Binding mechanism of affinity ligands for purification of plasmid DNA

Plasmid DNA for therapeutic and vaccination purposes must be highly purified. The high selectivity of affinity chromatography makes it ideal for the isolation of pDNA from complex biological feed stocks. Affinity chromatography makes use of the biological function and/or individual chemical structure of the interacting molecules. However, the success of any affinity purification protocol is dependent on the availability of suitable ligands. In this study, surface plasmon resonance (SPR) based Biacore system has been employed for the detection and quantification of the binding between lac operon (lacO) sequence contained in a pDNA and synthetic peptides based on the DNA-binding domain of the lac repressor protein, lad. The equilibrium dissociation constant (K D) and association and dissociation rate constants (ka, kd) for the interaction between plasmid DNA and designed peptides were determined.

The applicability of the Precaution Adoption Process Model for understanding self-regulatory driving behaviour among older drivers

1. Introduction The success of self-regulation, in terms of enhancing older drivers’ safety and maintaining their mobility, depends largely upon older drivers’ awareness of the declines in their driving abilities. Therefore, interventions targeted at increasing older drivers’ safety should aim to enhance their awareness of their physical, sensory and cognitive limitations. Moreover, previous research suggests that driving behaviour change may occur through stages and that interventions and feedback may be perceived differently at each stage. 2. Study aims To further understand the process of driving self-regulation among older adults by exploring their perceptions and experiences of self-regulation, using the PAPM as a framework. To investigate the possible impact of feedback on their driving on their decision making process. 3. Methodology Research tool: Qualitative focus groups (n=5 sessions) Recruitment: Posters, media, newspaper advertisement and emails Inclusion criteria: Aged 70 or more, English-speaking, current drivers Participants: Convenience sample of 27 men and women aged 74 to 90 in the Sunshine Coast and Brisbane city, Queensland, Australia. 4. Analysis Thematic analysis was conducted following the process outlined by Braun and Clarke (2006) to identify, analyse and report themes within the data. Four main themes were identified.

Cytokine expression and secretion by skeletal muscle cells : regulatory mechanisms and exercise effects

Cytokines are important mediators of various aspects of health and disease, including appetite, glucose and lipid metabolism, insulin sensitivity, skeletal muscle hypertrophy and atrophy. Over the past decade or so, considerable attention has focused on the potential for regular exercise to counteract a range of disease states by modulating cytokine production. Exercise stimulates moderate to large increases in the circulating concentrations of interleukin (IL)-6, IL-8, IL-10, IL-1 receptor antagonist, granulocyte-colony stimulating factor, and smaller increases in tumor necrosis factor-α, monocyte chemotactic protein-1, IL-1β, brain-derived neurotrophic factor, IL-12p35/p40 and IL-15. Although many of these cytokines are also expressed in skeletal muscle, not all are released from skeletal muscle into the circulation during exercise. Conversely, some cytokines that are present in the circulation are not expressed in skeletal muscle after exercise. The reasons for these discrepant cytokine responses to exercise are unclear. In this review, we address these uncertainties by summarizing the capacity of skeletal muscle cells to produce cytokines, analyzing other potential cellular sources of circulating cytokines during exercise, and discussing the soluble factors and intracellular signaling pathways that regulate cytokine synthesis (e.g., RNA-binding proteins, microRNAs, suppressor of cytokine signaling proteins, soluble receptors).

Is climate change litigation an effective strategy for promoting greater action to address climate change? What other legal mechanisms might be appropriate?

In recent times a widespread consensus on the reality and gravity of anthropogenic climate change has emerged. Perceived inadequacies in the Australian government’s legal and policy responses to climate change issues have resulted in environmental activists increasingly turning to the courts as a strategy to promote greater action to address adverse climate impacts. The efficacy of this strategy for achieving climate goals is limited by the time and expense of litigating, the restrictions inherent in environmental law administrative challenges, and the possibility that judicial decisions may be overruled by the legislature. To date, climate change litigation in Australia has met with varied success, yet its significance extends beyond the court room as an important mechanism for raising public, political and commercial awareness about climate change issues. Ultimately, however, the types of far-reaching changes needed to mitigate and manage adverse climate impacts require strong regulatory backing. The most effective approach to addressing the complex challenges posed by climate change is a coordinated suite of regulatory measures spearheaded by the Federal Government.

Reduced intensity conditioning for allogeneic hematopoietic stem cell transplant determines the kinetics of acute Graft-Versus-Host Disease

Background Preparative myeloablative conditioning regimens for allogeneic hematopoietic stem-cell transplantation (HSCT) may control malignancy and facilitate engraftment but also contribute to transplant related mortality, cytokine release, and acute graft-versus-host disease (GVHD). Reduced intensity conditioning (RIC) regimens have decreased transplant related mortality but the incidence of acute GVHD, while delayed, remains unchanged. There are currently no in vivo allogeneic models of RIC HSCT, limiting studies into the mechanism behind RIC-associated GVHD. Methods We developed two RIC HSCT models that result in delayed onset GVHD (major histocompatibility complex mismatched (UBI-GFP/BL6 [H-2b]→BALB/c [H-2d]) and major histocompatibility complex matched, minor histocompatibility mismatched (UBI-GFP/BL6 [H-2b]→BALB.B [H-2b])) enabling the effect of RIC on chimerism, dendritic cell (DC) chimerism, and GVHD to be investigated. Results In contrast with myeloablative conditioning, we observed that RIC-associated delayed-onset GVHD is characterized by low production of tumor necrosis factor-α, maintenance of host DC, phenotypic DC activation, increased T-regulatory cell numbers, and a delayed emergence of activated donor DC. Furthermore, changes to the peritransplant milieu in the recipient after RIC lead to the altered activation of DC and the induction of T-regulatory responses. Reduced intensity conditioning recipients suffer less early damage to GVHD target organs. However, as donor cells engraft, activated donor DC and rising levels of tumor necrosis factor-α are associated with a later onset of severe GVHD. Conclusions Delineating the mechanisms underlying delayed onset GVHD in RIC HSCT recipients is vital to improve the prediction of disease onset and allow more targeted interventions for acute GVHD.

A recurrent network in the lateral amygdala: A mechanism for temporal coincidence detection

The dorsal lateral amygdala (LAd) is a vital nucleus for the formation of associations between aversive unconditioned stimuli (US) and neutral stimuli, such as auditory tones, which can become conditioned (CS) to the US through temporal pairing. Important aspects of CS-US associations are believed to occur within the LAd, however relatively little is known about the temporal behavior of local LAd networks. Information about the CS and US enters the LA via a rapid and direct thalamic input and a longer latency cortical path...

Different correlation of Sphingosine-1-Phosphate receptor 1 with receptor activator of nuclear factor kappa B ligand and regulatory T cells in rat periapical lesions

Introduction Sphingosine-1-phosphate receptor 1 (S1P1) is crucial for regulation of immunity and bone metabolism. This study aimed to investigate the expression of S1P1 in rat periapical lesions and its relationship with receptor activator of nuclear factor kappa B ligand (RANKL) and regulatory T (Treg) cells. Methods Periapical lesions were induced by pulp exposure in the first lower molars of 55 Wistar rats. Thirty rats were killed on days 0, 7, 14, 21, 28, and 35, and their mandibles were harvested for x-ray imaging, micro–computed tomography scanning, histologic observation, immunohistochemistry, enzyme histochemistry, and double immunofluorescence analysis. The remaining 25 rats were killed on days 0, 14, 21, 28, and 35, and mandibles were harvested for flow cytometry. Results The volume and area of the periapical lesions increased from day 0 to day 21 and then remained comparably stable after day 28. S1P1-positive cells were observed in the inflammatory periapical regions; the number of S1P1-positive cells peaked at day 14 and then decreased from day 21 to day 35. The distribution of S1P1-positive cells was positively correlated with the dynamics of RANKL-positive cells but was negatively correlated with that of Treg cells. Conclusions S1P1 expression was differentially correlated with RANKL and Treg cell infiltration in the periapical lesions and is therefore a contributing factor to the pathogenesis of such lesions.

Normative requirements for regulatory compliance: An abstract formal framework

By definition, regulatory rules (in legal context called norms) intend to achieve specific behaviour from business processes, and might be relevant to the whole or part of a business process. They can impose conditions on different aspects of process models, e.g., control-flow, data and resources etc. Based on the rules sets, norms can be classified into various classes and sub-classes according to their effects. This paper presents an abstract framework consisting of a list of norms and a generic compliance checking approach on the idea of (possible) execution of processes. The proposed framework is independent of any existing formalism, and provides a conceptually rich and exhaustive ontology and semantics of norms needed for business process compliance checking. The possible uses of the proposed framework include to compare different compliance management frameworks (CMFs).

Property law reforms in Queensland: Is the regulatory burden on land transactions reduced?

The reduction of unnecessary regulation was a clear policy objective of the Queensland government during 2014. In the area of property sales significant reforms were introduced from 1 December 2014. This article examines the key aspects of these reforms and whether there has been a reduction in red tape for sellers and buyers of land.

Cooperative choice and its framing effect under threshold uncertainty in a provision point mechanism

This paper explores how threshold uncertainty affects cooperative behaviors in the provision of public goods and the prevention of public bads. The following facts motivate our study. First, environmental (resource) problems are either framed as public bads prevention or public goods provision. Second, the occurrence of these problems is characterized by thresholds that are interchangeably represented as "nonconvexity," "bifurcation," "bi-stability," or "catastrophes." Third, the threshold location is mostly unknown. We employ a provision point mechanism with threshold uncertainty and analyze the responses of cooperative behaviors to uncertainty and to the framing for each type of social preferences categorized by a value orientation test. We find that aggregate framing effects are negligible, although the response to the frame is the opposite depending on the type of social preferences. "Cooperative" subjects become more cooperative in negative frames than in positive frames, whereas "individualistic" subjects are less cooperative in negative frames than in positive ones. This finding implies that the insignificance of aggregate framing effects arises from behavioral asymmetry. We also find that the percentage of cooperative choices non-monotonically varies with the degree of threshold uncertainty, irrespective of framing and value orientation. Specifically, the degree of cooperation is highest at intermediate levels of threshold uncertainty and decreases as the uncertainty becomes sufficiently large.

Effective intercalation of sodium dodecylsulfate (SDS) into hydrocalumite: Mechanism discussion via near-infrared and mid-infrared investigations

The intercalation of an anionic surfactant, sodium dodecylsulfate (SDS), into hydrocalumite (CaAl-LDH-Cl) was investigated in this study. To understand the intercalation behavior, X-ray diffraction (XRD), mid-infrared spectroscopy (MIR), near-infrared spectroscopy (NIR) and scanning electron microscopy (SEM) were undertaken. The near-infrared spectra indicated a special spectral range from 6000 to 5600cm-1and prominent bands of CaAl-LDH-Cl intercalated with SDS around 8388cm-1. This band was assigned to the second overtone of the first fundamental of CH stretching vibrations of SDS, and it could be used to determinate the result of CaAl-LDH-Cl modified by SDS. Moreover, the results revealed that different adsorption behaviors were observed at different (high and low) concentrations of SDS. When the SDS concentration was around 0.2molL-1, anion exchange intercalation occurred and the interlayer distance expanded to about 3.25nm. When SDS concentration was 0.005molL-1, the surface adsorption of DS- was the major anion exchange event.

Eating for health and the environment: Australian regulatory responses for dietary change

The Australian food system significantly contributes to a range of key environmental issues including harmful greenhouse gas emissions, air pollution, soil desertification, biodiversity loss and water scarcity. At the same time, the Australian s food system is a key cause of public health nutrition issues that stem from the co-existence of over- and under-consumption of dietary energy and nutrients. Within these challenges lie synergies and opportunities because a diet that has a lower environmental impact generally aligns with good nutrition. Australian State and Federal initiatives to influence food consumption patterns focus on individual body weight and ‘soft law’ interventions. These regulatory approaches, by focusing on select symptoms of food system failures, are fragmented, reductionist and inefficient. In order to illustrate this point, this paper will explore Australian regulatory responses to diet-related illnesses. The analysis will support the argument that only when regulatory responses to diets become embedded within reform of the current food system will substantial improvements to human and planetary health be achieved.

International regulatory framework for food production and diversity

Stakeholders commonly agree that food systems need to be urgently reformed. Yet, how food systems should be reformed is extremely contested. Public international law and regulations are uniquely placed to influence and guide law, policy, programmes and action at regional, national and local levels. Although plenty of international legal instruments intersect with food-related issues, the international regulation of food systems is fragmented, understudied and contested. In order to address these issues, this paper maps and analyses the public international regulatory aspects of food production with a view to providing recommendations for reform. Accordingly, this paper brings together a variety of binding and non-binding international regulatory instruments that to varying degrees and from a range of angles deals with the first activity in the food system: food production. The following paper traces the regulatory tools from natural resources, to the farmers and farm workers that apply skill and experience, and finally to the different dimension of world trade in food. The various regulatory instruments identified, and their collective whole, will be analysed against a rights-based approach to food security.

The role of regulatory frameworks and non-state actors in creating sustainable diets

This presentation discussed the growing recognition of sustainable diets at international governance levels and how this reflects the challenges and win-win opportunities of living within our ecological limits. I assert that sustainable diets provide an example of how living within our ecological limits would actually make us better off even apart from environmental benefits. After determining whether Australians’ generally have a sustainable diet, I outlined how Australian regulators are attempting to address sustainable diets. I argued that the personal responsibility approach coupled with the focus on preventing or reducing overweight and obesity levels are proving incapable of bringing about long-term sustainable diets that will contribute to the health and well-being of Australian people.

Gene regulation by translational inhibition is determined by Dicer partnering proteins

MicroRNAs (miRNAs) are small regulatory RNAs produced by Dicer proteins that regulate gene expression in development and adaptive responses to the environment1,​2,​3,​4. In animals, the degree of base pairing between a miRNA and its target messenger RNA seems to determine whether the regulation occurs through cleavage or translation inhibition1. In contrast, the selection of regulatory mechanisms is independent of the degree of mismatch between a plant miRNA and its target transcript5. However, the components and mechanism(s) that determine whether a plant miRNA ultimately regulates its targets by guiding cleavage or translational inhibition are unknown6. Here we show that the form of regulatory action directed by a plant miRNA is determined by DRB2, a DICER-LIKE1 (DCL1) partnering protein. The dependence of DCL1 on DRB1 for miRNA biogenesis is well characterized7,​8,​9, but we show that it is only required for miRNA-guided transcript cleavage. We found that DRB2 determines miRNA-guided translational inhibition and represses DRB1 expression, thereby allowing the active selection of miRNA regulatory action. Furthermore, our results reveal that the core silencing proteins ARGONAUTE1 (AGO1) and SERRATE (SE) are highly regulated by miRNA-guided translational inhibition. DRB2 has been remarkably conserved throughout plant evolution, raising the possibility that translational repression is the ancient form of miRNA-directed gene regulation in plants, and that Dicer partnering proteins, such as human TRBP, might play a similar role in other eukaryotic systems.