Abstracts of the 24th international isotope society (UK group) symposium: synthesis and applications of labelled compounds 2015

The 24th annual symposium of the International Isotope Society's United Kingdom Group took place at the Møller Centre, Churchill College, Cambridge, UK on Friday 6th November 2015. The meeting was attended by 77 delegates from academia and industry, the life sciences, chemical, radiochemical and scientific instrument suppliers. Delegates were welcomed by Dr Ken Lawrie (GlaxoSmithKline, UK, chair of the IIS UK group). The subsequent scientific programme consisted of oral presentations, short ‘flash’ presentations in association with particular posters and poster presentations. The scientific areas covered included isotopic synthesis, regulatory issues, applications of labelled compounds in imaging, isotopic separation and novel chemistry with potential implications for isotopic synthesis. Both short‐lived and long‐lived isotopes were represented, as were stable isotopes. The symposium was divided into a morning session chaired by Dr Rebekka Hueting (University of Oxford, UK) and afternoon sessions chaired by Dr Sofia Pascu (University of Bath, UK) and by Dr Alan Dowling (Syngenta, UK). The UK meeting concluded with remarks from Dr Ken Lawrie (GlaxoSmithKline, UK).     

Thoughts of harming infants in depressed and nondepressed mothers.

BACKGROUND: Thoughts of harming the infant and other disturbing cognitions are frequently described in anecdotal reports on postpartum depression. These cognitions have not been examined empirically. METHODS: 100 clinically depressed mothers with a child under 3 years were evaluated and compared to a control group of 46 nondepressed mothers. RESULTS: 41% of depressed mothers compared to 7% of control mothers admitted to thoughts of harming their infant. Fear of being alone with the infant and inability to care for the infant were assessed only in depressed mothers and occurred less frequently. More than half of depressed mothers had problems in one of these three areas. CONCLUSIONS: Thoughts of harming the infant are common in depressed mothers. Demographic variables, psychosocial stressors and psychiatric variables do not help predict which mothers are likely to experience thoughts of harm or fear of being alone with the infant. These cognitive and affective disturbances may be one pathway by which maternal depression affects infants. LIMITATIONS: The control group was not given the full diagnostic interview. Consequently, the groups were not selected by identical procedures. Also fear of being alone with the infant and difficulty caring for the infant were not assessed in the control group.     

Pre-clinical validation of a novel alpha-7 nicotinic receptor radiotracer, [(3)H]AZ11637326: target localization, biodistribution and ligand occupancy in the rat brain

The alpha-7 neuronal nicotinic receptor is a novel pharmacological target for psychiatric and cognitive disorders. Selective radiotracer tools for pre-clinical receptor occupancy can facilitate the interpretation of the biological actions of small molecules at a target receptor. We discovered a high affinity nicotinic alpha-7 subtype-selective ligand, AZ11637326, with physical-chemical and pharmacokinetic properties suitable for an in vivo radioligand tool. [(3)H]AZ11637326 synthesis by tritiodehalogenation of the corresponding tribromide precursor yielded a high specific activity radiotracer with high affinity alpha-7 receptor binding in the rat hippocampus determined by autoradiography (Kd?=?0.2?nM). When [(3)H]AZ11637326 was administered to rats by intravenous bolus, rapid uptake was measured in the brain followed by a 3-4 fold greater specific binding in regions containing the alpha-7 receptor (frontal cortex, hippocampus, hypothalamus and midbrain) when compared to non-target regions (striatum and cerebellum). Systemic administration of the high affinity alpha-7 receptor antagonist, methyllycaconitine (MLA), or pretreatment with alpha-7 selective agonists (AR-R17779, PyrQTC, DBCO-4-POM, and DBCO-3-POM) significantly blocked the alpha-7 specific binding of [(3)H]AZ11637326 in the rat brain. The rank order of ligand ED(50) values for in vivo alpha-7 receptor occupancy in rat hippocampus was: DBCO-4-POM?>?DBCO-3-POM?～?MLA?>?PyrQTC?>?AR-R17779. The occupancy affinity shift was consistent with in vitro binding affinity in autoradiography. Our studies established the optimal conditions for [(3)H]AZ11637326 in vivo specific binding in the rat brain and support the use of [(3)H]AZ11637326 as a pre-clinical tool for assessment of novel alpha-7 compounds in drug discovery.     

Joint modeling of sensitivity and specificity

Sensitivity and specificity are two customary performance measures associated with medical diagnostic tests. Typically, they are modeled independently as a function of risk factors using logistic regression, which provides estimated functions for these probabilities. Change in these probabilities across levels of risk factors is of primary interest and the indirect relationship is often displayed using a receiver operating characteristic curve. We refer to this as analysis of 'first-order' behavior. Here, we consider what we refer to as 'second-order' behavior where we examine the stochastic dependence between the (random) estimates of sensitivity and specificity. To do so, we argue that a model for the four cell probabilities that determine the joint distribution of screening test result and outcome result is needed. Such a modeling induces sensitivity and specificity as functions of these cell probabilities. In turn, this raises the issue of a coherent specification for these cell probabilities, given risk factors, i.e. a specification that ensures that all probabilities calculated under it fall between 0 and 1. This leads to the question of how to provide models that are coherent and mechanistically appropriate as well as computationally feasible to fit, particularly with large data sets. The goal of this article is to illuminate these issues both algebraically and through analysis of a real data set.     

Match criteria for human cell line authentication: Where do we draw the line?

Continuous human cell lines have been used extensively as models for biomedical research. In working with these cell lines, researchers are often unaware of the risk of cross‐contamination and other causes of misidentification. To reduce this risk, there is a pressing need to authenticate cell lines, comparing the sample handled in the laboratory to a previously tested sample. The American Type Culture Collection Standards Development Organization Workgroup ASN‐0002 has developed a Standard for human cell line authentication, recommending short tandem repeat (STR) profiling for authentication of human cell lines. However, there are known limitations to the technique when applied to cultured samples, including possible genetic drift with passage. In our study, a dataset of 2,279 STR profiles from four cell banks was used to assess the effectiveness of the match criteria recommended within the Standard. Of these 2,279 STR profiles, 1,157 were grouped into sets of related cell lines—duplicate holdings, legitimately related samples or misidentified cell lines. Eight core STR loci plus amelogenin were used to unequivocally authenticate 98% of these related sets. Two simple match algorithms each clearly discriminated between related and unrelated samples, with separation between related samples at ≥80% match and unrelated samples at <50% match. A small degree of overlap was noted at 50–79% match, mostly from cell lines known to display variable STR profiles. These match criteria are recommended as a simple and effective way to interpret results from STR profiling of human cell lines.     

Nitric oxide in experimental aneurysm formation: early events and consequences of nitric oxide inhibition

Mounting evidence suggests that nitric oxide (NO) plays an important role in aneurysm pathogenesis. Nitric oxide synthase (NOS) expression, hemodynamic consequences of NO inhibition, and the effect of NO on matrix metalloproteinase (MMP) expression during aneurysm formation are unknown. In this study, a standard intraaortic elastase infusion rat model was used. Control animals received intraaortic elastase infusion and intraperitoneal saline injections. Experimental groups received intraaortic elastase infusion and intraperitoneal injections of aminoguanidine (500 mg/kg) or L-NAME (2 mg/kg or 10 mg/kg). Aortic diameter, blood pressure, and NO metabolites were measured at surgery and postoperative (POD) 7. A second series of rats were randomly infused with intraaortic elastase or saline and aortas were analyzed on POD 1, 3, and 7 with Western blotting for iNOS, eNOS, and MMP-9 expression. Infusion of elastase produced aneurysms (p <0.0001) in all rats. Inhibition of NO with aminoguanidine or L-NAME limited aneurysm expansion in all groups (p <0.05). Nitric oxide metabolites were increased (p <0.003) in control rats on POD 7. Arterial hypertension was present in all treated animals (p <0.05). Early up-regulation on POD 1 of iNOS (p <0.003) was noted in elastase-infused animals, but there was no iNOS expression with saline infusion. MMP-9 expression was present in both groups, with a significant increase in expression for elastase-infused animals noted on POD 7. iNOS expression is up-regulated early in experimental aneurysm formation, followed by increases in MMP-9 expression. Inhibition of NO limits aneurysmal expansion despite production of arterial hypertension.