Studies on processing, particle formation, and immunogenicity of the HIV-1 gag gene product: a possible component of a HIV vaccine

Antigens in a particulate conformation were shown to be highly immunogenic in mammals. For this reason, the particle forming capacity of derivatives of the HIV-1 group specific core antigen p55 gag was assayed and compared dependent on various expression systems: recombinant bacteria, vaccinia- and baculoviruses were established encoding the entire core protein p55 either in its authentic sequence or lacking the myristylation consensus signal. Moreover, p55 gag was expressed in combination with the protease (p55-PR) or with the entire polymerase (p55-pol), respectively. Budding of 100-160 nm p55 core particles, resembling immature HIV-virions, was observed in the eucaryotic expression systems only. In comparison to the vaccinia virus driven expression of p55 in mammalian cells, considerably higher yields of particulate core antigen were obtained by infection of Spodoptera frugiperda (Sf9) insect cells with the recombinant Autographa californica nuclear polyhedrosis (AcMNPV) baculovirus. Mutation of the NH2-terminal myristylation signal sequence prevented budding of the immature core particles. Expression of the HIV p55-PR gene construct by recombinant baculovirus resulted in complete processing of the p55 gag precursor molecule in this system. The introduction of an artificial frameshift near the natural frameshift site resulted in constitutive expression of the viral protease and complete processing of p55, both in Escherichia coli and in vaccinia virus infected cells. Interestingly, significant processing of p55 resembling that of HIV infected H9 cells could also be achieved in the vaccinia system by fusing the entire pol gene to the gag gene. Moreover, processing was not found to be dependent on amino-terminal myristylation of the gag procursor molecule, which is in contrast to observations with type C and type D retrovirus. However, complete processing of p55 into p24, p17, p9 and p6 abolished particle formation. Purified immature HIV-virus like particles were highly immunogenic in rabbits, leading to a strong humoral immune response after immunization. Empty immature p55 gag particles represent a noninfectious and attractive candidate for a basic vaccine component

Effects of EDTA-Induced Hypocalcaemia and Stress on Plasma TNF-α, IL-1-ra, G-CSF, GM-CSF and S-100 in Dairy Cows

The pathophysiology of postparturient paresis is still not completely understood. Knowledge recently acquired in immunology, endocrinology and cell physiology has still to be integrated in order to elucidate the aetiopathogenesis of the disease. For that purpose, the effect of the EDTA infusion model on the plasma concentrations of selected cytokines and growth factors, and of a calcium binding protein was examined in dairy cows. Six 6- to 11-year-old Brown Swiss cows in mid lactation were infused with a 5% solution of Na2EDTA in one jugular vein over a period of 5 h. Blood samples were collected from the contralateral side daily two days before, and then hourly for five hours during the infusion, hourly for five hours after the end of the infusion, and once daily for 10 days thereafter. The plasma concentrations of cortisol, tumour necrosis factor-α, interleukin-1 receptor antagonist, granulocyte colony-stimulating factor, granulocyte and macrophage colony-stimulating factor, and the calcium binding protein S-100 were determined. Before the EDTA infusion, during the infusion and for two days thereafter, the mean plasma concentrations of cortisol were significantly higher than those from days 4 to 10 after the infusion. The plasma concentrations of tumour necrosis factor-α and interleukin-1 receptor antagonist followed a similar profile. At the end of EDTA infusion, low concentrations of granulocyte colony-stimulating factor were detected in one cow only. On days 3 and 4, the mean plasma concentrations of granulocyte colony-stimulating factor were significantly higher than the pre-infusion values, but this was followed by a significant decrease on post-infusion day 5. From day 4 to 7, the plasma concentrations of S-100 were significantly lower than the pre-infusion values. The importance of these findings in the pathophysiology of postparturient paresis remains to be establishe

Hydrodynamic dispersion within porous biofilms

Many microorganisms live within surface-associated consortia, termed biofilms, that can form intricate porous structures interspersed with a network of fluid channels. In such systems, transport phenomena, including flow and advection, regulate various aspects of cell behavior by controlling nutrient supply, evacuation of waste products, and permeation of antimicrobial agents. This study presents multiscale analysis of solute transport in these porous biofilms. We start our analysis with a channel-scale description of mass transport and use the method of volume averaging to derive a set of homogenized equations at the biofilm-scale in the case where the width of the channels is significantly smaller than the thickness of the biofilm. We show that solute transport may be described via two coupled partial differential equations or telegrapher's equations for the averaged concentrations. These models are particularly relevant for chemicals, such as some antimicrobial agents, that penetrate cell clusters very slowly. In most cases, especially for nutrients, solute penetration is faster, and transport can be described via an advection-dispersion equation. In this simpler case, the effective diffusion is characterized by a second-order tensor whose components depend on (1) the topology of the channels' network; (2) the solute's diffusion coefficients in the fluid and the cell clusters; (3) hydrodynamic dispersion effects; and (4) an additional dispersion term intrinsic to the two-phase configuration. Although solute transport in biofilms is commonly thought to be diffusion dominated, this analysis shows that hydrodynamic dispersion effects may significantly contribute to transport

Summer heat waves over western Europe 1880-2003, their relationship to large-scale forcings and predictability

We investigate the large-scale forcing and teleconnections between atmospheric circulation (sea level pressure, SLP), sea surface temperatures (SSTs), precipitation and heat wave events over western Europe using a new dataset of 54 daily maximum temperature time series. Forty four of these time series have been homogenised at the daily timescale to ensure that the presence of inhomogeneities has been minimised. The daily data have been used to create a seasonal index of the number of heat waves. Using canonical correlation analysis (CCA), heat waves over western Europe are shown to be related to anomalous high pressure over Scandinavia and central western Europe. Other forcing factors such as Atlantic SSTs and European precipitation, the later as a proxy for soil moisture, a known factor in strengthening land-atmosphere feedback processes, are also important. The strength of the relationship between summer SLP anomalies and heat waves is improved (from 35%) to account for around 46% of its variability when summer Atlantic and Mediterranean SSTs and summer European precipitation anomalies are included as predictors. This indicates that these predictors are not completely collinear rather that they each have some contribution to accounting for summer heat wave variability. However, the simplicity and scale of the statistical analysis masks this complex interaction between variables. There is some useful predictive skill of summer heat waves using multiple lagged predictors. A CCA using preceding winter North Atlantic SSTs and preceding January to May Mediterranean total precipitation results in significant hindcast (1972-2003) Spearman rank correlation skill scores up to 0.55 with an average skill score over the domain equal to 0.28±0.28. In agreement with previous studies focused on mean summer temperature, there appears to be some predictability of heat wave events on the decadal scale from the Atlantic Multidecadal Oscillation (AMO), although the long-term global mean temperature is also well related to western European heat waves. Combining these results with the observed positive trends in summer continental European SLP, North Atlantic SSTs and indications of a decline in European summer precipitation then possibly these long-term changes are also related to increased heat wave occurrence and it is important that the physical processes controlling these changes be more fully understoo

Passive-performance, analysis, and upgrades of a 1-ton seismic attenuation system

The 10m Prototype facility at the Albert-Einstein-Institute (AEI) in Hanover, Germany, employs three large seismic attenuation systems to reduce mechanical motion. The AEI Seismic-Attenuation-System (AEI-SAS) uses mechanical anti-springs in order to achieve resonance frequencies below 0.5Hz. This system provides passive isolation from ground motion by a factor of about 400 in the horizontal direction at 4Hz and in the vertical direction at 9Hz. The presented isolation performance is measured under vacuum conditions using a combination of commercial and custom-made inertial sensors. Detailed analysis of this performance led to the design and implementation of tuned dampers to mitigate the effect of the unavoidable higher order modes of the system. These dampers reduce RMS motion substantially in the frequency range between 10 and 100Hz in 6 degrees of freedom. The results presented here demonstrate that the AEI-SAS provides substantial passive isolation at all the fundamental mirror-suspension resonances

From Programme Theory to Logic Models for Multispecialty Community Providers: A Realist Evidence Synthesis

Background: The NHS policy of constructing multispecialty community providers (MCPs) rests on a complex set of assumptions about how health systems can replace hospital use with enhanced primary care for people with complex, chronic or multiple health problems, while contributing savings to health-care budgets. Objectives: To use policy-makers’ assumptions to elicit an initial programme theory (IPT) of how MCPs can achieve their outcomes and to compare this with published secondary evidence and revise the programme theory accordingly. Design: Realist synthesis with a three-stage method: (1) for policy documents, elicit the IPT underlying the MCP policy, (2) review and synthesise secondary evidence relevant to those assumptions and (3) compare the programme theory with the secondary evidence and, when necessary, reformulate the programme theory in a more evidence-based way. Data sources: Systematic searches and data extraction using (1) the Health Management Information Consortium (HMIC) database for policy statements and (2) topically appropriate databases, including MEDLINE, MEDLINE In-Process & Other Non-Indexed Citations, PsycINFO, the Cumulative Index to Nursing and Allied Health Literature (CINAHL) and Applied Social Sciences Index and Abstracts (ASSIA). A total of 1319 titles and abstracts were reviewed in two rounds and 116 were selected for full-text data extraction. We extracted data using a formal data extraction tool and synthesised them using a framework reflecting the main policy assumptions. Results: The IPT of MCPs contained 28 interconnected context–mechanism–outcome relationships. Few policy statements specified what contexts the policy mechanisms required. We found strong evidence supporting the IPT assumptions concerning organisational culture, interorganisational network management, multidisciplinary teams (MDTs), the uses and effects of health information technology (HIT) in MCP-like settings, planned referral networks, care planning for individual patients and the diversion of patients from inpatient to primary care. The evidence was weaker, or mixed (supporting some of the constituent assumptions but not others), concerning voluntary sector involvement, the effects of preventative care on hospital admissions and patient experience, planned referral networks and demand management systems. The evidence about the effects of referral reductions on costs was equivocal. We found no studies confirming that the development of preventative care would reduce demands on inpatient services. The IPT had overlooked certain mechanisms relevant to MCPs, mostly concerning MDTs and the uses of HITs. Limitations: The studies reviewed were limited to Organisation for Economic Co-operation and Development countries and, because of the large amount of published material, the period 2014–16, assuming that later studies, especially systematic reviews, already include important earlier findings. No empirical studies of MCPs yet existed. Conclusions: Multidisciplinary teams are a central mechanism by which MCPs (and equivalent networks and organisations) work, provided that the teams include the relevant professions (hence, organisations) and, for care planning, individual patients. Further primary research would be required to test elements of the revised logic model, in particular about (1) how MDTs and enhanced general practice compare and interact, or can be combined, in managing referral networks and (2) under what circumstances diverting patients from in-patient to primary care reduces NHS costs and improves the quality of patient experience

Characterisation of the mycobacterial NER system reveals novel functions of uvrD1 helicase

In this study, we investigated the role of the nucleotide excision repair (NER) pathway in mycobacterial DNA repair. Mycobacterium smegmatis lacking the NER excinuclease component uvrB, the helicase uvrD1 and a double knock-out lacking both proteins were constructed and their sensitivity to a series of DNA damaging agents wa analysed. As anticipated, the mycobacterial NER system was shown to be involved in the processing of bulky DNA adducts and inter-strand cross-links. In addition, it could be shown to exert a protective effect against oxidising and nitrosating agents. Interestingly, inactivation of uvrB and uvrD1 significantly increased marker integration frequencies in gene conversion assays. This implies that in mycobacteria, which lack the postreplicative mismatch repair system, NER, and particularly the UvrD1 helicase, is involved in the processing of a subset of recombination-associated mismatches

The importance of ship log data: reconstructing North Atlantic, European and Mediterranean sea level pressure fields back to 1750

Local to regional climate anomalies are to a large extent determined by the state of the atmospheric circulation. The knowledge of large-scale sea level pressure (SLP) variations in former times is therefore crucial when addressing past climate changes across Europe and the Mediterranean. However, currently available SLP reconstructions lack data from the ocean, particularly in the pre-1850 period. Here we present a new statistically-derived 5°×5° resolved gridded seasonal SLP dataset covering the eastern North Atlantic, Europe and the Mediterranean area (40°W-50°E; 20°N-70°N) back to 1750 using terrestrial instrumental pressure series and marine wind information from ship logbooks. For the period 1750-1850, the new SLP reconstruction provides a more accurate representation of the strength of the winter westerlies as well as the location and variability of the Azores High than currently available multiproxy pressure field reconstructions. These findings strongly support the potential of ship logbooks as an important source to determine past circulation variations especially for the pre-1850 period. This new dataset can be further used for dynamical studies relating large-scale atmospheric circulation to temperature and precipitation variability over the Mediterranean and Eurasia, for the comparison with outputs from GCMs as well as for detection and attribution studie