Production of Specialized Transformation Vectors for the Production of Biodegradable Plastics in Transgenic Arabidopsis and Oil Palm

Polyhydroxyalkanoates (PHAs), such as polyhydroxybutyrate (PHB) and polyhydroxybutyrate-co-hydroxyvalerate (PHBV) are bacterial polyesters, which can be used to produce biodegradable products. Since the mass production of PHAs in bacteria via fermentation is expensive, the production of PHAs in plants may be an attractive alternative. The production of PHB in plants required genetic engineering of phbA, phbB and phbC genes of Ralstonia eutropha, whereas, the bktB, phbB, phbC genes of R. eutropha and tdcB gene of Escherichia coli were required for PHBV production. In this study, each of these gene was fused with the transit peptide (Tp) of oil palm acyl-carrier-protein (ACP), and driven by the oil palm leaf-specific promoter (LSP1), for targeting into the plastids of leaf cells. In total, four transformation vectors, pLSP15 (PHB) and pLSP20 (PHBV), pLSP13 (PHB) and pLSP23 (PHBV) were constructed for the transformation of Arabidopsis and oil palm, respectively. Each vector contained the phosphinothricin acetyltransferase gene (Bar) driven by CaMV35S promoter in pLSP15 and pLSP20, and ubiquitin promoter in pLSP13 and pLSP23, as plant selectable marker. Matrix attachment region of tobacco (RB7MAR) was also included, to stabilize the transgene expression and to minimize gene silencing due to positional effects. Restriction enzymes, polymerase chain reaction (PCR) and DNA sequencing were used to verify all the constructed vectors. Arabidopsis transformation produced T1 transgenic Arabidopsis plants with normal phenotypes at a transformation efficiency of 0.2%~1.0%. PCR and Southern analyses were used to confirm the insertion of the transgenes. Nile blue A staining of these T1 plants demonstrated the accumulation of PHB granules in the leaf. The initial screening of Basta-resistant oil palm embryogenic calli transformed with pLSP13 using PCR demonstrated the presence of Bar and PHB genes in transformed oil palm

Transgenic plants producing polyhydroxyalkanoates

Currently, the polyhydroxybutyrate (PHB) copolymer, polyhydroxy-co-valerate (PHBV) is being produced commercially using a two-stage glucose / propionate fed batch fermentation process using Ralstonia eutropha. The economics of the manufacturing process are still a major barrier to the widespread use of polyhydroxyalkanoates (PHAs) and intensive efforts are being made to reduce the cost of production by means of bioprocess design and metabolic engineering of production strains. However, despite these improvements, the production costs are still high compared to petroleum-derived commodity plastics. An alternative strategy for lowering production costs that has been proposed is to develop transgenic plants that produce PHAs. This strategy is considerably cheaper because the PHAs production from plants does not require expensive fermentation equipment and processing facilities

Dynamic Increase in Corticomuscular Coherence during Bilateral, Cyclical Ankle Movements

In humans, the midline primary motor cortex is active during walking. However, the exact role of such cortical participation is unknown. To delineate the role of the primary motor cortex in walking, we examined whether the primary motor cortex would activate leg muscles during movements that retained specific requirements of walking (i.e., locomotive actions). We recorded electroencephalographic and electromyographic signals from 15 healthy, young men while they sat and performed bilateral, cyclical ankle movements. During dorsiflexion, near-20-Hz coherence increased cyclically between the midline primary motor cortex and the co-contracting antagonistic pair (i.e., tibialis anterior and medial gastrocnemius muscles) in both legs. Thus, we have shown that dynamic increase in corticomuscular coherence, which has been observed during walking, also occurs during simple bilateral cyclical movements of the feet. A possible mechanism for such coherence is corticomuscular communication, in which the primary motor cortex participates in the control of movement. Furthermore, because our experimental task isolated certain locomotive actions, the observed coherence suggests that the human primary motor cortex may participate in these actions (i.e., maintaining a specified movement frequency, bilaterally coordinating the feet, and stabilizing the posture of the feet). Additional studies are needed to identify the exact cortical and subcortical interactions that cause corticomuscular coherence and to further delineate the functional role of the primary motor cortex during bilateral cyclical movements such as walking

Construction of PHB and PHBV multiple-gene vectors driven by an oil palm leaf-specific promoter

One of the targets in oil palm genetic engineering programme is the production of polyhydroxybutyrate (PHB) and polyhydroxybutyrate-co-valerate (PHBV) in the oil palm leaf tissues. Production of PHB requires the use of phbA (β-ketothiolase type A), phbB (acetoacetyl-CoA reductase) and phbC (PHB synthase) genes of Ralstonia eutropha, whereas bktB (β-ketothiolase type B), phbB, phbC genes of R. eutropha and tdcB (threonine dehydratase) gene of Escherichia coli were used for PHBV production. Each of these genes was fused with a transit peptide (Tp) of oil palm acyl-carrier-protein (ACP) gene, driven by an oil palm leaf-specific promoter (LSP1) to genetically engineer the PHB/PHBV pathway to the plastids of the leaf tissues. In total, four transformation vectors, designated pLSP15 (PHB) and pLSP20 (PHBV), and pLSP13 (PHB) and pLSP23 (PHBV), were constructed for transformation in Arabidopsis thaliana and oil palm, respectively. The phosphinothricin acetyltransferase gene (bar) driven by CaMV35S promoter in pLSP15 and pLSP20, and ubiquitin promoter in pLSP13 and pLSP23 were used as the plant selectable markers. Matrix attachment region of tobacco (RB7MAR) was also included in the vectors to stabilize the transgene expression and to minimize silencing due to positional effect. Restriction digestion, PCR amplification and/or sequencing were carried out to ensure sequence integrity and orientation

What is the role body sway deviation and body sway velocity play in postural stability in older adults?

Summary: This cross sectional study focused on how Postural Stability (PS) indicators: body sway deviation (BSD) and body sway velocity (BSV), change with age and their association with levels of social and physical activity. Observational study: 80 older adults (aged: 60–96) were purposefully recruited from two sources: the University of the Third Age (TAU) (n = 35) and a residential care home (CH) (n = 45). Differences in the indicators of PS, approximated through Centre of Pressure (COP) measurements, were assessed by the Romberg Stance Test (Test A) subsequently repeated on 10 cm foam surface (Test B), using a Kistler® Dynamometric Platform. The RCH Group was older, had higher BMI and was less socially and physically active, showed more body sway in all indicators compared to TAU group. For all participants body sway velocity (BSV) was significantly correlated with age. The strength of correlation of body sway deviation (BSD) with age was also significant but not as strong. The findings indicate in line with previous studies that deterioration in BSV is associated with poor PS more than deterioration in BSD

On the exchange of intersection and supremum of sigma-fields in filtering theory

We construct a stationary Markov process with trivial tail sigma-field and a nondegenerate observation process such that the corresponding nonlinear filtering process is not uniquely ergodic. This settles in the negative a conjecture of the author in the ergodic theory of nonlinear filters arising from an erroneous proof in the classic paper of H. Kunita (1971), wherein an exchange of intersection and supremum of sigma-fields is taken for granted.Comment: 20 page

The MVGC multivariate Granger causality toolbox: a new approach to Granger-causal inference

Background: Wiener-Granger causality (“G-causality”) is a statistical notion of causality applicable to time series data, whereby cause precedes, and helps predict, effect. It is defined in both time and frequency domains, and allows for the conditioning out of common causal influences. Originally developed in the context of econometric theory, it has since achieved broad application in the neurosciences and beyond. Prediction in the G-causality formalism is based on VAR (Vector AutoRegressive) modelling. New Method: The MVGC Matlab c Toolbox approach to G-causal inference is based on multiple equivalent representations of a VAR model by (i) regression parameters, (ii) the autocovariance sequence and (iii) the cross-power spectral density of the underlying process. It features a variety of algorithms for moving between these representations, enabling selection of the most suitable algorithms with regard to computational efficiency and numerical accuracy. Results: In this paper we explain the theoretical basis, computational strategy and application to empirical G-causal inference of the MVGC Toolbox. We also show via numerical simulations the advantages of our Toolbox over previous methods in terms of computational accuracy and statistical inference. Comparison with Existing Method(s): The standard method of computing G-causality involves estimation of parameters for both a full and a nested (reduced) VAR model. The MVGC approach, by contrast, avoids explicit estimation of the reduced model, thus eliminating a source of estimation error and improving statistical power, and in addition facilitates fast and accurate estimation of the computationally awkward case of conditional G-causality in the frequency domain. Conclusions: The MVGC Toolbox implements a flexible, powerful and efficient approach to G-causal inference. Keywords: Granger causality, vector autoregressive modelling, time series analysi