Thiamine is synthesized by a salvage pathway in Rhizobium leguminosarum bv. viciae strain 3841

In the absence of added thiamine, Rhizobium leguminosarum bv. viciae strain 3841 does not grow in liquid medium and forms only "pin" colonies on agar plates, which contrasts with the good growth of Sinorhizobium meliloti 1021, Mesorhizobium loti 303099, and Rhizobium etli CFN42. These last three organisms have thiCOGE genes, which are essential for de novo thiamine synthesis. While R. leguminosarum bv. viciae 3841 lacks thiCOGE, it does have thiMED. Mutation of thiM prevented formation of pin colonies on agar plates lacking added thiamine, suggesting thiamine intermediates are normally present. The putative functions of ThiM, ThiE, and ThiD are 4-methyl-5-(beta-hydroxyethyl) thiazole (THZ) kinase, thiamine phosphate pyrophosphorylase, and 4-amino-5-hydroxymethyl-2-methyl pyrimidine (HMP) kinase, respectively. This suggests that a salvage pathway operates in R. leguminosarum, and addition of HMP and THZ enabled growth at the same rate as that enabled by thiamine in strain 3841 but elicited no growth in the thiM mutant (RU2459). There is a putative thi box sequence immediately upstream of the thiM, and a gfp-mut3.1 fusion to it revealed the presence of a promoter that is strongly repressed by thiamine. Using fluorescent microscopy and quantitative reverse transcription-PCR, it was shown that thiM is expressed in the rhizosphere of vetch and pea plants, indicating limitation for thiamine. Pea plants infected by RU2459 were not impaired in nodulation or nitrogen fixation. However, colonization of the pea rhizosphere by the thiM mutant was impaired relative to that of the wild type. Overall, the results show that a thiamine salvage pathway operates to enable growth of Rhizobium leguminosarum in the rhizosphere, allowing its survival when thiamine is limiting.

A practical approach to a multi-level analysis with a sparse binary outcome within a large surgical trial

Objectives: To assess the potential source of variation that surgeon may add to patient outcome in a clinical trial of surgical procedures. Methods: Two large (n = 1380) parallel multicentre randomized surgical trials were undertaken to compare laparoscopically assisted hysterectomy with conventional methods of abdominal and vaginal hysterectomy; involving 43 surgeons. The primary end point of the trial was the occurrence of at least one major complication. Patients were nested within surgeons giving the data set a hierarchical structure. A total of 10% of patients had at least one major complication, that is, a sparse binary outcome variable. A linear mixed logistic regression model (with logit link function) was used to model the probability of a major complication, with surgeon fitted as a random effect. Models were fitted using the method of maximum likelihood in SAS((R)). Results: There were many convergence problems. These were resolved using a variety of approaches including; treating all effects as fixed for the initial model building; modelling the variance of a parameter on a logarithmic scale and centring of continuous covariates. The initial model building process indicated no significant 'type of operation' across surgeon interaction effect in either trial, the 'type of operation' term was highly significant in the abdominal trial, and the 'surgeon' term was not significant in either trial. Conclusions: The analysis did not find a surgeon effect but it is difficult to conclude that there was not a difference between surgeons. The statistical test may have lacked sufficient power, the variance estimates were small with large standard errors, indicating that the precision of the variance estimates may be questionable.

Graphical tracking systems revisited: a practical approach to computer scheduling in horticulture

Graphical tracking is a technique for crop scheduling where the actual plant state is plotted against an ideal target curve which encapsulates all crop and environmental characteristics. Management decisions are made on the basis of the position of the actual crop against the ideal position. Due to the simplicity of the approach it is possible for graphical tracks to be developed on site without the requirement for controlled experimentation. Growth models and graphical tracks are discussed, and an implementation of the Richards curve for graphical tracking described. In many cases, the more intuitively desirable growth models perform sub-optimally due to problems with the specification of starting conditions, environmental factors outside the scope of the original model and the introduction of new cultivars. Accurate specification for a biological model requires detailed and usually costly study, and as such is not adaptable to a changing cultivar range and changing cultivation techniques. Fitting of a new graphical track for a new cultivar can be conducted on site and improved over subsequent seasons. Graphical tracking emphasises the current position relative to the objective, and as such does not require the time consuming or system specific input of an environmental history, although it does require detailed crop measurement. The approach is flexible and could be applied to a variety of specification metrics, with digital imaging providing a route for added value. For decision making regarding crop manipulation from the observed current state, there is a role for simple predictive modelling over the short term to indicate the short term consequences of crop manipulation.

Regulation of L-alanine dehydrogenase in Rhizobium leguminosarum bv. viciae and its role in pea nodules

Alanine dehydrogenase (AldA) is the principal enzyme with which pea bacteroids synthesize alanine de novo. In free-living culture, AMA activity is induced by carboxylic acids (succinate, malate, and pyruvate), although the best inducer is alanine. Measurement of the intracellular concentration of alanine showed that AldA contributes to net alanine synthesis in laboratory cultures. Divergently transcribed from aldA is an AsnC type regulator, aldR. Mutation of aldR prevents induction of AldA activity. Plasmid-borne gusA fusions showed that aldR is required for transcription of both aldA and aldR; hence, AldR is autoregulatory. However, plasmid fusions containing the aldA-aldR intergenic region could apparently titrate out AldR, sometimes resulting in a complete loss of AldA enzyme activity. Therefore, integrated aldR::gusA and aldA::gusA fusions, as well as Northern blotting, were used to confirm the induction of aldA activity. Both aldA and aldR were expressed in the II/III interzone and zone III of pea nodules. Overexpression of aldA in bacteroids did not alter the ability of pea plants to fix nitrogen, as measured by acetylene reduction, but caused a large reduction in the size and dry weight of plants. This suggests that overexpression of aldA impairs the ability of bacteroids to donate fixed nitrogen that the plant can productively assimilate. We propose that the role of AldA may be to balance the alanine level for optimal functioning of bacteroid metabolism rather than to synthesize alanine as the sole product of N-2 reduction.

A practical low cost architecture for a MB-OFDM equalizer (ECMA-368)

The relative fast processing speed requirements in Wireless Personal Area Network (WPAN) consumer based products are often in conflict with their low power and cost requirements. In order to solve this conflict the efficiency and cost effectiveness of these products and the underlying functional modules become paramount. This paper presents a low-cost, simple, yet high performance solution for the receiver Channel Estimator and Equalizer for the Mutiband OFDM (MB-OFDM) system, particularly directed to the WiMedia Consortium Physical Later (ECMA-368) consumer implementation for Wireless-USB and Fast Bluetooth. In this paper, the receiver fixed point performance is measured and the results indicate excellent performance compared to the current literature(1).

Robot-human interaction - practical experiments with a cyborg

This paper presents results to indicate the potential applications of a direct connection between the human nervous system and a computer network. Actual experimental results obtained from a human subject study are given, with emphasis placed on the direct interaction between the human nervous system and possible extra-sensory input. An brief overview of the general state of neural implants is given, as well as a range of application areas considered. An overall view is also taken as to what may be possible with implant technology as a general purpose human-computer interface for the future.

Combining Human and Machine Brains - Practical systems in information & control

In this paper a look is taken at how the use of implant technology can be used to either increase the range of the abilities of a human and/or diminish the effects of a neural illness, such as Parkinson's Disease. The key element is the need for a clear interface linking the human brain directly with a computer. The area of interest here is the use of implant technology, particularly where a connection is made between technology and the human brain and/or nervous system. Pilot tests and experimentation are invariably carried out apriori to investigate the eventual possibilities before human subjects are themselves involved. Some of the more pertinent animal studies are discussed here. The paper goes on to describe human experimentation, in particular that carried out by the author himself, which led to him receiving a neural implant which linked his nervous system bi-directionally with the internet. With this in place neural signals were transmitted to various technological devices to directly control them. In particular, feedback to the brain was obtained from the fingertips of a robot hand and ultrasonic (extra) sensory input. A view is taken as to the prospects for the future, both in the near term as a therapeutic device and in the long term as a form of enhancement.

A practical method for testing high-speed networking hardware architectures

This paper deals with the key issues encountered in testing during the development of high-speed networking hardware systems by documenting a practical method for "real-life like" testing. The proposed method is empowered by modern and commonly available Field Programmable Gate Array (FPGA) technology. Innovative application of standard FPGA blocks in combination with reconfigurability are used as a back-bone of the method. A detailed elaboration of the method is given so as to serve as a general reference. The method is fully characterised and compared to alternatives through a case study proving it to be the most efficient and effective one at a reasonable cost.