their health, illness attributions, self-esteem, cognitive distortions of general and somatic events, symptoms of distress and coping. A 6 month follow-up was also conducted to determine the stability of these factors and to investigate whether CFS-related cognitions predict ongoing disability and fatigue in this disorder.Results: between-group analyses confirmed that the depressed group was distinguished by low self-esteem, the propensity to make cognitive distortions across all situations, and to attribute their illness to psychological factors. In contrast, the CFS patients were characterized by low ratings of their current health status, a strong illness identity, external attributions for their illness, and distortions in thinking that were specific to somatic experiences. They were also more likely than depressed patients to cope with their illness by limiting stress and activity levels. These CFS-related cognitions and behaviours were associated with disability and fatigue 6 months later.Conclusions: CFS and depression can be distinguished by unique cognitive styles characteristic of each condition. The documented cognitive profile of the CFS patients provides support for the current cognitive behavioural models of the illness

Generation of cell lines to complement Adenovirus vectors using recombination-mediated cassette exchange

Background Adenovirus serotype 5 (Ad5) has many favourable characteristics for development as a gene therapy vector. However, the utility of current Ad5 vectors is limited by transient transgene expression, toxicity and immunogenicity. The most promising form of vector is the high capacity type, which is deleted for all viral genes. However, these vectors can only be produced to relatively low titres and with the aid of helper virus. Therefore a continuing challenge is the generation of more effective Ad5 vectors that can still be grown to high titres. Our approach is to generate complementing cell lines to support the growth of Ad5 vectors with novel late gene deficiencies. Results We have used LoxP/Cre recombination mediated cassette exchange (RMCE) to generate cell lines expressing Ad5 proteins encoded by the L4 region of the genome, the products of which play a pivotal role in the expression of Ad5 structural proteins. A panel of LoxP parent 293 cell lines was generated, each containing a GFP expression cassette under the control of a tetracycline-regulated promoter inserted at a random genome location; the cassette also contained a LoxP site between the promoter and GFP sequence. Clones displayed a variety of patterns of regulation, stability and level of GFP expression. Clone A1 was identified as a suitable parent for creation of inducible cell lines because of the tight inducibility and stability of its GFP expression. Using LoxP-targeted, Cre recombinase-mediated insertion of an L4 cassette to displace GFP from the regulated promoter in this parent clone, cell line A1-L4 was generated. This cell line expressed L4 100K, 22K and 33K proteins at levels sufficient to complement L4-33K mutant and L4-deleted viruses. Conclusions RMCE provides a method for rapid generation of Ad5 complementing cell lines from a pre-selected parental cell line, chosen for its desirable transgene expression characteristics. Parent cell lines can be selected for high or low gene expression, and for tight regulation, allowing viral protein expression to mirror that found during infection. Cell lines derived from a single parent will allow the growth of different vectors to be assessed without the complication of varying complementing protein expression

Saturated gain spectrum of VECSELs determined by transient measurement of lasing onset

We describe time-resolved measurements of the evolution of the spectrum of radiation emitted by an optically-pumped continuous-wave InGaAs-GaAs quantum well laser, recorded as lasing builds up from noise to steady state. We extract a fitting parameter corresponding to the gain dispersion of the parabolic spectrum equal to ?79 ± 30 fs2 and ?36 ± 6 fs2 for a resonant and anti-resonant structure, respectively. Furthermore the recorded evolution of the spectrum allows for the calculation of an effective FWHM gain bandwidth for each structure, of 11 nm and 18 nm, respectively

Syndemics and the PrEP Cascade: Results from a Sample of Young Latino Men Who Have Sex with Men.

Young Latino men who have sex with men (MSM) are a highly vulnerable population for HIV infection. Pre-exposure prophylaxis (PrEP) is a novel biomedical HIV prevention tool that may aid in reducing the disparity in HIV incidence among Latino MSM. However, PrEP use is disproportionally low among Latino MSM and, therefore, identifying barriers along the PrEP continuum of care (the "PrEP cascade") would provide insight into how to best deploy PrEP interventions. Syndemics theory is a prominent framework employed in HIV prevention; however, to date, no known studies have applied this theory to PrEP. Thus, the aim of the current study was to explore the association between syndemics and the PrEP cascade, including the degree to which psychosocial and structural syndemic constructs are related to the PrEP cascade. Participants were 151 young Latino MSM (M age = 24 years; SD = 3) residing in San Diego, California, who completed a battery of online self-report measures. Results indicated high levels of syndemic indicators and varying levels of engagement across the PrEP cascade. As syndemic indicators increased, the odds of engagement across the PrEP cascade were significantly lowered. Psychosocial and structural syndemic factors accounted for unique variance in the PrEP cascade. Results highlight the need for combination interventions that address both psychosocial and structural barriers to PrEP use and persistence among young Latino MSM

A phenomenological description of BslA assemblies across multiple length scales

Intrinsically interfacially active proteins have garnered considerable interest recently owing to their potential use in a range of materials applications. Notably, the fungal hydrophobins are known to form robust and well-organized surface layers with high mechanical strength. Recently, it was shown that the bacterial biofilm protein BslA also forms highly elastic surface layers at interfaces. Here we describe several self-assembled structures formed by BslA, both at interfaces and in bulk solution, over a range of length scales spanning from nanometres to millimetres. First, we observe transiently stable and highly elongated air bubbles formed in agitated BslA samples. We study their behaviour in a range of solution conditions and hypothesize that their dissipation is a consequence of the slow adsorption kinetics of BslA to an air–water interface. Second, we describe elongated tubules formed by BslA interfacial films when shear stresses are applied in both a Langmuir trough and a rheometer. These structures bear a striking resemblance, although much larger in scale, to the elongated air bubbles formed during agitation. Taken together, this knowledge will better inform the conditions and applications of how BslA can be used in the stabilization of multi-phase materials. This article is part of the themed issue ‘Soft interfacial materials: from fundamentals to formulation’

Discovery of 28 pulsars using new techniques for sorting pulsar candidates

Modern pulsar surveys produce many millions of candidate pulsars, far more than can be individually inspected. Traditional methods for filtering these candidates, based upon the signal-to-noise ratio of the detection, cannot easily distinguish between interference signals and pulsars. We have developed a new method of scoring candidates using a series of heuristics which test for pulsar-like properties of the signal. This significantly increases the sensitivity to weak pulsars and pulsars with periods close to interference signals. By applying this and other techniques for ranking candidates from a previous processing of the Parkes Multi-beam Pulsar Survey, 28 previously unknown pulsars have been discovered. These include an eccentric binary system and a young pulsar which is spatially coincident with a known supernova remnant.Comment: To be published in Monthly Notices of the Royal Astronomical Society. 11 pages, 9 figure

Soil organic carbon pool changes following land-use conversions

Includes bibliographical references (pages 1131-1132).Carbon (C) can be sequestered in the mineral soil after the conversion of intensively cropped agricultural fields to more extensive land uses such as afforested and natural succession ecosystems. Three land-use treatments from the long-term ecological research site at Kellogg biological station in Michigan were compared with a nearby deciduous forest. Treatments included a conventionally tilled cropland, a former cropland afforested with poplar for 10 years and an old field (10 years) succession. We used soil aggregate and soil organic matter fractionation techniques to isolate C pools that (1) have a high potential for C storage and (2) accumulate C at a fast rate during afforestation or succession. These fractions could serve as sensitive indicators for the total change in C content due to land-use changes. At the mineral soil surface (0–7 cm), afforesting significantly increased soil aggregation to levels similar to native forest. However, surface soil (0–7 cm) C did not follow this trend: soil C of the native forest site (22.9 t C ha-1) was still significantly greater than the afforested (12.6 t C ha-1) and succession (15.4 t C ha-1) treatments. However, when the 0–50 cm soil layer was considered, no differences in total soil C were observed between the cropland and the poplar afforested system, while the successional system increased total soil C (0–50 cm) at a rate of 0.786 t C ha-1 yr-1. Afforested soils sequestered C mainly in the fine intra-aggregate particulate organic matter (POM) (53–250 μm), whereas the successional soils sequestered C preferentially in the mineral-associated organic matter and fine intra-aggregate POM C pools

Feminism, Abortion and Disability: irreconcilable differences?

There has been considerable discussion of the political allegiance between the feminist and disability movements, but the question of abortion remains a thorny one. Disability rights advocates have been keen to demonstrate that it is possible to believe in a woman's right to sovereignty over the body and, yet, be opposed to the selective abortion of an impaired foetus – describing the latter as a form of 'weak' eugenics. The aim of this paper is to show that whilst there may be some points of agreement between the feminist and disability movements on the question of abortion, there exist fundamental and irreconcilable differences