Intracellular Antibodies for Drug Discovery and as Drugs of the Future.

The application of antibodies in cells was first shown in the early 1990s, and subsequently, the field of intracellular antibodies has expanded to encompass antibody fragments and their use in target validation and as engineered molecules that can be fused to moieties (referred to as warheads) to replace the Fc effector region of a whole immunoglobulin to elicit intracellular responses, such as cell death pathways or protein degradation. These various forms of intracellular antibodies have largely been used as research tools to investigate function within cells by perturbing protein activity. New applications of such molecules are on the horizon, namely their use as drugs per se and as templates for small-molecule drug discovery. The former is a potential new pharmacology that could harness the power and flexibility of molecular biology to generate new classes of drugs (herein referred to as macrodrugs when used in the context of disease control). Delivery of engineered intracellular antibodies, and other antigen-binding macromolecules formats, into cells to produce a therapeutic effect could be applied to any therapeutic area where regulation, degradation or other kinds of manipulation of target proteins can produce a therapeutic effect. Further, employing single-domain antibody fragments as competitors in small-molecule screening has been shown to enable identification of drug hits from diverse chemical libraries. Compounds selected in this way can mimic the effects of the intracellular antibodies that have been used for target validation. The capability of intracellular antibodies to discriminate between closely related proteins lends a new dimension to drug screening and drug development

Occupational exposure to blood in medical students

Objective. To determine the extent of occupational exposure to blood in medical students, details of the circumstances surrounding the incidents and the subsequent experiences of the student.Design. Prospective cohort study.Setting. Tygerberg Hospital, the Health Sciences Faculty of the University of Stellenbosch during a 15-week period from 4 February to 19 May 2002.Subjects. One hundred and thirty-six student interns (SIs), i.e. final-year medical students.Method. All SIs received a questionnaire and a letter motivating them to participate in the study and explaining the procedure. Regular class meetings enabled continuous motivation and ongoing updates. In the case of an incident during the 15-week period, the SI filled in the form and placed it in a sealed drop-off box.Outcome measures. Specific focus on the preceding events and the situation in which the incidents occurred (department, time of day, procedure performed, and whether the student was on call), exposure to HIV (patient's retroviral status), use of post-exposure prophylaxis (PEP) (whether used, when initiated), and the consequences of the exposure (emotional, on sexual behaviour during the window period, and on career choice).Results. During the 15-week period, 19 incidents were reported; the majority occurred while students were on call, almost half occurred after hours, and a disproportionate number occurred in three departments.Conclusions. Occupational blood exposure is a very real problem and poses a significant risk. SI suggestions should be considered in improving the prevention and management of such incidents

A cluster of high psychological and somatic symptoms in children with idiopathic scoliosis predicts persistent pain and analgesic use 1Â year after spine fusion

Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/146397/1/pan13467_am.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/146397/2/pan13467.pd

"Nothing is Whiter than White in this World": Child Sponsorship and the Geographies of Charity

In light of a scant, fragmentary geographical literature attending specifically to charity and charitable giving (cf. Bryson et al, 2002), this research presents an in-depth exploration of one particular (and highly popular) ‘charity’ mechanism- child sponsorship –by way of delineating a more coherent set of geographical understandings and sensibilities towards the topic. Using research carried out in the UK between 2011 and 2012 with both child sponsorship charities and ‘sponsors’, and drawing together an array of theoretical and conceptual resources from within geography and beyond, I seek to engage particularly with the ways in which charity is organised, promoted and practised; the spatial, relational ways in which charitable action is configured and performed, and the flows of ethical concern, embodied praxis and power which co-constitute it. As such, and mobilising ‘relational’ geographical work on networks and assemblages, I present an alternative reading of ‘charitable space’ which allows for its dynamic complexities to be more fully appreciated. Given my focus on child sponsorship, I set these interests within broader debates on the UK’s Third Sector, international development and humanitarian aid, particularly debates regarding neoliberalism and (post)colonialism. As such, the research also contributes to an emerging literature on Global North ‘development constituencies’ and their mobilisation (Baillie Smith, 2008; see also Smith, 2004; Desforges, 2004), as well as to well-established geographical literatures on voluntarism. I also foreground a focus on the dynamics of ‘faith-based’ giving, since the empirical landscape of child sponsorship displays a distinct orientation towards Christian modes of charitable organisation and action, though in complex, often blurry ways. In all, the work seeks to critically appraise and (where appropriate) disturb common narratives and assumptions used to apprehend charity in both popular and academic discourse, and offer instead a more critically attuned set of understandings which re-imagine charity in more enlivened ways.University of Exete

PCRPi, Presaging Critical Residues in Protein interfaces, a new computational tool to chart hot spots in protein interfaces

Protein–protein interactions (PPIs) are ubiquitous in Biology, and thus offer an enormous potential for the discovery of novel therapeutics. Although protein interfaces are large and lack defining physiochemical traits, is well established that only a small portion of interface residues, the so-called hot spot residues, contribute the most to the binding energy of the protein complex. Moreover, recent successes in development of novel drugs aimed at disrupting PPIs rely on targeting such residues. Experimental methods for describing critical residues are lengthy and costly; therefore, there is a need for computational tools that can complement experimental efforts. Here, we describe a new computational approach to predict hot spot residues in protein interfaces. The method, called Presaging Critical Residues in Protein interfaces (PCRPi), depends on the integration of diverse metrics into a unique probabilistic measure by using Bayesian Networks. We have benchmarked our method using a large set of experimentally verified hot spot residues and on a blind prediction on the protein complex formed by HRAS protein and a single domain antibody. Under both scenarios, PCRPi delivered consistent and accurate predictions. Finally, PCRPi is able to handle cases where some of the input data is either missing or not reliable (e.g. evolutionary information)

LMO2 and IL2RG synergize in thymocytes to mimic the evolution of SCID-X1 gene therapy-associated T-cell leukaemia

The SCID-X1 disease occurs in males that lack a functional X-linked gene encoding the interleukin 2 receptor subunit gamma (IL2RG) and thus are immuno-deficient (reviewed in Rochman et al.). Gene therapy has been a success in curing SCID-X1 in patients receiving autologous CD34+-bone marrow cells infected with retroviruses expressing IL2RG. This treatment protocol has, however, produced adverse T-cell effects where clonal T-cell leukaemias arose, and four have insertional mutagenesis of the T-cell oncogene LMO2. LMO2 is a T-cell oncogene first discovered via chromosomal translocations in T-cell acute leukaemia (T-ALL) (reviewed in Chambers and Rabbitts). It is unclear if the T-cell neoplasias in the SCID-X1 patients are simply due to insertional activation of the LMO2 gene or reflect synergy between LMO2 and IL2RG. Further, the recurrent involvement of LMO2 in SCID-X1 leukaemias is puzzling as other T-cell oncogenes (for example, TAL1/SCL, HOX11 and LYL1) might equally have been targets. This suggests that specific properties of LMO2 per se are required in these adverse events. The oncogenic potential of IL2RG itself also remains controversial. Although it causes T-cell lymphomas in mice transplanted with virally transduced haematopoetic stem cells, other studies have indicated that IL2RG is not an oncogene. Here we provide evidence that synergy is required between LMO2 and IL2RG proteins specifically in the T-cell lineage to elicit neoplasias and that additional mutations are required such as Notch1 mutations like those in human T-ALL

Genomic evidence of pre-invasive clonal expansion, dispersal and progression in bronchial dysplasia

The term ‘field cancerization’ is used to describe an epithelial surface that has a propensity to develop cancerous lesions, and in the case of the aerodigestive tract this is often as a result of chronic exposure to carcinogens in cigarette smoke 1, 2. The clinical endpoint is the development of multiple tumours, either simultaneously or sequentially in the same epithelial surface. The mechanisms underlying this process remain unclear; one possible explanation is that the epithelium is colonized by a clonal population of cells that are at increased risk of progression to cancer. We now address this possibility in a short case series, using individual genomic events as molecular biomarkers of clonality. In squamous lung cancer the most common genomic aberration is 3q amplification. We use a digital PCR technique to assess the clonal relationships between multiple biopsies in a longitudinal bronchoscopic study, using amplicon boundaries as markers of clonality. We demonstrate that clonality can readily be defined by these analyses and confirm that field cancerization occurs at a pre-invasive stage and that pre-invasive lesions and subsequent cancers are clonally related. We show that while the amplicon boundaries can be shared between different biopsies, the degree of 3q amplification and the internal structure of the 3q amplicon varies from lesion to lesion. Finally, in this small cohort, the degree of 3q amplification corresponds to clinical progression. Copyright © 2011 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd