A Forward-Design Approach to Increase the Production of Poly-3-Hydroxybutyrate in Genetically Engineered Escherichia coli

Biopolymers, such as poly-3-hydroxybutyrate (P(3HB)) are produced as a carbon store in an array of organisms and exhibit characteristics which are similar to oil-derived plastics, yet have the added advantages of biodegradability and biocompatibility. Despite these advantages, P(3HB) production is currently more expensive than the production of oil-derived plastics, and therefore, more efficient P(3HB) production processes would be desirable. In this study, we describe the model-guided design and experimental validation of several engineered P(3HB) producing operons. In particular, we describe the characterization of a hybrid phaCAB operon that consists of a dual promoter (native and J23104) and RBS (native and B0034) design. P(3HB) production at 24 h was around six-fold higher in hybrid phaCAB engineered Escherichia coli in comparison to E. coli engineered with the native phaCAB operon from Ralstonia eutropha H16. Additionally, we describe the utilization of non-recyclable waste as a low-cost carbon source for the production of P(3HB)

American Society for Echocardiography Software Suite for Verification and

The imminent balloting of the draft DICOM 3.0 module on ultrasound represents a major step forward in the progress of digital storage formats for echocardiography. In this paper, we discuss a software suite soon to be released into the public domain for reading, parsing and verifying DICOM 3.0 ultrasonic objects. The software is designed to allow the user to verify that a Jle consisting of digital ultrasound data, claiming DlCOM 3.0 conformance, does in fact adhere to the standard. A graphical user-interface for the software suite is also provided, allowing the playback of cin& mode images in real-time on standard computer hardware, and across a TCP/IP network

Scale Invariance in the Nonstationarity of Physiological Signals

We introduce a segmentation algorithm to probe temporal organization of heterogeneities in human heartbeat interval time series. We find that the lengths of segments with different local values of heart rates follow a power-law distribution. This scale-invariant structure is not a simple consequence of the long-range correlations present in the data. We also find that the differences in mean heart rates between consecutive segments display a common functional form, but with different parameters for healthy individuals and for patients with heart failure. This finding may provide information into the way heart rate variability is reduced in cardiac disease.Comment: 13 pages, 5 figures, corrected typo

Bladder spontaneous activity : influence of mild heating and inert injectables.

Overactive bladder (OAB) syndrome is associated with increased spontaneous contractions of the bladder wall, potentially mediated by the release of chemical agents from the urothelium. Reduction of these spontaneous contractions offers a means to alleviate the symptoms of OAB. I tested two novel approaches using pig bladder in vitro or ex vivo preparations; bladder wall heating or injection of inert bulking agents. In vitro, intact (mucosa + detrusor muscle) preparations were heated to 42, 46, or 50oC by a heating coil. Preparations of only detrusor muscle or mucosa were heated by changing superfusate temperature. Experiments were done to examine the role of heat-sensitive TRPV1 channels during heating by using TRPV1 antagonists. The effect of heating on urothelium ATP release was also measured. Possible changes to tissue structure were histologically assessed with haematoxylin & eosin or van Gieson staining. Inert bulking agents (Tyrode’s, polyethylene glycol or coaptite) were injected into the sub-mucosal space and their effect on spontaneous contractions also measured. Ex vivo experiments with perfused pig bladders recorded spontaneous pressure variations when perfusate temperature was increased to 42°C. Spontaneous contractions in intact preparations were reversibly reduced when heated to 42, 46, or 50oC; TRPV1 antagonists had no effect. Heating to 42oC did not affect mucosa or detrusor-only preparations, but at 50°C contractions were abolished. Similar effects were seen in ex vivo experiments when heated to 42°C. No changes to tissue integrity were observed at 42 or 46°C. At all temperatures urothelial ATP release was increased. Spontaneous contractions were reduced by all bulking agents, coaptite was the most effective. These novel findings suggest possible clinical approaches to treat the symptoms of OAB by reducing spontaneous contractions

Rapid prototyping platform for Saccharomyces cerevisiae using computer-aided genetic design enabled by parallel software and workcell platform development

Biofoundries have enabled the ability to automate the construction of genetic constructs using computer-aided design. In this study, we have developed the methodology required to abstract and automate the construction of yeast-compatible designs. We demonstrate the use of our in-house software tool, AMOS, to coordinate with design software, JMP, and robotic liquid handling platforms to successfully manage the construction of a library of 88 yeast expression plasmids. In this proof-of-principle study, we used three fluorescent genes as proxy for three enzyme coding sequences. Our platform has been designed to quickly iterate around a design cycle of four protein coding sequences per plasmid, with larger numbers possible with multiplexed genome integrations in Saccharomyces cerevisiae. This work highlights how developing scalable new biotechnology applications requires a close integration between software development, liquid handling robotics, and protocol development

Contractile effects and receptor analysis of adenosine-receptors in human detrusor muscle from stable and neuropathic bladders

To measure the relative transcription of adenosine receptor subtypes and the contractile effects of adenosine and selective receptor-subtype ligands on detrusor smooth muscle from patients with neuropathic overactive (NDO) and stable bladders and also from guinea-pigs. Contractile function was measured at 37°C in vitro from detrusor smooth muscle strips. Contractions were elicited by superfusate agonists or by electrical field stimulation. Adenosine-receptor (A1, A2A, A2B, A3) transcription was measured by RT-PCR. Adenosine attenuated nerve-mediated responses with equivalent efficacy in human and guinea-pig tissue (pIC50 3.65–3.86); the action was more effective at low (1–8 Hz) compared to high (20– 40 Hz) stimulation frequencies in human NDO and guinea-pig tissue. With guinea-pig detrusor the action of adenosine was mirrored by the A1/A2-agonist N-ethylcarboxamido adenosine (NECA), partly abolished in turn by the A2B-selectve antagonist alloxazine, as well as the A1-selective agonist N6- cyclopentyladenosine (CPA). With detrusor from stable human bladders the effects of NECA and CPA were much smaller than that of adenosine. Adenosine also attenuated carbachol contractures, but mirrored by NECA (in turn blocked by alloxazine) only in guinea-pig tissue. Adenosine receptor subtype transcription was measured in human detrusor and was similar in both groups, except reduced A2A levels in overactive bladder. Suppression of the carbachol contracture in human detrusor is independent of A-receptor activation, in contrast to an A2B-dependent action with guinea-pig tissue. Adenosine also reduced nerve-mediated contractions, by an A1- dependent action suppressing ATP neurotransmitter action