Efficient delivery of Cre-recombinase to neurons in vivo and stable transduction of neurons using adeno-associated and lentiviral vectors

BACKGROUND: Inactivating genes in vivo is an important technique for establishing their function in the adult nervous system. Unfortunately, conventional knockout mice may suffer from several limitations including embryonic or perinatal lethality and the compensatory regulation of other genes. One approach to producing conditional activation or inactivation of genes involves the use of Cre recombinase to remove loxP-flanked segments of DNA. We have studied the effects of delivering Cre to the hippocampus and neocortex of adult mice by injecting replication-deficient adeno-associated virus (AAV) and lentiviral (LV) vectors into discrete regions of the forebrain. RESULTS: Recombinant AAV-Cre, AAV-GFP (green fluorescent protein) and LV-Cre-EGFP (enhanced GFP) were made with the transgene controlled by the cytomegalovirus promoter. Infecting 293T cells in vitro with AAV-Cre and LV-Cre-EGFP resulted in transduction of most cells as shown by GFP fluorescence and Cre immunoreactivity. Injections of submicrolitre quantities of LV-Cre-EGFP and mixtures of AAV-Cre with AAV-GFP into the neocortex and hippocampus of adult Rosa26 reporter mice resulted in strong Cre and GFP expression in the dentate gyrus and moderate to strong labelling in specific regions of the hippocampus and in the neocortex, mainly in neurons. The pattern of expression of Cre and GFP obtained with AAV and LV vectors was very similar. X-gal staining showed that Cre-mediated recombination had occurred in neurons in the same regions of the brain, starting at 3 days post-injection. No obvious toxic effects of Cre expression were detected even after four weeks post-injection. CONCLUSION: AAV and LV vectors are capable of delivering Cre to neurons in discrete regions of the adult mouse brain and producing recombination

In-house Preparation of Examination Papers using troff, tbl, and eqn.

Starting in December 1982 the University of Nottingham decided to phototypeset almost all of its examination papers `in house' using the troff, tbl and eqn programs running under UNIX. This tutorial lecture highlights the features of the three programs with particular reference to their strengths and weaknesses in a production environment. The following issues are particularly addressed: Standards -- all three software packages require the embedding of commands and the invocation of pre-written macros, rather than `what you see is what you get'. This can help to enforce standards, in the absence of traditional compositor skills. Hardware and Software -- the requirements are analysed for an inexpensive preview facility and a low-level interface to the phototypesetter. Mathematical and Technical papers -- the fine-tuning of eqn to impose a standard house style. Staff skills and training -- systems of this kind do not require the operators to have had previous experience of phototypesetting. Of much greater importance is willingness and flexibility in learning how to use computer systems.

Modelling and analysing human behaviour in a department store using discrete event and agent based simulation

Discrete Event Simulation (DES) is a very popular simulation technique in Operational Research. Recently, there has been the emergence of another technique, namely Agent Based Simulation (ABS). Although there is a lot of literature relating to DES and ABS, we have found less that focuses on exploring the capabilities of both in tackling human behaviour issues. In order to understand the gap between these two simulation techniques, therefore, our aim is to understand the distinctions between DES and ABS models with the real world phenomenon in modelling and simulating human behaviour. In achieving the aim, we have carried out a case study at a department store. Both DES and ABS models will be compared using the same problem domain which is concerning on management policy in a fitting room. The behaviour of staffs while working and customers’ satisfaction will be modelled for both models behaviour understanding.