Isolation and structural determination of non-racemic tertiary cathinone derivatives

The racemic tertiary cathinones N,N-dimethylcathinone (1), N,N-diethylcathinone (2) and 2-(1-pyrrolidinyl)-propiophenone (3) have been prepared in reasonable yield and characterized using NMR and mass spectroscopy. HPLC indicates that these compounds are isolated as the anticipated racemic mixture. These can then be co-crystallized with (+)-O,O′-di-p-toluoyl-d-tartaric, (+)-O,O′-dibenzoyl-d-tartaric and (-)-O,O′-dibenzoyl-l-tartaric acids giving the single enantiomers S and R respectively of 1, 2 and 3, in the presence of sodium hydroxide through a dynamic kinetic resolution. X-ray structural determination confirmed the enantioselectivity. The free amines could be obtained following basification and extraction. In methanol these are reasonably stable for the period of several hours, and their identity was confirmed by HPLC and CD spectroscopy.

Complete framework for efficient characterisation of non-Markovian processes

A non-Markovian process is one that retains `memory' of its past. A systematic understanding of these processes is necessary to fully describe and harness a vast range of complex phenomena; however, no such general characterisation currently exists. This long-standing problem has hindered advances in understanding physical, chemical and biological processes, where often dubious theoretical assumptions are made to render a dynamical description tractable. Moreover, the methods currently available to treat non-Markovian quantum dynamics are plagued with unphysical results, like non-positive dynamics. Here we develop an operational framework to characterise arbitrary non-Markovian quantum processes. We demonstrate the universality of our framework and how the characterisation can be rendered efficient, before formulating a necessary and sufficient condition for quantum Markov processes. Finally, we stress how our framework enables the actual systematic analysis of non-Markovian processes, the understanding of their typicality, and the development of new master equations for the effective description of memory-bearing open-system evolution.

Tyrosine kinase inhibitor insensitivity of non-cycling CD34+ human acute myeloid leukaemia cells with FMS-like tyrosine kinase 3 mutations.

The efficacy of tyrosine kinase (TK) inhibitors on non-cycling acute myeloid leukaemia (AML) cells, previously shown to have potent tumourigenic potential, is unknown. This pilot study describes the first attempt to characterize non-cycling cells from a small series of human FMS-like tyrosine kinase 3 (FLT3) mutation positive samples. CD34+ AML cells from patients with FLT3 mutation positive AML were cultured on murine stroma. In expansion cultures, non-cycling cells were found to retain CD34+ expression in contrast to dividing cells. Leukaemic gene rearrangements could be detected in non-cycling cells, indicating their leukaemic origin. Significantly, the FLT3-internal tandem duplication (ITD) mutation was found in the non-cycling fraction of four out of five cases. Exposure to the FLT3-directed inhibitor TKI258 clearly inhibited the growth of AML CD34+ cells in short-term cultures and colony-forming unit assays. Crucially, non-cycling cells were not eradicated, with the exception of one case, which exhibited exquisite sensitivity to the compound. Moreover, in longer-term cultures, TKI258-treated non-cycling cells showed no growth impairment compared to treatment-naive non-cycling cells. These findings suggest that non-cycling cells in AML may constitute a disease reservoir that is resistant to TK inhibition. Further studies with a larger sample size and other inhibitors are warranted.

Refining the diagnosis and EGFR status of non-small cell lung carcinoma in biopsy and cytologic material, using a panel of mucin staining, TTF-1, cytokeratin 5/6, and P63, and EGFR mutation analysis.

INTRODUCTION: The dichotomization of non-small cell carcinoma (NSCLC) subtype into squamous (SQCC) and adenocarcinoma (ADC) has become important in recent years and is increasingly required with regard to management. The aim of this study was to determine the utility of a panel of commercially available antibodies in refining the diagnosis on small biopsies and also to determine whether cytologic material is suitable for somatic EGFR genotyping in a prospectively analyzed series of patients undergoing investigation for suspected lung cancer. METHODS: Thirty-two consecutive cases of NSCLC were first tested using a panel comprising cytokeratin 5/6, P63, thyroid transcription factor-1, 34betaE12, and a D-PAS stain for mucin, to determine their value in refining diagnosis of NSCLC. After this test phase, two further pathologists independently reviewed the cases using a refined panel that excluded 34betaE12 because of its low specificity for SQCC, and refinement of diagnosis and concordance were assessed. Ten cases of ADC, including eight derived from cytologic samples, were sent for EGFR mutation analysis. RESULTS: There was refinement of diagnosis in 65% of cases of NSCLC to either SQCC or ADC in the test phase. This included 10 of 13 cases where cell pellets had been prepared from transbronchial needle aspirates. Validation by two further pathologists with varying expertise in lung pathology confirmed increased refinement and concordance of diagnosis. All samples were adequate for analysis, and they all showed a wild-type EGFR genotype. CONCLUSION: A panel comprising cytokeratin 5/6, P63, thyroid transcription factor-1, and a D-PAS stain for mucin increases diagnostic accuracy and agreement between pathologists when faced with refining a diagnosis of NSCLC to SQCC or ADC. These small samples, even cell pellets derived from transbronchial needle aspirates, seem to be adequate for EGFR mutation analysis.

A Loop-mediated Isothermal Amplification Method for Rapid Direct Detection and Differentiation of Non-pathogenic and Verocytotoxigenic Escherichia coli in Beef and Bovine Faeces

The aim of this study was to develop a multiplex loop-mediated isothermal amplification (LAMP) method capable of detecting Escherichia coli generally and verocytotoxigenic E. coli (VTEC) specifically in beef and bovine faeces. The LAMP assay developed was highly specific (100%) and able to distinguish between E. coli and VTEC based on the amplification of the phoA, and stx1 and/or stx2 genes, respectively. In the absence of an enrichment step, the limit of detection 50% (LOD50) of the LAMP assay was determined to be 2.83, 3.17 and 2.83-3.17 log CFU/g for E. coli with phoA, stx1 and stx2 genes, respectively, when artificially inoculated minced beef and bovine faeces were tested. The LAMP calibration curves generated with pure cultures, and spiked beef and faeces, suggested that the assay had good quantification capability. Validation of the assay, performed using retail beef and bovine faeces samples, demonstrated good correlation between counts obtained by the LAMP assay and by a conventional culture method, but suggested the possibility of false negative LAMP results for 12.5-14.7% of samples tested. The multiplex LAMP assay developed potentially represents a rapid alternative to culture for monitoring E.coli levels in beef or faeces and it would provide additional information on the presence of VTEC. However, some further optimisation is needed to improve detection sensitivity.

Fixed-boundary constituencies and the principle of equal representation in Ireland

The process of constituency boundary revision in Ireland, designed to satisfy what is perceived as a rigid requirement that a uniform deputy-population ratio be maintained across constituencies, has traditionally consumed a great deal of the time of politicians and officials. For almost two decades after a High Court ruling in 1961, the process was a political one, was highly contentious, and was marked by serious allegations of ministerial gerrymandering. The introduction in 1979 of constituency commissions made up of officials neutralised, for the most part, charges that the system had become too politicised, but it continued the process of micro-management of constituency boundaries. This article suggests that the continuing problems caused by this system – notably, the permanently changing nature of constituency boundaries and resulting difficulties of geographical identification – could be resolved by reversion to the procedure that is normal in proportional representation systems: periodic post-census allocation of seats to constituencies whose boundaries are based on those of recognised local government units and which are stable over time. This reform, replacing the principle of redistricting by the principle of reapportionment, would result in more recognisable constituencies, more predictable boundary trajectories over time, and a more efficient, fairer, and speedier process of revision.

Conclusion: patterns of non-territorial autonomy

This article seeks to generalise about the significance of non-territorial autonomy as a mechanism for the management of ethnic conflict on the basis of a set of case studies covering the Ottoman empire and its successor states, the Habsburg monarchy, the Jewish minorities of Europe, interwar Estonia, contemporary Belgium, and two indigenous peoples, the Sami in Norway and Maori in New Zealand. It begins by assessing the extent to which the spatial distribution of ethnonational communities determined the range of autonomy options available—whether these might be territorial or whether only non-territorial autonomy would be realistic. The article continues with an assessment of the significance of ‘autonomy’ in circumstances where the institutions with which it is associated enjoy a non-territorial rather than a territorial writ. It concludes by suggesting that in almost all cases where autonomy is extended to a minority within a state this is exercised on a territorial basis, and that in many cases of non-territorial autonomy, or national–cultural autonomy, the powers assumed by the ‘autonomous’ institutions are substantially symbolic. It argues that notwithstanding the limited empirical evidence for the existence of non-territorial autonomy, this device should not be written off at a normative level.