Complete analysis of the B-cell response to a protein antigen, from in vivo germinal centre formation to 3-D modelling of affinity maturation

Somatic hypermutation of immunoglobulin variable region genes occurs within germinal centres (GCs) and is the process responsible for affinity maturation of antibodies during an immune response. Previous studies have focused almost exclusively on the immune response to haptens, which may be unrepresentative of epitopes on protein antigens. In this study, we have exploited a model system that uses transgenic B and CD4<sup>+</sup> T cells specific for hen egg lysozyme (HEL) and a chicken ovalbumin peptide, respectively, to investigate a tightly synchronized immune response to protein antigens of widely differing affinities, thus allowing us to track many facets of the development of an antibody response at the antigen-specific B cell level in an integrated system <i>in</i> <i>vivo</i>. Somatic hypermutation of immunoglobulin variable genes was analysed in clones of transgenic B cells proliferating in individual GCs in response to HEL or the cross-reactive low-affinity antigen, duck egg lysozyme (DEL). Molecular modelling of the antibody–antigen interface demonstrates that recurring mutations in the antigen-binding site, selected in GCs, enhance interactions of the antibody with DEL. The effects of these mutations on affinity maturation are demonstrated by a shift of transgenic serum antibodies towards higher affinity for DEL in DEL-cOVA immunized mice. The results show that B cells with high affinity antigen receptors can revise their specificity by somatic hypermutation and antigen selection in response to a low-affinity, cross-reactive antigen. These observations shed further light on the nature of the immune response to pathogens and autoimmunity and demonstrate the utility of this novel model for studies of the mechanisms of somatic hypermutation

Long term mortality after a single treatment course with X-rays in patients treated for ankylosing spondylitis.

Mortality up to 1 January 1983 has been studied in 14,106 patients with ankylosing spondylitis given a single course of X-ray treatment during 1935-54. For neoplasms other than leukaemia or colon cancer, mortality was 28% greater than that of members of the general population of England and Wales, and this increase is likely to have been a direct consequence of the treatment. The proportional increase reached a maximum of 71% between 10.0 and 12.4 years after irradiation and then declined. There was only a 7% increase in mortality from these tumours more than 25.0 years after irradiation and only for cancer of the oesophagus was the relative risk significantly raised in this period. Neither the magnitude of the relative risk, nor its temporal pattern following treatment, were greatly influenced by the age of the patient at first treatment. For leukaemia there was a threefold increase in mortality that is also likely to have been due to the radiotherapy. The relative risk was at its highest between 2.5 and 4.9 years after the treatment and then declined, but the increase did not disappear completely, and the risk was still nearly twice that of the general population more than 25.0 years after treatment. There was some evidence that the risks of acute myeloid, acute lymphatic, and chronic myeloid leukaemia were all increased, but no evidence of any increase in chronic lymphatic leukaemia. The relative risk appeared to be greatest for acute myeloid leukaemia. For colon cancer, which is associated with spondylitis through a common association with ulcerative colitis, mortality was increased by 30%. For non-neoplastic conditions there was a 51% increase in mortality that was likely to be associated with the disease itself rather than its treatment. The increase was apparent for a wide range of diseases and was not confined to diseases that have been associated clinically with ankylosing spondylitis

Paper trials: a qualitative study exploring the place of portfolios in making revalidation recommendations for Responsible Officers

A portfolio of supporting information (SI) reflecting a doctors entire medical practice is now a central aspect of UK appraisal for revalidation. Medical revalidation, introduced in 2012, is an assessment of a doctors competence and passing results in a five yearly license to practice medicine. It assesses of a doctors professional development, workplace performance and reflection and aims to provide assurance that doctors are up-to-date and fit to practice. The dominant assessment mechanism is a portfolio. The content of the revalidation portfolio has been increasingly prescribed and the assessment of the SI is a fundamental aspect of the appraisal process which ultimately allows Responsible Officers (ROs) to make recommendations on revalidation. ROs, themselves doctors, were the first to undergo UK revalidation. This qualitative study explored the perceptions of ROs and their appraisers about the use of this portfolio of evidence in a summative revalidation appraisal.28 purposefully sampled London ROs were interviewed following their revalidation appraisal and 17 of their appraisers participated in focus groups and interviews. Thematic analysis was used to identify commonalities and differences of experience.SI was mostly easy to provide but there were challenges in gathering certain aspects. ROs did not understand in what quantities they should supply SI or what it should look like. Appraisers were concerned about making robust judgements based on the evidence supplied. A lack of reflection from the process of collating SI and preparing for appraisal was noted and learning came more from the appraisal interview itself.More explicit guidance must be available to both appraisee and appraiser about what SI is required, how much, how it should be used and, how it will be assessed. The role of SI in professional learning and revalidation must be clarified and further empirical research is required to examine how best to use this evidence to make judgments as part of this type of appraisal

Salt Links Dominate Affinity of Antibody HyHEL-5 for Lysozyme through Enthalpic Contributions*

The binding of murine monoclonal antibody HyHEL-5 to lysozyme has been the subject of extensive crystallographic, computational, and experimental investigations. The complex of HyHEL-5 with hen egg lysozyme (HEL) features salt bridges between Fab heavy chain residue Glu50, and Arg45 and Arg68 of HEL. This interaction has been predicted to play a dominant role in the association on the basis of molecular electrostatics calculations. The association of aspartic acid and glutamine mutants at position 50H of the cloned HyHEL-5 Fab with HEL and bobwhite quail lysozyme (BQL), an avian variant bearing an Arg68 ? Lys substitution in the epitope, was characterized by isothermal titration calorimetry and sedimentation equilibrium. Affinities for HEL were reduced by 400-fold (E50HD) and 40,000-fold (E50HQ) (??G� estimated at 4.0 and 6.4 kcal mol?1, respectively). The same mutations reduce affinity for BQL by only 7- and 55-fold, respectively, indicating a reduced specificity for HEL. The loss of affinity upon mutation is in each case primarily due to an unfavorable change in the enthalpy of the interaction; the entropic contribution is virtually unchanged. An enthalpy-entropy compensation exists for each interaction; ?H� decreases, while ?S� increases with temperature. The ?Cp for each mutant interaction is less negative than the wild-type. Mutant-cycle analysis suggests the mutations present in the HyHEL-5 Fab mutants are linked to those present in the BQL with coupling energies between 3 and 4 kcal mol?1

Report of the Horse Mackerel Exchange and Workshop 2006

Following a recommendation from PGCCDBS, a workshop on age calibration of horse mackerel was carried out. The workshop was preceded by an exchange. The objectives were: to improve the quality of horse mackerel readings by international calibration. In particular, attempt to resolve the observed differences between countries. Estimate the accuracy and precision of the age readings before and after the intercalibration. Take into account differences between areas and methods. Training of new horse mackerel readers

Flexible access to conformationally-locked bicyclic morpholines

A preparatively accessible route to a series of conformationally-locked bicyclic morpholines has been developed. This flexible approach allows for diversification in order for a small array of lead-like scaffolds to be synthesised from readily available key building blocks

Vortical and Wave Modes in 3D Rotating Stratified Flows: Random Large Scale Forcing

Utilizing an eigenfunction decomposition, we study the growth and spectra of energy in the vortical and wave modes of a 3D rotating stratified fluid as a function of $\epsilon = f/N$. Working in regimes characterized by moderate Burger numbers, i.e. $Bu = 1/\epsilon^2 < 1$ or $Bu \ge 1$, our results indicate profound change in the character of vortical and wave mode interactions with respect to $Bu = 1$. As with the reference state of $\epsilon=1$, for $\epsilon < 1$ the wave mode energy saturates quite quickly and the ensuing forward cascade continues to act as an efficient means of dissipating ageostrophic energy. Further, these saturated spectra steepen as $\epsilon$ decreases: we see a shift from $k^{-1}$ to $k^{-5/3}$ scaling for $k_f < k < k_d$ (where $k_f$ and $k_d$ are the forcing and dissipation scales, respectively). On the other hand, when $\epsilon > 1$ the wave mode energy never saturates and comes to dominate the total energy in the system. In fact, in a sense the wave modes behave in an asymmetric manner about $\epsilon = 1$. With regard to the vortical modes, for $\epsilon \le 1$, the signatures of 3D quasigeostrophy are clearly evident. Specifically, we see a $k^{-3}$ scaling for $k_f < k < k_d$ and, in accord with an inverse transfer of energy, the vortical mode energy never saturates but rather increases for all $k < k_f$. In contrast, for $\epsilon > 1$ and increasing, the vortical modes contain a progressively smaller fraction of the total energy indicating that the 3D quasigeostrophic subsystem plays an energetically smaller role in the overall dynamics.Comment: 18 pages, 6 figs. (abbreviated abstract

Quality and impact of appraisal for revalidation: the perceptions of London’s responsible officers and their appraisers

BACKGROUND: To evaluate NHS England London region's approach to the revalidation appraisal of responsible officers in London, exploring perceptions of the quality and impact of the appraisal process. Revalidation is the process which aims to ensure doctors in the UK are up-to-date and fit to practice medicine thus improving the quality of patient care. Revalidation recommendations are largely premised on the documentation included in annual appraisals, which includes the professional development a doctor has undertaken and supporting information about their practice. METHODS: A pan-London qualitative study exploring the views of responsible officers and their appraisers about the revalidation appraisal process. The study aimed to gain an in-depth understanding of the experiences and perceptions of the participants. Responsible officers were purposefully sampled to represent the broadest range of designated bodies. Data analysis generated themes pertaining to quality and impact of appraisal for revalidation with the potential to feed into and shape the evolving system under investigation. RESULTS: The central importance of highly skilled appraisers was highlighted. Both groups reported educational opportunities embedded within the appraisal process. Independent appraisers, not matched by clinical speciality or place of work, were considered to take a more objective view of a responsible officer's practice by providing an 'outsider perspective'. However, covering the breadth of roles, in sufficient depth, was challenging. Participants reported a bias favouring the appraisal of the responsible officer role above others including clinical work. Appraisal and revalidation was perceived to have the potential to improve the healthcare standards and support both personal development and institutional quality improvement. CONCLUSIONS: Responsible officers play a central role in the revalidation process. Getting responsible officer appraisal right is central to supporting those individuals to in turn support doctors and healthcare organisations in continuous quality improvement. The complexity and importance of the role of responsible officer may make achieving an appraisal of all roles of such individuals problematic. This evaluation suggests responsible officer appraisal was perceived as educational and effective

'20 days protected learning' - students' experiences of an Overseas Nurses Programme - 4 years on: A retrospective survey

Background From September 2005 the Nursing and Midwifery Council (NMC) introduced new arrangements for the registration of non-EU overseas nurses which requires all applicants to undertake '20 days of protected learning' time in the UK and for some, a period of supervised practice. A survey was undertaken at Bournemouth University, which offers a '20 days protected learning only' programme, to elicit overseas nurses' demographic details, experiences in completing the programme and their 'final destinations' once registered. Methods An online survey was devised which contained a mixture of tick box and open ended questions which covered demographic details, views on the programme and final destinations This was uploaded to www.SurveyMonkey.com and sent out to nurses who had completed the Overseas Nurses Programme (ONP) with Bournemouth University (n=1050). Quantiative data were analysed using descriptive statistics and the qualitative data were coded and analysed using content analysis . Results There were 251 respondents (27.7% response rate). The typical 'profile' of a nurse who responded to the survey was female, aged 25-40 years and had been qualified for more than 5 years with a bachelors degree. The majority came from Australia on a 2 year working holiday visa and the key final destination in the UK, on registration with the NMC, was working for an agency. There were five key findings regarding experience of the programe. Of those surveyed 61.2% did not feel it necessary to undergo an ONP; 71.6% felt that they should be able to complete the programme on-line in their own country; 64.2% that the ONP should only contain information about delivery of healthcare in UK and Legal and professional (NMC) issues; 57% that European nurses should also undergo the same programme and sit an IELTS test; and 68.2% that the programme was too theory orientated; and should have links to practice (21%). Conclusions The NMC set the admissions criteria for entry to the register and Standards for an ONP. The findings of this survey raise issues regarding the percieved value and use of this approach for overseas nurses, and it may be helpful to take this into account when considering future policy