The binding affinity of human IgG for its high affinity Fc receptor is determined by multiple amino acids in the CH2 domain and is modulated by the hinge region.

A family of chimeric immunoglobulins (Igs) bearing the murine variable region directed against the hapten dansyl linked to human IgG1, -2, -3, and -4 has been characterized with respect to binding to the human high affinity Fc gamma receptor, Fc gamma RI. Chimeric IgG1 and -3 have the highest affinity association (Ka = 10(9) M-1), IgG4 is 10-fold reduced from this level, and IgG2 displays no detectable binding. A series of genetic manipulations was undertaken in which domains from the strongly binding subclass IgG3 were exchanged with domains from the nonbinding subclass IgG2. The subclass of the CH2 domain was found to be critical for determining IgG receptor affinity. In addition, the hinge region was found to modulate the affinity of the IgG for Fc gamma RI, possibly by determining accessibility of Fc gamma RI to the binding site on Fc. A series of amino acid substitutions were engineered into the CH2 domain of IgG3 and IgG4 at sites considered potentially important to Fc receptor binding based on homology comparisons of binding and nonbinding IgG subclasses. Characterization of these mutants has revealed the importance for Fc gamma RI association of two regions of the genetic CH2 domain separated in primary structure by nearly 100 residues. The first of these is the hinge-link or lower hinge regions, in which two residues, Leu (234) and Leu(235) in IgG1 and -3, are critical to high affinity binding. Substitution at either of these sites reduces the IgG association constant by 10-100-fold. The second region that appears to contribute to receptor binding is in a hinge-proximal bend between two beta strands within the CH2 domain, specifically, Pro(331) in IgG1 and -3. As a result of beta sheet formation within this domain, this residue lies within 11 A of the hinge-link region. Substitution at this site reduces the Fc receptor association constant by 10-fold

Structural features of human immunoglobulin G that determine isotype-specific differences in complement activation.

Although very similar in sequence, the four subclasses of human immunoglobulin G (IgG) differ markedly in their ability to activate complement. Glu318-Lys320-Lys322 has been identified as a key binding motif for the first component of complement, C1q, and is present in all isotypes of Ig capable of activating complement. This motif, however, is present in all subclasses of human IgG, including those that show little (IgG2) or even no (IgG4) complement activity. Using point mutants of chimeric antibodies, we have identified specific residues responsible for the differing ability of the IgG subclasses to fix complement. In particular, we show that Ser at position 331 in gamma 4 is critical for determining the inability of that isotype to bind C1q and activate complement. Additionally, we provide further evidence that levels of C1q binding do not necessarily correlate with levels of complement activity, and that C1q binding alone is not sufficient for complement activation

The differential ability of human IgG1 and IgG4 to activate complement is determined by the COOH-terminal sequence of the CH2 domain.

Using domain switch chimeric antibodies, we confirm the important role of CH2 in complement activation. In addition, we demonstrate that the structures responsible for the differential ability of human IgG1 and IgG4 to activate complement are located at the COOH-terminal part (from residue 292 to 340) of the CH2 domain. The amino acids in CH2 that might be involved in complement interaction are discussed. While CH3 contributes to efficient complement activation, CH3 from IgG2 and CH3 IgG3 are equally effective

The effect of Am241 on UK plutonium recycle options in thorium-plutonium fuelled LWRs – Part I: PWRs

UK plutonium is expected to be managed using uranium-plutonium (U-Pu) mixed oxide (MOX) fuels in Light Water Reactors (LWRs). However, studies have shown that thorium-plutonium (Th-Pu) may be preferential. Research has mostly focussed on recycle of reactor grade Pu with limited minor actinide (MA) content. This study will determine if large quantities of americium (Am) in UK Pu may be restrictive to recycle schemes by determining the effect this has on reactivity feedback coefficients, fissile loading and incineration potential. Addition of Am is shown to result in predictable trends in reactivity feedback coefficients and spatial separation of Am and Pu is found to offer potential advantages over uniformly loaded fuel in terms of maximising fissile loading and incineration. Separation may also offer benefits in terms of targeting Am destruction, particularly if multiple recycle schemes are pursued, as this would maximise the fissile loading requirements while keeping reactivity feedback coefficients negative

The effect of Am241 on UK plutonium recycle options in thorium-plutonium fuelled LWRs – Part II: BWRs

UK plutonium is expected to be managed using uranium-plutonium (U-Pu) mixed oxide (MOX) fuels in Light Water Reactors (LWRs). However, studies have shown that thorium-plutonium (Th-Pu) may be preferential. Part I of this study considered the effect of americium (Am) in UK Pu in Pressurized Water Reactors (PWRs) and found that, while the reactivity response was sensitive to isotopic and spectral variations, trends were predictable. Part II focusses on separation of Am in Boiling Water Reactors (BWRs) and compares fuel performance to the uniformly distributed and spatially separated cases outlined in Part I. Comparable incineration rates are achievable but, while a single PWR assembly bears a greater mass of Am/Pu than a single BWR assembly, the full BWR core may be capable of operating with significantly greater fissile masses. Transmutation of Am241 to Am242 appears preferable to fast fission of Am241 as increased incineration occurs in lower void, bottom-of-assembly locations

Chemical generation and modification of peptides containing multiple dehydroalanines

Chemical formation of dehydroalanine has been widely used for the post-translational modification of protein and peptides, however methods to incorporate multiple dehydroalanine residues into a single peptide have not been defined. We report the use of methyl 2,5-dibromovalerate which can be used to cleanly carry out this transformation

Isotopic and spectral effects of Pu quality in Th-Pu fueled PWRs

UK plutonium (Pu) management is expected to focus on the use of uranium-plutonium (U-Pu) mixed oxide (MOX) fuel. However, research has shown that thorium-plutonium (Th-Pu) may be a viable alternative, offering favourable performance characteristics. A scoping study was carried out to determine the effect of isotopic composition and spectral hardening in standard and reduced moderation Pressurised Water Reactors (PWRs and RMPWRs). Lattice calculations were performed using WIMS to investigate safety parameters (Doppler Coefficient (DC), Moderator Temperature Coefficient (MTC), Void Coefficient (VC) – in this case Fully Voided Reactivity (FVR) – and Boron Worth (BW)), maximum theoretically achievable discharge burnup, Pu consumption and transuranic (TRU) composition of spent nuclear fuel (SNF) for the two reactor types. Standard grades of Pu were compared to a predicted UK Pu vector. MTC and FVR were found to be strongly influenced by the isotopic composition of the fuel. MTC was determined to be particularly sensitive to positive ‘peak’ contributions from fissile isotopes in the energy range 0.1–1 eV which diminish as the Pu content increases. The more extreme nature of the perturbation in FVR cases results in key differences in the contributions from fissile isotopes in the thermal energy range when compared with MTC, with no positive contributions from any isotope <500 eV. Where the requirement for MTC to remain negative was the limiting factor, a higher maximum fissile loading, discharge burnup and Pu consumption rate were possible in the PWR than the RMPWR, although the two reactors types typically produced similar levels of U233. However, for the majority of Pu grades the total minor actinide (MA) content in SNF was shown to be significantly lower in the RMPWR. Where FVR is the limiting factor, the maximum fissile loading and discharge burnup are similar in both reactor types, while increased Pu consumption rates were possible in the PWR. In this case, lower concentrations of U233 and MAs were found to be present in the PWR. These results are for a single pass of fuel through a reactor and, while the response of fissile isotopes at given energies to temperature perturbations will not vary significantly, the maximum achievable discharge burnup, Pu consumption rate and TRU build-up would be very different in a multi-recycle scenario.The first author is grateful for the financial support of the Engineering and Physical Sciences Research Council (EPSRC), Thor Energy, the Whitworth Society and the Institution of Engineering and Technology

Emerging pathogenic links between microbiota and the gut-lung axis

© 2016 Macmillan Publishers Limited, part of Springer Nature. All rights reserved. The microbiota is vital for the development of the immune system and homeostasis. Changes in microbial composition and function, termed dysbiosis, in the respiratory tract and the gut have recently been linked to alterations in immune responses and to disease development in the lungs. In this Opinion article, we review the microbial species that are usually found in healthy gastrointestinal and respiratory tracts, their dysbiosis in disease and interactions with the gut-lung axis. Although the gut-lung axis is only beginning to be understood, emerging evidence indicates that there is potential for manipulation of the gut microbiota in the treatment of lung diseases

Magnetic resonance imaging of placentome development in the pregnant Ewe

INTRODUCTION: Novel imaging measurements of placental development are difficult to validate due to the invasive nature of gold-standard procedures. Animal studies have been important in validation of magnetic resonance imaging (MRI) measurements in invasive preclinical studies, as they allow for controlled experiments and analysis of multiple time-points during pregnancy. This study characterises the longitudinal diffusion and perfusion properties of sheep placentomes using MRI, measurements that are required for future validation studies. METHODS: Pregnant ewes were anaesthetised for a MRI session on a 3T scanner. Placental MRI was used to classify placentomes morphologically into three types based on their shape and size at two gestational ages. To validate classification accuracy, placentome type derived from MRI data were compared with placentome categorisation results after delivery. Diffusion-Weighted MRI and T2-relaxometry were used to measure a broad range of biophysical properties of the placentomes. RESULTS: MRI morphological classification results showed consistent gestational age changes in placentome shape, as supported by post-delivery gold standard data. The mean apparent diffusion coefficient was significantly higher at 110 days gestation than at late gestation (~140 days; term, 150 days). Mean T2 was higher at mid gestation (152.2 ± 58.1 ms) compared to late gestation (127.8 ms ± 52.0). Significantly higher perfusion fraction was measured in late gestation placentomes that also had a significantly higher fractional anisotropy when compared to the earlier gestational age. DISCUSSION: We report baseline measurements of techniques common in placental MRI for the sheep placenta. These measurements are essential to support future validation measurements of placental MRI techniques

Sleep, circadian biology and skeletal muscle interactions: Implications for metabolic health

There currently exists a modern epidemic of sleep loss, triggered by the changing demands of our 21st century lifestyle that embrace 'round-the-clock' remote working hours, access to energy-dense food, prolonged periods of inactivity, and on-line social activities. Disturbances to sleep patterns impart widespread and adverse effects on numerous cells, tissues, and organs. Insufficient sleep causes circadian misalignment in humans, including perturbed peripheral clocks, leading to disrupted skeletal muscle and liver metabolism, and whole-body energy homeostasis. Fragmented or insufficient sleep also perturbs the hormonal milieu, shifting it towards a catabolic state, resulting in reduced rates of skeletal muscle protein synthesis. The interaction between disrupted sleep and skeletal muscle metabolic health is complex, with the mechanisms underpinning sleep-related disturbances on this tissue often multifaceted. Strategies to promote sufficient sleep duration combined with the appropriate timing of meals and physical activity to maintain circadian rhythmicity are important to mitigate the adverse effects of inadequate sleep on whole-body and skeletal muscle metabolic health. This review summarises the complex relationship between sleep, circadian biology, and skeletal muscle, and discusses the effectiveness of several strategies to mitigate the negative effects of disturbed sleep or circadian rhythms on skeletal muscle health